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Cited by 2Cholesterol and Lipid MetabolismNeuroinflammation and Neurodegeneration MechanismsNeuroscience and Neuropharmacology Research

The DOI matches its Crossref record; 100 of the 100 references it cites were found in a registry or an index; 2 of the 108 sources its claims cite were checked for support.

LDL Cholesterol Modulates Astrocyte Metabolism, Lipid Handling, and Morphology: Evidence From In Vitro and In Vivo Models

Open access
Baumart, Gabriela Joras, Rodrigues, Matheus Scarpatto, Sholl, Juliete Nathali, de Sousa, Arieli Cruz, Weber, Augusto Ferreira, Costa‐Beber, Lílian Corrêa, Farias, Hémelin Resende, Costa, Mariana Viana, Peres, Ariadni Mesquita, de Camargo, Pedro Rocha, Fróes, Fernanda Telles, Bast, Rachel Krolow, de Bem, Andreza Fabro, Figueiró, Fabrício, Guma, Fátima T. C. R., de Oliveira, Jade·2026·Journal of Neurochemistry
DOI 10.1111/jnc.70403PMC PMC12982915
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Abstract

Astrocytes are the primary antioxidant defense cells of the brain, protecting the central nervous system (CNS) through a controlled inflammatory response and acting as metabolic suppliers to neurons. These cells exhibit morphological, functional, and molecular changes in pathological conditions, such as neurodegenerative diseases. Previous studies have demonstrated a link between hypercholesterolemia, especially elevated levels of low‐density lipoprotein (LDL) cholesterol, and brain disorders, including hippocampal astrogliosis. In this context, this study aimed to investigate how LDL cholesterol modulates astrocyte biology. In vitro, high‐passage rat C6 astroglial cells were exposed to human LDL cholesterol (50 or 300 μg/mL) for 24 or 48 h. We evaluated lipid accumulation, cholesterol metabolism‐related gene expression, astrocyte‐related gene expression, reactive species production, antioxidant activity, redox‐related gene expression, fatty acid and glucose uptake, cell proliferation, and metabolic activity ‐ 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) reduction. LDL exposure increased intracellular lipid content and downregulated LDL receptor (LDLR), 3‐hydroxy‐3‐methylglutaryl‐coenzyme A reductase (HMGCR), and sterol regulatory element‐binding transcription factor 1 (SREBF1) gene expression. LDL exposure altered astrocytic marker expression, as evidenced by increased glial fibrillary acid protein (GFAP) messenger RNA (mRNA) levels at 24 h with 300 μg/mL LDL and at 48 h with 50 μg/mL LDL. LDL cholesterol decreased long‐chain fatty acids (LCFA) uptake and superoxide dismutase (SOD) activity at 24 h and increased cluster of differentiation 36 (CD36), also known as fatty acid translocase levels, at 48 h. Nuclear factor erythroid 2‐related factor 2 (NRF2) expression was significantly increased after 48 h of incubation with 300 μg/mL LDL. The MTT reduction assay did not indicate decreased cell viability; instead, it revealed increased metabolic activity after 24 h of incubation with 300 μg/mL LDL, with no changes observed in glucose uptake. In vivo, hippocampal astrocytes from young (3‐month‐old) and middle‐aged (14‐month‐old) LDL receptor knockout (LDLr−/−) and wild‐type C57BL/6 mice were analyzed by immunofluorescence and quantitative reverse transcription polymerase chain reaction (RT‐qPCR). In the hippocampal Cornu Ammonis 3 (CA3) region, 14‐month‐old LDLr−/− mice showed an increase in the number of processes compared to 3‐month‐old wild‐type C57BL/6 mice. Aging and genotype influenced astrocyte morphology and expression of genes such as S100 calcium‐binding protein B (S100B) and aquaporin‐4 (AQP4). Our findings demonstrate that LDL cholesterol induces morphological, metabolic, and molecular changes in astrocytes, both in vitro and in vivo, suggesting that astroglial cells are sensitive to lipid imbalance and may play a role in the brain consequences of hypercholesterolemia. image

Overview

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Written from the paper’s full text.

TL;DR

LDL cholesterol exposure promotes lipid droplet accumulation, suppresses cholesterol synthesis/uptake genes, and alters astrocyte markers and metabolism in C6 astroglial cells. In LDLr−/− mice, aging and genotype produce region-specific hippocampal astrocyte morphological and gene-expression changes, suggesting astrocytes are sensitive to lipid imbalance.

Key findings

  • In C6 cells, 300 μg/mL LDL increased intracellular lipid accumulation at 24 h (Ctrl vs. 300 μg/mL, p < 0.0001) and 48 h (p = 0.0354), whereas 50 μg/mL did not. LDL also downregulated LDLR (300 μg/mL: 24 h p = 0.0013; 48 h p = 0.0343), HMGCR (300 μg/mL: 24 h p = 0.0005; 48 h p = 0.0004), and SREBF1 (24 h, 300 μg/mL p = 0.0099).
  • GFAP mRNA increased at 24 h with 300 μg/mL LDL (p < 0.0001) and at 48 h with 50 μg/mL LDL (p = 0.0275). MTT reduction increased at 24 h with 300 μg/mL LDL (p < 0.0001); LCFA uptake decreased at 24 h with 300 μg/mL LDL (p = 0.0090); CD36 increased at 48 h with 300 μg/mL LDL (p = 0.0175); SOD activity decreased at 24 h with 50 μg/mL LDL (p = 0.0278); NRF2 mRNA increased at 48 h with 300 μg/mL LDL (p = 0.0020). Glucose uptake, proliferation, and cell death showed no significant changes.
  • In vivo, 14-month-old LDLr−/− mice had more astrocytic processes in hippocampal CA3 than 3-month-old wild-type mice (p = 0.0233) and 3-month-old LDLr−/− mice (p = 0.0399). S100B showed an age effect (p = 0.02464), with increased expression in 14- vs. 3-month wild-type mice (p = 0.009154); AQP4 showed a robust age effect (p = 0.00003), with reduced expression at 14 months in both genotypes (wild-type p = 0.000305; LDLr−/− p = 0.004994) and lower expression in 3-month LDLr−/− vs. wild-type mice (p = 0.046970).

Methods

  • In vitro: high-passage rat C6 astroglial cells were exposed to human LDL cholesterol (50 or 300 μg/mL) or PBS for 24 or 48 h. Outcomes included lipid accumulation, lysosomal content, CD36, LCFA and glucose uptake, RT-qPCR, MTT reduction, reactive species, SOD activity, SRB, Ki-67, and Annexin V/PI.
  • In vivo: male C57BL/6 wild-type and LDLr−/− mice at 3 and 14 months (3–5 animals per group) were used. Hippocampal astrocytes were analyzed by GFAP immunofluorescence with Simple Neurite Tracer/Sholl analysis and by RT-qPCR for astrocytic genes.
  • Statistics: normality testing followed by one-way ANOVA with Dunnett’s or Kruskal–Wallis with Dunn’s post hoc tests in vitro; two-way ANOVA with Duncan’s post hoc test in vivo. Significance was p < 0.05, with p ≤ 0.1 considered a statistical tendency.

Limitations

  • The paper states that the high-passage C6 glioma cell line may not fully recapitulate primary astrocytes, that acute exposure may miss chronic effects, and that the in vivo sample size was small (n = 3–5) with no formal sample-size calculation. It also states that LDL concentrations in the CNS under BBB disruption are unknown, that no direct measurement of LDL entry into the brain was performed, that no randomization was used, and that residual blood components may influence qPCR analyses.
  • The text makes evident additional constraints: some in vitro RT-qPCR experiments used n = 2–4, post hoc sensitivity for pairwise comparisons with Cohen’s d = 0.8 was approximately 0.16, in vivo analyses used no multiple-comparison adjustments, and only male mice were studied.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1111/jnc.70403

Cites

100 of 100 references verified0 citation errors · 0 not checkable

Statistics

Verification

Claims the paper makes

2 of 108 cited sources checked · 0 supported · 0 flagged · accuracy not yet measured

  • Premisecitation

    Treatment with lipolysis products increases lipid droplet accumulation in cultured astrocytes.

    “For example, treatment with lipolysis products has been shown to increase lipid droplet (LD) accumulation, upregulate inflammation‐related gene expression, elevate cellular stress responses, and enhance cytokine release (Liu et al. 2015; Lee et al. 2017).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites Liu et al. 2015 source

      The Results sections on mitochondrial mutants and neuronal mitochondrial dysfunction report lipid-droplet accumulation in fly glia, not cultured astrocytes treated with lipolysis products. The section on lipid peroxidation reports that lipase overexpression reduces droplet formation. Searches for lipolysis products and astrocyte treatment found no passage addressing the claimed treatment. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites Lee et al. 2017 source
  • Premisecitation

    Incubation with LDL alters astrocyte morphology.

    “Moreover, incubation with LDL has been shown to alter astrocyte morphology, although the specific consequences and mechanisms by which LDL cholesterol affects astrocytes remain incompletely understood (Dehouck et al. 1994; Engel et al. 2019; Pitas et al. 1987).”
    Quote in the paperSubstantial claim1 of 3 sources checked
    • cites Dehouck et al. 1994 source

      The section “Cholesterol Content and Binding of LDL on Astrocytes” reports changes in LDL binding and cholesterol content after incubation in lipoprotein-deficient serum, not changes in astrocyte morphology after incubation with LDL. Searches for astrocyte morphology, shape, and LDL incubation found no passage addressing the claimed morphological effect. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites Engel et al. 2019 source
    • cites Pitas et al. 1987 source
  • Findingown data

    In C6 cells, treatment with 50 μg/mL LDL significantly upregulated GFAP gene expression at 48 h.

    “while a significant upregulation was observed at 48 h in cells exposed to 50 μg/mL LDL (F(2, 9) = 4.874, p = 0.0413, Dunnett's post hoc test: Ctrl vs. 50 μg/mL: *p = 0.0275; Ctrl vs. 300 μg/mL: p = 0.5513) (Figure 3c).”
    Quote in the paperSubstantial claim
  • Findingown data

    In C6 astroglial cultures, exposure to 300 μg/mL LDL cholesterol for 24 or 48 h reduced LDLR mRNA expression compared with control.

    “Using RT‐qPCR, we observed a reduction in LDLR expression after exposure to 300 μg/mL LDL for 24 or 48 h (Figure 2a)”
    • Also stated in: § Incubation With LDL Cholesterol Decreases Gene Expression of LDLR, 3‐Hydroxy‐3‐Methylglutaryl‐Coenzyme A Reductase (HMGCR), and Sterol Regulatory Element‐Binding Transcription Factor 1 (SREBF1) · plain
      “LDLr messenger RNA (mRNA) and (b) HMGCR mRNA are downregulated at 300 μg/mL LDL cholesterol at both incubation times”
    Quote in the paperSubstantial claim
  • Findingown data

    LDL cholesterol decreases gene expression of LDLR in C6 cells.

    “Incubation With LDL Cholesterol Decreases Gene Expression of LDLR, 3‐Hydroxy‐3‐Methylglutaryl‐Coenzyme A Reductase (HMGCR), and Sterol Regulatory Element‐Binding Transcription Factor 1 (SREBF1)”
    • Also stated in: § Abstract · plain
      “LDL exposure increased intracellular lipid content and downregulated LDL receptor (LDLR), 3‐hydroxy‐3‐methylglutaryl‐coenzyme A reductase (HMGCR), and sterol regulatory element‐binding transcription factor 1 (SREBF1) gene expression.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The C6 rat astroglial cell line is a valid model for studying astrocyte biology.
    • Rests on an assumption: The LDL cholesterol concentrations used in vitro (50 and 300 μg/mL) are physiologically relevant to the brain.
    • Rests on an assumption: The isolated human LDL cholesterol used in vitro is not oxidized and is biologically representative of native LDL.
    • Rests on an assumption: Changes in mRNA levels of astrocytic genes reflect corresponding changes in protein levels or functional astrocyte states.
  • It may state two separate claims as one. 39
  • The quote only partly states this claim. 1
  • Probably not a substantial claim. 51

  • It may state two separate claims as one.

    Findingown data

    In C6 astroglial cells, after 24 h, exposure to 300 μg/mL LDL cholesterol significantly reduced external long-chain fatty acid uptake, whereas no significant differences were observed at 50 μg/mL.

    “After 24 h, cells exposed to 300 μg/mL LDL cholesterol showed a significant reduction in external fatty‐acid uptake (F(2, 6) = 9.537, p = 0.0137, Dunnett's post hoc test: Ctrl vs. 300 μg/mL: **p = 0.0090) (Figure 4c,d; Figure S1f), whereas no significant differences were observed at 50 μg/mL (F(2, 6) = 9.537, p = 0.0137, Dunnett's post hoc test: Ctrl vs. 50 μg/mL: p = 0.2923).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    In C57BL/6 wild-type mice, S100B expression was upregulated with age (3 vs. 14 months; p=0.009154), whereas no significant age-related change was observed in LDLr−/− mice.

    “Duncan's post hoc analysis demonstrated an age‐dependent upregulation of S100B expression in C57BL/6 wild‐type mice (3 vs. 14 months; **p = 0.009154), whereas no significant age‐related change was observed in LDLr−/− mice.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    In C6 astroglial cultures, exposure to 300 μg/mL LDL cholesterol for 24 h reduced SREBF1 mRNA levels compared with control, but not at 48 h.

    “as well as in SREBF1 mRNA levels after 24 h of incubation with 300 μg/mL LDL, the gene that encodes the transcription factor SREBP‐1 (Figure 2c) (24 h, H(2) = 8.018, p = 0.0014, Dunn's post hoc test: Ctrl vs. 50 μg/mL: p = 0.2603; Ctrl vs. 300 μg/mL: **p = 0.0099), (48 h, H(2) = 2.636, p = 0.2899, Dunn's post hoc test: Ctrl vs. 50 μg/mL: p = 0.5728; Ctrl vs. 300 μg/mL: p = 0.2286)”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    Aging and genotype influenced astrocyte morphology and expression of genes such as S100B and AQP4.

    “Aging and genotype influenced astrocyte morphology and expression of genes such as S100 calcium‐binding protein B (S100B) and aquaporin‐4 (AQP4).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: LDL cholesterol reaches astrocytes in the brain in the in vivo hypercholesterolemia models used.
    • Rests on an assumption: The LDLr−/− mouse model used here develops brain exposure to LDL cholesterol relevant to hypercholesterolemia.
    • Rests on an assumption: Changes in mRNA levels of astrocytic genes reflect corresponding changes in protein levels or functional astrocyte states.

  • It may state two separate claims as one.

    Findingown data

    In C6 cells, incubation with 300 μg/mL LDL cholesterol increased metabolic activity after 24 h, but not after 48 h.

    “Incubation with 300 μg/mL LDL cholesterol increased metabolic activity after 24 h (F(2, 6) = 142.0, p < 0.0001, Dunnett's post hoc test: Ctrl vs. 50 μg/mL: p = 0.1004; Ctrl vs. 300 μg/mL: ****p < 0.0001), while the other incubation time did not show significant differences in metabolic activity (48 h, H(2) = 7.200, p = 0.0036, Dunn's post hoc test: Ctrl vs. 50 μg/mL: p = 0.3594; Ctrl vs. 300 μg/mL”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    In C6 cells, exposure to 300 μg/mL LDL showed a trend toward increased NRF2 gene expression after 24 h, which became statistically significant after 48 h.

    “C6 cells exposed to 300 μg/mL LDL showed a trend toward increased NRF2 gene expression after 24 h (F(2, 9) = 3.253, p = 0.0865, Dunnett's post hoc test: Ctrl vs. 50 μg/mL: p = 0.2514; Ctrl vs. 300 μg/mL: #p = 0.0576), which became statistically significant after 48 h of incubation (F(2, 8) = 15.18, p = 0.0019, Dunnett's post hoc test: Ctrl vs. 50 μg/mL: p = 0.9581; Ctrl vs. 300 μg/mL: **p = 0.0020”
    Quote in the paperNot a substantial claim

  • The quote only partly states this claim.

    Findingown data

    In C6 astrocytes, LDL cholesterol exposure caused intracellular accumulation of lipid species.

    “Importantly, astrocytes presented intracellular accumulation of lipid species after being challenged with LDL cholesterol.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    At 3 months of age, LDLr−/− mice exhibited reduced hippocampal AQP4 expression compared with age-matched C57BL/6 mice, a difference no longer evident at 14 months.

    “In addition, LDLr−/− mice exhibited reduced hippocampal AQP4 expression compared with age‐matched C57BL/6 mice at 3 months of age (**p = 0.046970), a difference that was no longer evident at 14 months of age.”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    In C6 cells, 300 μg/mL LDL cholesterol increased CD36 content at 48 h of incubation, while no significant change was observed at 50 μg/mL in the same period.

    “On the other hand, 300 μg/mL LDL cholesterol increased the CD36 content at 48 h of incubation, while no significant change was observed at 50 μg/mL in the same incubation period (F(2, 6) = 6.956, p = 0.0274, Dunnett's post hoc test: Ctrl vs. 50 μg/mL: p = 0.1340; Ctrl vs. 300 μg/mL: *p = 0.0175]”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    In the present study, astrocytic morphology is altered by age and is region specific: more pronounced structural alterations, such as increased process number, appear later and are preferentially localized to the CA3 hippocampal region.

    “Importantly, we show that astrocytic morphology is altered by age and is region specific: While biochemical markers of activation emerge early, more pronounced structural alterations, such as increased process number, appear later and are preferentially localized to the CA3 hippocampal region.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    In 2024, high levels of blood LDL cholesterol were officially recognized as a strong midlife risk factor for dementia.

    “In fact, in 2024, high levels of blood LDL cholesterol were officially recognized as a strong midlife risk factor for dementia (Livingston et al. 2024).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Livingston et al. 2024 source

  • It may state two separate claims as one.

    Premisecitation

    Astrogliosis, classically characterized by increased GFAP immunoreactivity, is a prominent feature of hypercholesterolemic animal models and frequently co-localizes with regions of BBB disruption.

    “Consistent with this role, astrogliosis, classically characterized by increased glial fibrillary acid protein (GFAP) immunoreactivity, is a prominent feature of hypercholesterolemic animal models and frequently co‐localizes with regions of BBB disruption (Ramírez et al. 2011; de Oliveira et al. 2014; de Oliveira, Engel, de Paula, Dos Santos, et al. 2020).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Ramírez et al. 2011 source
    • cites de Oliveira et al. 2014 source
    • cites de Oliveira, Engel, de Paula, Dos Santos, et al. 2020

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Hypercholesterolemia is a well-established risk factor for cardiovascular disease.

    “Hypercholesterolemia, defined by elevated plasma low‐density lipoprotein (LDL) cholesterol, is a well‐established risk factor not only for cardiovascular disease (Borén et al. 2022; Steinberg 2013; Tabas et al. 2015; Boffa and Koschinsky 2019), but also for cognitive decline and dementia in later life (Kivipelto et al. 2001; Kivipelto and Solomon 2006; Livingston et al. 2024; Sparks et al. 2000, 2”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Borén et al. 2022 source
    • cites Steinberg 2013 source
    • cites Tabas et al. 2015 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Hypercholesterolemia is a well-established risk factor for cognitive decline and dementia in later life.

    “Hypercholesterolemia, defined by elevated plasma low‐density lipoprotein (LDL) cholesterol, is a well‐established risk factor not only for cardiovascular disease (Borén et al. 2022; Steinberg 2013; Tabas et al. 2015; Boffa and Koschinsky 2019), but also for cognitive decline and dementia in later life (Kivipelto et al. 2001; Kivipelto and Solomon 2006; Livingston et al. 2024; Sparks et al. 2000, 2”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Kivipelto et al. 2001 source
    • cites Kivipelto and Solomon 2006 source
    • cites Livingston et al. 2024 source

  • It may state two separate claims as one.

    Premisecitation

    LDs may act as indirect markers of oxidative stress, as oxidized lipids are transferred from neurons to astrocytes through neuron-astrocyte metabolic coupling mechanisms.

    “Emerging evidence further suggests that LDs may act as indirect markers of oxidative stress, as oxidized lipids are transferred from neurons to astrocytes through neuron–astrocyte metabolic coupling mechanisms (Liu and Zhang 2014; Smolič et al. 2021).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Liu and Zhang 2014 source
    • cites Smolič et al. 2021 source

  • Probably not a substantial claim. The authors may not put it forward as true. The paper’s conclusions may not depend on it.

    Interpretationown data

    The reduction in SOD activity observed after 24 h of exposure may be related to an early oxidative stress event.

    “Accordingly, we hypothesize that the reduction in SOD activity observed after 24 h of exposure may be related to an early oxidative stress event.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim.

    Study design and conductreasoning

    In our study, the cells were used for a maximum of five passages after thawing to minimize phenotypic drift.

    “To minimize this risk, in our study, the cells were used for a maximum of five passages after thawing.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim.

    Interpretationreasoning

    At 24 h, astrocyte cultures do not exhibit detectable increases in reactive species due to the action of other antioxidant mechanisms independent of SOD activity.

    “We further propose that, at this time point, astrocyte cultures do not exhibit detectable increases in reactive species due to the action of other antioxidant mechanisms independent of SOD activity.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductnone given

    Due to practical constraints, the final sample size consisted of 3-5 animals per group.

    “Due to practical constraints, the final sample size consisted of 3–5 animals per group.”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Interpretationcitation

    The simultaneous decrease in SREBF1 suggests a deeper disruption in lipid regulatory pathways, and sustained LDL exposure may impair the astrocytic regulatory machinery.

    “On the other hand, the simultaneous decrease in SREBF1, a master regulator of lipid metabolism, suggests a deeper disruption in lipid regulatory pathways. Sustained LDL exposure may impair the astrocytic regulatory machinery itself, potentially compromising critical functions such as cholesterol delivery to neurons, membrane remodeling, and metabolic adaptability (Shimano and Sato 2017).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Shimano and Sato 2017 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. The paper may give no reason why it holds.

    Study design and conductnone given

    The C6 cell line is not listed in the International Cell Line Authentication Committee (ICLAC) Register of Misidentified Cell Lines.

    “The C6 cell line is not listed in the International Cell Line Authentication Committee (ICLAC) Register of Misidentified Cell Lines.”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Interpretationcitation

    The APOE mRNA trend toward decrease after LDL exposure contrasts with in vivo studies where hypercholesterolemia increased ApoE protein levels in rabbit cortex, underscoring differences between in vitro and in vivo systems.

    “In this context, our observation that APOE mRNA expression also tended to decrease is noteworthy, although it contrasts with in vivo studies in which hypercholesterolemia increased ApoE protein levels in rabbit cortex (Sparks et al. 1995), underscoring key differences between simplified in vitro systems and complex in vivo environments.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Sparks et al. 1995 source

  • It may state two separate claims as one.

    Interpretationcitation

    The temporal pattern of NRF2 and SOD reflects activation of the Nrf2-ARE axis, which promotes transcription of antioxidant enzymes and restores redox balance.

    “We propose that this temporal pattern reflects activation of the nuclear factor erythroid 2‐related factor 2‐antioxidant response element (Nrf2‐ARE) axis, which promotes the transcription of antioxidant enzymes and restores redox balance (Sigfridsson et al. 2018; He and Hewett 2025).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Sigfridsson et al. 2018 source
    • cites He and Hewett 2025 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductnone given

    The study was approved by the Ethics Committee at UFRGS (Ethics Committee Approved Project Number 43754).

    “This project was approved by the Ethics Committee at UFRGS (Ethics Committee Approved Project Number 43754).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Sparks et al. (1995) showed that amyloid plaques were formed when rabbits were fed a hypercholesterolemic diet.

    “Sparks et al. (1995) showed that amyloid plaques were formed when rabbits were fed a hypercholesterolemic diet.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Sparks et al. (1995) source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductnone given

    All animal procedures were performed following the local ethics committee and the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health.

    “All animal procedures were performed following the local ethics committee and the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health (Bethesda, MD, USA).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Interpretationreasoning

    In the complex in vivo environment, multiple cell types experience chronic hypercholesterolemia-induced stress, creating sustained demand for antioxidant defenses that eventually become overwhelmed, whereas the controlled in vitro system allows astrocytes to mount an effective compensatory response.

    “In the complex in vivo environment, multiple cell types experience chronic hypercholesterolemia‐induced stress, creating sustained demand for antioxidant defenses that eventually become overwhelmed. In contrast, our controlled in vitro system allows astrocytes to mount an effective compensatory response through their intrinsic antioxidant machinery.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Interpretationown data

    In our experimental model, exposure to LDL cholesterol did not alter the proliferation levels of the astrocyte culture, confirming other studies showing that the reactive astrocytic phenotype does not always promote proliferation.

    “In our experimental model, exposure to LDL cholesterol did not alter the proliferation levels of the astrocyte culture, confirming other studies showing that the reactive astrocytic phenotype does not always promote proliferation (Liddelow and Barres 2017).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Liddelow and Barres 2017 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Study design and conductnone given

    Outliers were defined, and then, excluded after analysis.

    “Outliers were defined, and then, excluded after analysis.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper may give no reason why it holds.

    Interpretationnone given

    BBB dysfunction may permit the entry of LDL cholesterol into the CNS under peripheral inflammatory conditions, such as hypercholesterolemia and metabolic disease.

    “Therefore, BBB dysfunction may permit the entry of LDL cholesterol into the CNS under peripheral inflammatory conditions, such as hypercholesterolemia and metabolic disease.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductown data

    The in vivo data were analyzed by two-way ANOVA followed by Duncan's post hoc test, without multiple comparison adjustments.

    “The in vivo data are expressed as mean ± standard deviation (SD) and were analyzed by two‐way ANOVA followed by using Duncan's post hoc test, without multiple comparison adjustments.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductnone given

    No formal calculations were conducted to determine the sample size for any of the animal procedures.

    “No formal calculations were conducted to determine the sample size for any of the animal procedures.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper may give no reason why it holds. It may state two separate claims as one.

    Interpretationnone given

    Once within the CNS, LDL cholesterol can exert neurotoxic effects on neurons and disrupt glial function.

    “Once within the CNS, LDL cholesterol can exert neurotoxic effects on neurons and disrupt glial function.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    LDL exposure induces mitochondrial dysfunction in primary cortical neurons and neuronal cell lines.

    “In parallel, studies examining the effects of native (non‐oxidized) and oxidized LDL (ox‐LDL) on brain cells have reported that LDL exposure in primary cortical neurons and neuronal cell lines disrupts endolysosomal function, increases amyloid‐β production, reduces synaptophysin levels, and induces mitochondrial dysfunction and oxidative stress (Engel et al. 2016; Hui et al. 2012; Yamchuen et al.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Engel et al. 2016 source
    • cites Hui et al. 2012 source
    • cites Yamchuen et al. 2017 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductreasoning

    The study was initially designed to detect medium-to-large main effects (Cohen's f = 0.35-0.40) with α = 0.05 and 1−β = 0.80, which indicated a target of approximately six animals per Genotype × Age condition.

    “Therefore, the study was initially designed to detect medium‐to‐large main effects (Cohen's f = 0.35‐0.40) with α = 0.05 and 1−β = 0.80, which indicated a target of approximately six animals per Genotype × Age condition.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. The paper may give no reason why it holds. It may state two separate claims as one.

    Interpretationreasoning

    Decreased glucose uptake may be related to the metabolic state of the cells, and such a decrease should reduce lactate production, an essential nutrient for neurons.

    “This potential alteration in glucose uptake should be further investigated, as decreased glucose uptake may be related to the metabolic state of the cells, because this decrease should also reduce the production of lactate, an essential nutrient for neurons.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. The paper may give no reason why it holds. It may state two separate claims as one.

    Study design and conductnone given

    Residual blood components may still be present in the brain tissue and influence the qPCR analyses.

    “Residual blood components may still be present in the brain tissue and influence the qPCR analyses.”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Interpretationreasoning

    In summary, astrocytes exhibit some resistance to oxidative stress; after a short incubation with LDL cholesterol, ROS are generated, which does not persist with longer incubation times, likely due to antioxidant mechanisms.

    “In summary, astrocytes exhibit some resistance to oxidative stress. After a short incubation period with LDL cholesterol, there is the generation of ROS, which does not persist with longer incubation times, likely due to the action of antioxidant mechanisms.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim.

    Premisecitation

    The specific consequences and mechanisms by which LDL cholesterol affects astrocytes remain incompletely understood.

    “Moreover, incubation with LDL has been shown to alter astrocyte morphology, although the specific consequences and mechanisms by which LDL cholesterol affects astrocytes remain incompletely understood (Dehouck et al. 1994; Engel et al. 2019; Pitas et al. 1987).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites Dehouck et al. 1994 source
    • cites Engel et al. 2019 source
    • cites Pitas et al. 1987 source

  • It may state two separate claims as one.

    Interpretationown data

    LDL cholesterol appears to elicit a transient oxidative stress response in astrocytes that is followed by activation of protective mechanisms.

    “LDL cholesterol appears to elicit a transient oxidative stress response in astrocytes that is followed by activation of protective mechanisms.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Interpretationreasoning

    Astrocytes possess robust protective capabilities that can respond to acute LDL exposure, but these same mechanisms become insufficient under chronic pathological conditions.

    “This demonstrates that astrocytes possess robust protective capabilities that can respond to acute LDL exposure, but these same mechanisms become insufficient under chronic pathological conditions.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductnone given

    No randomization was performed to allocate animals in the study.

    “No randomization was performed to allocate animals in the study.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    LDL exposure disrupts endolysosomal function in primary cortical neurons and neuronal cell lines.

    “In parallel, studies examining the effects of native (non‐oxidized) and oxidized LDL (ox‐LDL) on brain cells have reported that LDL exposure in primary cortical neurons and neuronal cell lines disrupts endolysosomal function, increases amyloid‐β production, reduces synaptophysin levels, and induces mitochondrial dysfunction and oxidative stress (Engel et al. 2016; Hui et al. 2012; Yamchuen et al.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Engel et al. 2016 source
    • cites Hui et al. 2012 source
    • cites Yamchuen et al. 2017 source

  • It may state two separate claims as one.

    Interpretationcitation

    Reduced LDLR expression may limit cholesterol overload but could also impair processes beyond lipid handling, and because LDLR contributes to Aβ clearance, its persistent downregulation could favor Aβ accumulation.

    “For example, reduced LDLR expression may limit cholesterol overload but could also impair processes beyond lipid handling. Because LDLR contributes to Aβ clearance in the brain (Basak et al. 2012), its persistent downregulation could favor Aβ accumulation, a hallmark of neurodegenerative pathology.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Basak et al. 2012 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductown data

    Post hoc evaluation of the achieved sensitivity indicated that pairwise comparisons with expected large effect sizes (Cohen's d = 0.8) corresponded to an average detectable power of approximately 0.16.

    “Post hoc evaluation of the achieved sensitivity indicated that pairwise comparisons with expected large effect sizes (Cohen's d = 0.8) corresponded to an average detectable power of approximately 0.16.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Study design and conductreasoning

    No animals were excluded, as all met the established criteria and remained in good condition throughout the study.

    “No animals were excluded, as all met the established criteria and remained in good condition throughout the study.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductreasoning

    The absence of direct measurements of LDL penetration into the hypercholesterolemic brain represents an important limitation.

    “Nonetheless, the absence of direct measurements of LDL penetration into the hypercholesterolemic brain represents an important limitation.”
    Quote in the paperNot a substantial claim
    • The authors state this as an assumption: Physiological LDL concentrations in the brains of humans or mice with hypercholesterolemia are unknown, because LDL is not normally found in the CNS and its concentration under pathological BBB disruption has not been quantified.

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    The MTT reagent can cross cell and mitochondrial membranes, and its intracellular reduction is mainly mediated by oxidoreductase, dehydrogenase, and electron donor enzymes.

    “The MTT reagent can cross cell and mitochondrial membranes, and its intracellular reduction is mainly mediated by oxidoreductase, dehydrogenase, and electron donor enzymes (Ghasemi et al. 2021).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Ghasemi et al. 2021 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    We previously observed an increased immunoreactivity of AQP4 in the hippocampus of LDLr−/− mice.

    “We previously observed an increased immunoreactivity of this protein in the hippocampus of LDLr−/− mice (de Oliveira et al. 2014).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites de Oliveira et al. 2014 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductown data

    The in vivo findings should be interpreted with caution due to the small sample size.

    “However, these in vivo findings should be interpreted with caution due to the small sample size.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim.

    Premisecitation

    LCFA and lipoproteins such as LDL and ox-LDL are ligands for CD36.

    “In addition, LCFA and lipoproteins such as LDL and ox‐LDL are ligands for CD36 (Park 2014).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Park 2014 source

  • Probably not a substantial claim.

    Premisecitation

    Astrocytes are essential CNS cells for glucose uptake.

    “Astrocytes are essential CNS cells for glucose uptake (Pellerin and Magistretti 1994; Pellerin 2018).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Pellerin and Magistretti 1994 source
    • cites Pellerin 2018 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Previous studies have reported increased astrocyte density in the hippocampus of LDLr−/− mice.

    “Previous studies have reported increased astrocyte density in the hippocampus of LDLr−/− mice (de Oliveira et al. 2014; de Oliveira, Engel, de Paula, Dos Santos, et al. 2020) and have identified age‐dependent heterogeneity in astrocytic reactivity profiles (Clarke et al. 2018; Lee et al. 2022).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites de Oliveira et al. 2014 source
    • cites de Oliveira, Engel, de Paula, Dos Santos, et al. 2020

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • It may state two separate claims as one.

    Interpretationown data

    These changes are consistent with astrocytic hypertrophy and suggest a reactive phenotype associated with disrupted lipid metabolism.

    “These changes are consistent with astrocytic hypertrophy and suggest a reactive phenotype associated with disrupted lipid metabolism (Owasil et al. 2020; Dai et al. 2023).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Owasil et al. 2020 source
    • cites Dai et al. 2023 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Caspase 3 and GFAP colocalization was not verified in these animals.

    “However, caspase 3 and GFAP colocalization was not verified in these animals (de Oliveira, Engel, de Paula, Melo, et al. 2020).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites de Oliveira, Engel, de Paula, Melo, et al. 2020

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    Studies have indicated that the levels of expression of AQP4 increased in mouse models of AD and in humans with the disease, a situation that may favor the reported disturbances in water homeostasis and increased BBB permeability.

    “Studies have indicated that the levels of expression of AQP4 increased in mouse models of AD and in humans with the disease, a situation that may favor the reported disturbances in water homeostasis and increased BBB permeability (Wilcock et al. 2009).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Wilcock et al. 2009 source

  • It may state two separate claims as one.

    Premisecitation

    Fourteen months represents middle age in mice, corresponding to a period of increased cognitive vulnerability in patients with FH, whereas 3 months reflects an early stage of pathology.

    “Fourteen months represents middle age in mice (Dutta and Sengupta 2016), corresponding to a period of increased cognitive vulnerability in patients with FH, whereas 3 months reflects an early stage of pathology.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Dutta and Sengupta 2016 source

  • Probably not a substantial claim. It may be too vague to check against evidence.

    Interpretationreasoning

    The findings have potential implications for neurodegenerative disease risk and progression.

    “Taken together, our findings identify astrocytes as key cellular mediators of cholesterol‐induced brain dysfunction and provide new mechanistic insight into how peripheral hypercholesterolemia can disrupt central nervous system homeostasis, with potential implications for neurodegenerative disease risk and progression.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    In aging, S100B expression showed regional specificity; however, the functional consequences of these specificity alterations remained unknown.

    “In aging, the S100B expression showed regional specificity; however, the functional consequences of these specificity alterations remained unknown (Rodríguez et al. 2014).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Rodríguez et al. 2014 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    In astrocytes, CD36 has been implicated as a novel mediator of injury-induced astrogliosis and scar formation in stroke, with the hypothesis that CD36 is required for GFAP expression.

    “In astrocytes, the inflammatory mediator CD36 has been implicated as a novel mediator of injury‐induced astrogliosis and scar formation in stroke, with the hypothesis that CD36 is required for GFAP expression (Bao et al. 2012).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Bao et al. 2012 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    AQP4 is abundant in the endfeet of astrocytes that are in contact with cerebral vasculature and pia; this isoform of aquaporin, an integral transmembrane protein, is involved with water-conduction and our levels of expression are associated with reactive gliosis.

    “AQP4 is abundant in the endfeet of astrocytes that are in contact with cerebral vasculature and pia; this isoform of aquaporin, an integral transmembrane protein, is involved with water‐conduction and our levels of expression are associated with reactive gliosis (Owasil et al. 2020; Dai et al. 2023).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Owasil et al. 2020 source
    • cites Dai et al. 2023 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    A mouse model of brain oxysterol imbalance, especially elevated levels of 27-hydroxycholesterol, displays a downregulation of glutamate transporters in the hippocampus.

    “In this regard, recently, it was reported that a mouse model of brain oxysterol imbalance, especially the elevated levels of 27‐hydroxycholesterol, displays a downregulation of glutamate transporters in the hippocampus (Spanos et al. 2024).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Spanos et al. 2024 source

  • Probably not a substantial claim. It may state two separate claims as one.

    Premisecitation

    Marschallinger et al. (2020) demonstrated that a novel, age-associated, LD-rich microglial cell, "LDAM," exhibits an accumulation of dysfunctional lysosomal deposits, impaired phagocytosis, and release of pro-inflammatory mediators.

    “We performed this labeling because Marschallinger et al. (2020) demonstrated that a novel, age‐associated, LD‐rich microglial cell, “LDAM,” exhibits an accumulation of dysfunctional lysosomal deposits, impaired phagocytosis, and release of pro‐inflammatory mediators.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Marschallinger et al. (2020) source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Study design and conductnone given

    The study strictly limited the use of the cells to a maximum of five passages after thawing and confirmed that the line is not listed in the ICLAC Register of Misidentified Cell Lines.

    “To mitigate the risk of phenotypic drift, we strictly limited the use of the cells to a maximum of five passages after thawing and confirmed that the line is not listed in the ICLAC Register of Misidentified Cell Lines.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.

    Premisecitation

    S100B is expressed mainly in astrocytes and is an essential cytoskeleton component.

    “Expressed mainly in astrocytes, S100B is an essential cytoskeleton component (Barateiro et al. 2016).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Barateiro et al. 2016

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Compared with neurons, astrocytes are relatively resistant to oxidative stress.

    “Compared with neurons, astrocytes are relatively resistant to oxidative stress (Bélanger et al. 2011; Wilson 1997).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Bélanger et al. 2011 source
    • cites Wilson 1997 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    In vitro studies previously published by our group using endothelial cells and macrophages demonstrated increased reactive species generation after 1 h of exposure to oxidized LDL.

    “In vitro studies previously published by our group using endothelial cells (Hort et al. 2014) and macrophages (Straliotto et al. 2013) also demonstrated increased reactive species generation after 1 h of exposure to oxidized LDL.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Hort et al. 2014 source
    • cites Straliotto et al. 2013 source

  • It may state two separate claims as one.

    Interpretationreasoning

    LDL exposure impacts astrocytic reactivity and neurotransmitter homeostasis.

    “These metabolic alterations are accompanied by increased GFAP gene expression and changes in glutamate transporter expression, suggesting that LDL exposure impacts astrocytic reactivity and neurotransmitter homeostasis.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Premisecitation

    Prior studies have shown that LDLr−/− mice already exhibit BBB dysfunction, neuroinflammation, astrogliosis, oxidative stress, and memory deficits at the young adult stage.

    “Notably, our prior studies have shown that LDLr−/− mice already exhibit BBB dysfunction, neuroinflammation, astrogliosis, oxidative stress, and memory deficits at this young adult stage (de Oliveira et al. 2014; de Oliveira, Engel, de Paula, Dos Santos, et al. 2020; de Oliveira, Engel, de Paula, Melo, et al. 2020).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites de Oliveira et al. 2014 source
    • cites de Oliveira, Engel, de Paula, Dos Santos, et al. 2020

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites de Oliveira, Engel, de Paula, Melo, et al. 2020

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim. It may state two separate claims as one.

    Premisecitation

    The scavenger receptor family recognizes danger-associated molecular patterns and expression of CD36 mediates cell-specific responses such as inflammatory responses, phagocytosis of apoptotic cells, fatty acid transport across the BBB, and lipid uptake, leading to activation of PPARγ transcriptional pathways.

    “The scavenger receptor family recognizes danger‐associated molecular patterns and expression of CD36 mediates cell‐specific responses such as inflammatory responses, phagocytosis of apoptotic cells, fatty acid transport across the BBB, and lipid uptake, leading to activation of PPARγ transcriptional pathways (Canton et al. 2013; Silverstein and Febbraio 2009; Ioghen et al. 2021).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites Canton et al. 2013 source
    • cites Silverstein and Febbraio 2009 source
    • cites Ioghen et al. 2021 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    A previous study demonstrated that 27-OHC at concentrations starting at 10 μM induced early and late apoptosis in C6 cells.

    “A previous study evaluated cell death parameters in C6 cells after incubation with 27‐OHC and demonstrated that starting at a concentration of 10 μM, 27‐OHC induced early and late apoptosis in C6 cells (An et al. 2017).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites An et al. 2017

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim. It may be too vague to check against evidence. The paper’s conclusions may not depend on it.

    Study design and conductnone given

    The direct functional consequences of the morphological changes for neuronal health and cognition remain to be established and require further investigation.

    “Furthermore, while these morphological changes are significant, their direct functional consequences for neuronal health and cognition remain to be established and require further investigation.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Previous studies have identified age-dependent heterogeneity in astrocytic reactivity profiles.

    “Previous studies have reported increased astrocyte density in the hippocampus of LDLr−/− mice (de Oliveira et al. 2014; de Oliveira, Engel, de Paula, Dos Santos, et al. 2020) and have identified age‐dependent heterogeneity in astrocytic reactivity profiles (Clarke et al. 2018; Lee et al. 2022).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Clarke et al. 2018 source
    • cites Lee et al. 2022 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    CD36 has been studied in inflammatory conditions such as atherosclerosis, ischemic brain injury, and AD.

    “CD36, a class B scavenger receptor also known as fatty acid translocase, has been studied in inflammatory conditions such as atherosclerosis, ischemic brain injury, and AD (Kim et al. 2008).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Kim et al. 2008 source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Interpretationreasoning

    The acute exposure paradigm used in the experiments may contribute to the lack of statistical significance observed in some effects.

    “Beyond the model choice, the acute exposure paradigm used in our experiments may also contribute to the lack of statistical significance observed in some effects.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: Astrocytic responses to chronic lipid challenges may require longer periods to fully manifest.

  • Probably not a substantial claim. It may state two separate claims as one.

    Study design and conductreasoning

    By combining these two time points, the present study allows assessment of the temporal evolution of astrocytic responses to hypercholesterolemia, distinguishing early alterations from age-dependent changes that may contribute to cognitive decline in middle-aged FH.

    “By combining these two time points, the present study allows assessment of the temporal evolution of astrocytic responses to hypercholesterolemia, distinguishing early alterations from age‐dependent changes that may contribute to cognitive decline in middle‐aged FH.”
    Quote in the paperSubstantial claim
76 of 76

  • The paper relies on this without support.

    LDL cholesterol reaches astrocytes in the brain in the in vivo hypercholesterolemia models used.

    “Therefore, BBB dysfunction may permit the entry of LDL cholesterol into the CNS under peripheral inflammatory conditions, such as hypercholesterolemia and metabolic disease.”

  • The paper relies on this without support.

    The C6 rat astroglial cell line is a valid model for studying astrocyte biology.

    “The high‐passage C6 rat cell line (Rattus norvegicus; RRID:CVCL_0194) was cultured in DMEM low glucose media containing 26.2 mM NaHCO3, 40 μ/mL penicillin‐streptomycin, 40 μg/mL gentamicin, 0.25 μg/mL fungizone (amphotericin B), 8.39 mM HEPES buffer, sodium salt, and supplemented with 5% FBS at 37°C and 5% CO2 using standard cell culture methods.”

  • The paper relies on this without support.

    The LDL cholesterol concentrations used in vitro (50 and 300 μg/mL) are physiologically relevant to the brain.

    “then the cells were treated with two different concentrations of LDL cholesterol (50 and 300 μg/mL) or phosphate‐buffered saline (PBS) ‐ control (Ctrl) for 24 or 48 h.”

  • The paper relies on this without support.

    The isolated human LDL cholesterol used in vitro is not oxidized and is biologically representative of native LDL.

    “To 35 mL of human plasma, 0.5% saccharose was added to prevent LDL aggregation, 1 mg/mL EDTA to prevent LDL oxidation, and 0.326 g/mL KBr to adjust the density.”

  • The paper relies on this without support.

    The LDLr−/− mouse model used here develops brain exposure to LDL cholesterol relevant to hypercholesterolemia.

    “In vivo, hippocampal astrocytes from young (3‐month‐old) and middle‐aged (14‐month‐old) LDL receptor knockout (LDLr−/−) and wild‐type C57BL/6 mice were analyzed by immunofluorescence and quantitative reverse transcription polymerase chain reaction (RT‐qPCR).”

  • The paper relies on this without support.

    Changes in mRNA levels of astrocytic genes reflect corresponding changes in protein levels or functional astrocyte states.

    “Aging and genotype influenced astrocyte morphology and expression of genes such as S100 calcium‐binding protein B (S100B) and aquaporin‐4 (AQP4).”

  • The paper relies on this without support.

    Hypercholesterolemia-induced impairment of the BBB may trigger extravasation of LDL cholesterol into the brain parenchyma, which could harm cell homeostasis.

    “In this sense, we believe that the impairment of the BBB that can occur due to hypercholesterolemia may trigger the extravasation of LDL cholesterol into the brain parenchyma, which could harm cell homeostasis.”

  • The paper relies on this without support.

    LDL particles carry other lipids, including triglycerides and phospholipids, in addition to cholesterol and cholesteryl esters.

    “In addition to cholesterol and cholesteryl esters, LDL particles also carry other lipids, including triglycerides and phospholipids.”

  • The authors state this as an assumption.

    Physiological LDL concentrations in the brains of humans or mice with hypercholesterolemia are unknown, because LDL is not normally found in the CNS and its concentration under pathological BBB disruption has not been quantified.

    “However, such values remain unknown, as LDL is not normally found in the CNS and its concentration under pathological conditions of BBB disruption has not yet been quantified.”

  • The paper relies on this without support.

    Astrocytic responses to chronic lipid challenges may require longer periods to fully manifest.

    “Given that astrocytic responses to chronic lipid challenges may require longer periods to fully manifest, future studies will include extended incubation times to better characterize the long‐term effects of LDL on redox balance and metabolic function.”
10 of 10

Statistics recomputed

Every reported test result, recomputed from its test statistic and degrees of freedom. A result shows as not adding up only once an independent check has confirmed it.

  • Not confirmedF(1, 11) = 3.935reported p = .072 · recomputed p = .073

    “F(1,11) = 3.935, p = .072”

  • ConsistentF(2, 6) = 99reported p < .0001 · recomputed p < .001

    “F(2, 6) = 99.00, p < .0001”

  • ConsistentF(2, 6) = 6.334reported p = .0332 · recomputed p = .033

    “F(2, 6) = 6.334, p = .0332”

  • ConsistentF(2, 6) = 0.1155reported p = .8929 · recomputed p = .893

    “F(2, 6) = 0.1155, p = .8929”

  • ConsistentF(2, 6) = 0.0192reported p = .9811 · recomputed p = .981

    “F(2, 6) = 0.01915, p = .9811”

  • ConsistentF(2, 9) = 13.07reported p = .0022 · recomputed p = .002

    “F(2, 9) = 13.07, p = .0022”

  • ConsistentF(2, 9) = 16.6reported p = .0009 · recomputed p < .001

    “F(2, 9) = 16.6, p = .0009”

  • ConsistentF(2, 8) = 21.25reported p = .0006 · recomputed p < .001

    “F(2, 8) = 21.25, p = .0006”

  • ConsistentF(2, 9) = 2.987reported p = .1012 · recomputed p = .101

    “F(2, 9) = 2.987, p = .1012”

  • ConsistentF(2, 8) = 1.45reported p = .2903 · recomputed p = .290

    “F(2, 8) = 1.450, p = .2903”

  • ConsistentF(2, 9) = 2.415reported p = .1446 · recomputed p = .145

    “F(2, 9) = 2.415, p = .1446”

  • ConsistentF(2, 8) = 2.596reported p = .1353 · recomputed p = .135

    “F(2, 8) = 2.596, p = .1353”

  • ConsistentF(2, 8) = 0.4551reported p = .6499 · recomputed p = .650

    “F(2, 8) = 0.4551, p = .6499”

  • ConsistentF(2, 9) = 60.7reported p < .0001 · recomputed p < .001

    “F(2, 9) = 60.70, p < .0001”

  • ConsistentF(2, 9) = 4.874reported p = .0413 · recomputed p = .037

    “F(2, 9) = 4.874, p = .0413”

    The paper states a correction for multiple tests

  • ConsistentF(2, 6) = 142reported p < .0001 · recomputed p < .001

    “F(2, 6) = 142.0, p < .0001”

  • ConsistentF(2, 6) = 0.2799reported p = .7652 · recomputed p = .765

    “F(2, 6) = 0.2799, p = .7652”

  • ConsistentF(2, 6) = 2.602reported p = .1535 · recomputed p = .154

    “F(2, 6) = 2.602, p = .1535”

  • ConsistentF(2, 6) = 0.7868reported p = .4972 · recomputed p = .497

    “F(2, 6) = 0.7868, p = .4972”

  • ConsistentF(2, 6) = 6.956reported p = .0274 · recomputed p = .027

    “F(2, 6) = 6.956, p = .0274”

  • ConsistentF(2, 6) = 9.537reported p = .0137 · recomputed p = .014

    “F(2, 6) = 9.537, p = .0137”

  • ConsistentF(2, 9) = 3.253reported p = .0865 · recomputed p = .086

    “F(2, 9) = 3.253, p = .0865”

  • ConsistentF(2, 8) = 15.18reported p = .0019 · recomputed p = .002

    “F(2, 8) = 15.18, p = .0019”

  • ConsistentF(2, 6) = 3.628reported p = .0927 · recomputed p = .093

    “F(2, 6) = 3.628, p = .0927”

  • ConsistentF(2, 6) = 0.1157reported p = .8927 · recomputed p = .893

    “F(2, 6) = 0.1157, p = .8927”

  • ConsistentF(2, 6) = 1.887reported p = .2313 · recomputed p = .231

    “F(2, 6) = 1.887, p = .2313”

  • ConsistentF(2, 6) = 1.197reported p = .3653 · recomputed p = .365

    “F(2, 6) = 1.197, p = .3653”

  • ConsistentF(2, 6) = 1.353reported p = .3273 · recomputed p = .327

    “F(2, 6) = 1.353, p = .3273”

  • ConsistentF(1, 11) = 10.704reported p < .05 · recomputed p = .007

    “F(1,11) = 10.704, p < .05”

  • ConsistentF(1, 16) = 3.9529reported p = .06418 · recomputed p = .064

    “F(1,16) = 3.9529, p = .06418”

  • ConsistentF(1, 16) = 4.3338reported p = .05378 · recomputed p = .054

    “F(1,16) = 4.3338, p = .05378”

  • ConsistentF(1, 16) = 6.1514reported p = .02464 · recomputed p = .025

    “F(1,16) = 6.1514, p = .02464”

  • ConsistentF(1, 16) = 34.043reported p = .00003 · recomputed p < .001

    “F(1,16) = 34.043, p = .00003”

33 of 33

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Statistical tendencies are marked above the stated cutoff

    Minor

    The paper defines a statistical tendency as p ≤ 0.1, yet marks p = 0.1030 and p = 0.1123 with the tendency symbol (#).

  • Sample-size calculation is both described and denied

    Minor

    The methods describe a power-based calculation that indicated a target of approximately six animals per condition, but later state that no formal sample-size calculations were conducted for any animal procedure.

2 of 2

Citations

Cites

Flagged first, then by citations

Research works printed but not linked: 2

  1. 10.3791/2597 · DOI
  2. Sholl D. A., 1953, Dendritic Organization in the Neurons of the Visual and Motor Cortices of the Cat, Journal of Anatomy, 87, 387

Cited by

Sorted by citations

Related

Metadata

Authors
Baumart, G. J., Rodrigues, M. S., Sholl, J. N., de Sousa, A. C., Weber, A. F., Costa‐Beber, L. C., Farias, H. R., Costa, M. V., Peres, A. M., de Camargo, P. R., Fróes, F. T., Bast, R. K., de Bem, A. F., Figueiró, F., Guma, F. T. C. R., & de Oliveira, J.
Affiliations
  • Universidade Federal do Rio Grande do Sul (BR)
  • Graduate Program in Biological Sciences: Biochemistry, Department of Biochemistry, Institute of Health Basic Sciences Universidade Federal do Rio Grande do Sul (UFRGS) Porto Alegre Rio Grande do Sul Brazil
  • Graduate Program in Molecular Pathology, School of Medicine University of Brasília (UnB) Brasília Brazil
Corresponding author
de Oliveira, Jade
ORCID
Venue
Journal of Neurochemistry
Publisher
Wiley
Published
2026-03
Volume
170
Issue
3
Pages
e70403-e70403
ISSN
  • 0022-3042 (ISSN-L)
  • 1471-4159
DOI
10.1111/jnc.70403
PMC
PMC12982915
OpenAlex
W7135238771
Cited by
2
References
98
Citations by year
2026: 2
FWCI
4.35
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Neurochemistry (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • Conselho Nacional de Desenvolvimento Científico e Tecnológico (407006/2021‐4)
  • Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (21/2551‐0000740‐0)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Cholesterol and Lipid Metabolism
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
Keywords
LDL cholesterol, astrocytes, hypercholesterolemia, lipid accumulation, LDL receptor, HMGCR, SREBF1, glial fibrillary acidic protein, fatty acid uptake, CD36, superoxide dismutase, oxidative stress, Nrf2, astrocyte morphology, astrocyte proliferation, astrocyte metabolism, hippocampus, astrocyte-neuron interaction, neurodegenerative diseases
MeSH terms
Animals, Astrocytes, Hippocampus, Humans, Cholesterol, LDL, Male, Mice, Inbred C57BL, Lipid Metabolism, Mice, Rats

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

A language model drew the claims from the paper on 2026-11-10, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).

A second model labelled each claim on 2026-11-10 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).

Cited sources are checked with a language model’s help (gpt-6-sol).

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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