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Cited by 32RNA Research and SplicingBerberine and alkaloids researchCholesterol and Lipid Metabolism

The DOI matches its Crossref record; 40 of the 40 references it cites were found in a registry or an index; none of the 44 sources its claims cite is checked for support yet.

The Critical Role of mRNA Destabilizing Protein Heterogeneous Nuclear Ribonucleoprotein D in 3′ Untranslated Region–Mediated Decay of Low-Density Lipoprotein Receptor mRNA in Liver Tissue

Open access
Singh, Amar Bahadur, Li, Hai, Kan, Chin Fung Kelvin, Dong, Bin, Nicolls, Mark R., Liu, Jingwen·2014·Arteriosclerosis, Thrombosis, and Vascular Biology
DOI 10.1161/atvbaha.112.301131PMC PMC4032120
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Abstract

Objective—: Previous studies showed that low-density lipoprotein receptor (LDLR) mRNA 3′ untranslated region (UTR) contains regulatory elements responsible for rapid mRNA turnover in hepatic cells and mediates the mRNA stabilization induced by berberine (BBR). Here, we elucidate the underlying mechanism of BBR’s action by characterizing mRNA-binding proteins that modulate LDLR mRNA decay via 3′UTR in liver tissue in vivo. Approach and Results—: We generated a transgenic mouse model (Alb-Luc-UTR) that expresses Luc-LDLR3′UTR reporter gene driven by the albumin promoter to study 3′UTR function in mediating LDLR mRNA decay in liver tissue. We show that treating Alb-Luc-UTR mice with BBR led to significant increases in hepatic bioluminescence signals, Luc-UTR mRNA, and LDLR mRNA levels as compared with control mice. These effects were accompanied by specific reductions of mRNA decay-promoting factor heterogeneous nuclear ribonucleoprotein D (hnRNP D) in liver of BBR-treated mice. Knockdown and overexpression studies further demonstrated that hnRNP D p37 isoform plays a major role in promoting hepatic LDLR mRNA degradation. In addition, we examined LDLR mRNA half-life, Luc-UTR reporter activity, and hnRNP D expression levels in cell lines derived from extrahepatic tissues. We demonstrated that strengths of 3′UTR in promoting mRNA degradation correlate with hnRNP D cellular abundances in nonhepatic cell lines, thereby suggesting its involvement in LDLR mRNA degradation beyond liver tissue. Conclusions—: hnRNP D is critically involved in LDLR mRNA degradation in liver tissue in vivo. The inverse relationship of hnRNP D abundance with LDLR mRNA levels after BBR treatment suggests the potential of hnRNP D of being a novel therapeutic target for LDL cholesterol lowering.

Overview

AI snapshot

Written from the paper’s full text.

TL;DR

This study generated an albumin-promoter-driven Luc-LDLR 3′UTR reporter mouse to show that berberine increases hepatic LDLR mRNA through the 3′UTR and reduces hnRNP D. hnRNP D p37 is identified as a key destabilizing factor for LDLR mRNA in liver tissue, with potential as an LDL-lowering target.

Key findings

  • In Alb-Luc-UTR mice, berberine (200 mg/kg/day for 14 days) increased hepatic bioluminescence ≈2-fold (P<0.05), Luc-UTR mRNA 2-fold (P<0.05), and endogenous mouse LDLR mRNA 30% (P<0.01), without changing albumin mRNA or SREBP pathway genes.
  • Berberine reduced total hnRNP D mRNA by 20% (P<0.05) and the prominent p37 isoform protein by ≈37% (P<0.05), while hnRNP I and KSRP were unchanged; acute 3-day treatment increased LDLR mRNA 45% and decreased hnRNP D mRNA/protein ≈25%.
  • hnRNP D knockdown in mouse primary hepatocytes reduced hnRNP D mRNA by 75% and p37 protein by 60% at 50 MOI, and increased LDLR mRNA by 50% and 75% at 20 and 50 MOI, respectively.
  • LDLR mRNA half-lives were longer in nonhepatic A7r5 (8.4±0.5 h) and H9c2 (4.1±0.1 h) cells than in HepG2 (1.5±0.1 h) and Hepa 1–6 (2.7±0.2 h) cells, correlating with lower hnRNP D levels.

Methods

  • Transgenic FVB mice expressing an albumin promoter-driven Luc-LDLR 3′UTR reporter were treated with berberine (200 mg/kg/day by oral gavage) for 14 days (n=10 per group) or 3 days; liver was analyzed by in vivo bioluminescence, qRT-PCR, and Western blotting.
  • Cell studies used HepG2, Hepa 1–6, mouse primary hepatocytes, H9c2, A7r5, and Y1 cells with LDLR3′UTR reporter assays, ARE constructs, adenoviral shRNA knockdown of hnRNP D, isoform overexpression, actinomycin D mRNA half-life measurements, and miRNA arrays/anti-miRNA validation.
  • Materials and Methods are available in the online-only Supplement.

Limitations

  • Stated: the reason for the weaker berberine effect on endogenous LDLR mRNA versus Luc-UTR transgene was unclear; further studies are required to understand hnRNP D’s role in berberine-mediated LDLR mRNA stabilization in HepG2 cells, including possible phosphorylation/binding changes; defining a BBR-independent mechanism regulating hnRNP D expression is a future question.
  • Evident: the main text states that Materials and Methods and several key data are in the online-only Supplement, limiting what can be assessed from the paper text alone.
  • Evident: in vivo evidence for hnRNP D is largely correlative (berberine lowers hnRNP D while LDLR mRNA rises), whereas direct hnRNP D knockdown/overexpression was performed ex vivo or in cell lines, not in mice.

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

Integrity signals

Registry

Crossref · 10.1161/atvbaha.112.301131

Cites

40 of 40 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Verification

Claims the paper makes

0 of 44 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Decay rates of cyclin D1 mRNA did not show substantial differences in these cell lines and mouse primary hepatocytes.

    “In the mRNA decay experiments conducted in these cell lines and mouse primary hepatocytes, we measured the decay rates of cyclin D1 mRNA, which did not show substantial differences (Figure V in the online-only Data Supplement).”
    Quote in the paperNot a substantial claim
  • Findingown data

    Alignment of cDNA sequences of mouse and human hnRNP D isoforms identified a 29-bp region that is identical in all sequences.

    “Alignment of cDNA sequences of mouse and human hnRNP D isoforms identified a 29-bp region that is identical in all sequences (Figure IIB in the online-only Data Supplement).”
    Quote in the paperSubstantial claim
  • Findingown data

    Inclusion of full-length LDLR 3′UTR led to an 80% reduction in luciferase activity in Hepa 1-6 cells.

    “Inclusion of full-length UTR led to an 80% reduction in luciferase activity.”
    Quote in the paperSubstantial claim
  • Findingown data

    Acute BBR treatment for 3 days increased Luc-UTR transgene expression in Alb-Luc-UTR mice.

    “Examination of bioluminescence signal (Figure 4A) and the luciferase activity of isolated liver tissues (Figure 4B) confirmed the increased expression of Luc-UTR transgene in BBR-treated mice”
    Quote in the paperSubstantial claim
  • Findingown data

    Liver LDLR protein levels in BBR-treated mice were slightly increased with statistical significance.

    “The liver LDLR protein levels in BBR-treated mice were slightly increased with statistic significance.”
    Quote in the paperSubstantial claim
  • It may state two separate claims as one. 20
  • Probably not a substantial claim. 12

  • It may state two separate claims as one.

    Findingown data

    hnRNP D mRNA and protein levels were specifically reduced by BBR treatment in Alb-Luc-UTR mice, which correlated with elevated endogenous LDLR mRNA and Luc-UTR mRNA levels.

    “We have now demonstrated that the mRNA and protein levels of hnRNP D were specifically reduced by BBR treatment in Alb-Luc-UTR mice, which correlated with elevated endogenous LDLR mRNA and Luc-UTR mRNA levels.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    In Hepa 1-6 cells, hnRNP D p42 and p45 isoforms displayed modest activities, whereas p42 did not repress reporter activity in HepG2 cells.

    “In Hepa 1–6 cells, hnRNP D p42 and p45 isoforms displayed modest activities, whereas p42 did not repress the reporter activity in HepG2 cells.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Acute BBR treatment led to approximately 25% decrease in hnRNP D mRNA and protein levels without affecting KSRP and hnRNP I mRNA.

    “the transient administration of BBR led to ≈25% decrease in hnRNP D mRNA (Figure 4D) and protein levels (Figure 4E) without affecting mRNA expressions of KSRP and hnRNP I.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    The half-lives of LDLR transcript in A7r5 and H9c2 cells were 8.4±0.5 and 4.1±0.1 hours, considerably longer than in HepG2 cells (1.5±0.1 hours) and Hepa 1-6 cells (2.7±0.2 hours).

    “The results showed that the half-lives of LDLR transcript in A7r5 and H9c2 cells were 8.4±0.5 and 4.1±0.1 hours, which were considerably longer than the half-life of LDLR mRNA in HepG2 cells (1.5±0.1 hours) and Hepa 1–6 cells (2.7±0.2 hours), respectively.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Ad-shHNRNPD infection did not affect mRNA levels of hnRNP I, KSRP, or SREBP2 target genes HMGCR and PCSK9 in mouse primary hepatocytes.

    “mRNA levels of other mRNA-binding proteins (hnRNP I, KSRP) as well as SREBP2 target genes (HMGCR, PCSK9) in mouse primary hepatocytes were not affected by Ad-shHNRNPD infection (Figure 5C)”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    At 50 multiplicity of infection, hnRNP D mRNA level was reduced by 75%, p37 protein was reduced by 60%, p45 was decreased below detection, and LDLR protein levels were increased in Ad-shHNRNPD-infected hepatocytes.

    “At 50 multiplicity of infection, hnRNP D mRNA level was reduced by 75% (Figure 5A), the protein amount of p37 isoform was reduced by 60% as compared with cells infected with control virus Ad-GFP, and the p45 isoform was decreased to a level of below detection, which was accompanied by increased LDLR protein levels in Ad-shHNRNPD–infected hepatocytes (Figure 5B).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    In HepG2 cells, hnRNP D protein levels were markedly reduced in Ad-shHNRNPD-infected cells, accompanied by a 37% increase in LDLR protein abundance.

    “hnRNP D protein levels were markedly reduced in Ad-shHNRNPD–infected HepG2 cells, which was accompanied with 37% increase in LDLR protein abundance.”
    Quote in the paperSubstantial claim

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

    Findingown data

    miR-1244 and miR-1979 have no binding sites in LDLR3′UTR but were substantially downregulated by BBR.

    “Two miRNAs, miR-1244 and miR-1979, have no binding sites in LDLR3′UTR but they were substantially downregulated by BBR, thus they were included in our validation studies.”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    Exogenous expression of p37 or p40 in H9c2 cells did not affect pLuc control reporter activity but reduced pLuc-LDLR3′UTR luciferase activity to 57% and 45% of control.

    “Expressions of either isoform did not affect the pLuc control reporter activity but reduced pLuc-LDLR3′UTR Luc activity to 57% and 45% of control.”
    • Also stated in: § Discussion · boosted
      “By cotransfection of p37 or p40 isoform with pLuc-LDLR3′UTR construct into H9c2 cells, we further demonstrated that exogenous expression of each isoform accelerated luciferase mRNA degradation in cardiomyocytes.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Exogenous expression of p37 or p40 markedly reduced luciferase activity by approximately 80% in Hepa 1-6 cells and by 50% to 60% in HepG2 cells.

    “exogenous expression of p37 or p40 markedly reduced luciferase activity by ≈80% in Hepa 1–6 cells and by 50% to 60% in HepG2 cells.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    BBR administration in Alb-Luc-UTR mice either chronically (14 days) or acutely (3 days) elevated bioluminescent signals and increased mRNA levels of Luc-UTR transcript and endogenous LDLR mRNA in liver.

    “BBR administration in Alb-Luc-UTR mice either chronically (14 days; Figure 2) or acutely (3 days; Figure 4) elevated bioluminescent signals and increased mRNA levels of Luc-UTR transcript and endogenous LDLR mRNA in liver.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Mouse liver tissues and primary hepatocytes express the predominant p37 isoform and the minor p45 form; Hepa 1-6 cells have a low p42/40 band.

    “Mouse liver tissues and primary hepatocytes express the predominant p37 isoform and the minor p45 form. A low signal of p42/40 band was detected in Hepa 1–6 cells.”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    Overexpression of p37 and p42 reduced luciferase activity in control cells and lowered the induction of luciferase activity by BBR treatment.

    “The results showed that p37 and p42 overexpression not only reduced the luciferase activity in control cells but they also lowered the induction of luciferase activity by BBR treatment (Figure IIIC in the online-only Data Supplement)”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Increases of Luc-UTR transgene and LDLR mRNA levels occurred in the absence of changes in albumin mRNA levels or activation of SREBP pathways.

    “Importantly, increases of Luc-UTR transgene and LDLR mRNA levels occurred in the absence of changes in albumin mRNA levels or the activation of SREBP pathways.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Inclusion of all 3 AREs to the coding region of β-globin mRNA resulted in further destabilization of the β-globin fusion transcripts.

    “whereas inclusion of all 3 AREs to the coding region of β-globin mRNA resulted in further destabilization of the β-globin fusion transcripts.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [20] Knouff C Malloy S Wilder J Altenburg MK Maeda N Doubling expression of the low density lipoprotein receptor by truncation of the 3′-untranslated region sequence ameliorates type iii hyperlipoproteinemia in mice expressing the human apoe2 isoform J Biol Chem 2001 276 3856 3862 11076954

  • Probably not a substantial claim.

    Premisecitation

    Previous work identified hnRNP D, hnRNP I, and KSRP as LDLR ARE-BPs.

    “Our previous work conducted in HepG2 cells have identified hnRNP D, hnRNP I, and KSRP as LDLR ARE-BPs.25”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [25] Li H Chen W Zhou Y Abidi P Sharpe O Robinson WH Liu J Identification of mRNA-binding proteins that regulate the stability of LDL receptor mRNA through AU-rich elements J Lipid Res 2009 50 820 831 19141871 PMC2666168

  • It may state two separate claims as one.

    Premisecitation

    Berberine increased LDLR mRNA half-life nearly 3-fold without affecting gene transcription in HepG2 cells.

    “increased LDLR mRNA half-life nearly 3-fold without affecting gene transcription in HepG2.15”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [15] Kong W Wei J Abidi P Lin M Inaba S Li C Wang Y Wang Z Si S Pan H Wang S Wu J Li Z Liu J Jiang J-D Berberine is a promising novel cholesterol-lowering drug working through a unique mechanism distinct from statins Nat Med 2004 10 1344 1352 15531889

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

    Premisecitation

    Some ARE-BPs are decay-promoting factors, such as KSRPs, that interact with AREs and recruit RNA degradation machinery to the mRNA.

    “Some ARE-BPs are decay-promoting factors, such as KH-type splicing regulatory proteins (KSRPs), that interact with AREs and recruit RNA degradation machinery to the mRNA.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [19] Briata P Chen CY Giovarelli M Pasero M Trabucchi M Ramos A Gherzi R KSRP, many functions for a single protein Front Biosci (Landmark Ed) 2011 16 1787 1796 21196264

  • Probably not a substantial claim.

    Premisecitation

    The liver removes more than 70% to 80% of LDL-c from circulation via LDLR-mediated endocytosis.

    “The liver is the organ responsible for removing >70% to 80% of LDL-c from circulation via LDL receptor (LDLR)–mediated endocytosis.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [3] Spady DK Hepatic clearance of plasma low density lipoproteins Semin Liver Dis 1992 12 373 385 1465622
    • cites [4] Brown MS Goldstein JL A receptor-mediated pathway for cholesterol homeostasis Science 1986 232 34 47 3513311
    • cites [5] Goldstein JL Brown MS Regulation of the mevalonate pathway Nature 1990 343 425 430 1967820

  • It may state two separate claims as one.

    Interpretationown data

    3′UTR regulates LDLR mRNA stability with cell-type/tissue specificity, and variable expression levels of hnRNP D (particularly p37) contribute to the strength of 3′UTR-mediated LDLR mRNA degradation in a particular cell type or tissue.

    “Collectively, these data obtained from various cell lines suggest that 3′UTR regulates LDLR mRNA stability with cell-type specificity and variable expression levels of hnRNP D in different cell types might be an underlying factor contributing to the strength of 3′UTR in LDLR mRNA degradation in a particular cell type or a tissue.”
    • Also stated in: § Discussion · hedged
      “These in vitro studies suggest that 3′UTR regulates LDLR mRNA stability with cell-type/tissue specificity and its activity positively correlates with cellular levels of hnRNP D p37 isoform.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Interpretationown data

    Rises in LDLR mRNA level occurred concomitantly with declines of cellular abundance of hnRNP D protein, providing the first in vivo evidence linking this decay-promoting ARE-BP to LDLR mRNA destabilization in liver tissue.

    “Collectively, these 2 treatment studies consistently demonstrated that rises in LDLR mRNA level occurred concomitantly with declines of cellular abundance of hnRNP D protein, which provide the first in vivo evidence linking this decay-promoting ARE-BP to LDLR mRNA destabilization in liver tissue.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Premisecitation

    Human LDLR mRNA contains a 2.5-kb-long 3′UTR in which 3 AU-rich elements were identified and shown to mediate rapid turnover of LDLR mRNA in liver cells.

    “Human LDLR mRNA contains a 2.5-kb-long stretch of 3′UTR16 in which 3 AU-rich elements (AREs) were previously identified and shown to mediate the rapid turnover rate of LDLR mRNA in liver cells.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [12] Wilson GM Vasa MZ Deeley RG Stabilization and cytoskeletal- association of LDL receptor mRNA are mediated by distinct domains in its 3′ untranslated region J Lipid Res 1998 39 1025 1032 9610769
    • cites [13] Wilson GM Roberts EA Deeley RG Modulation of LDL receptor mRNA stability by phorbol esters in human liver cell culture models J Lipid Res 1997 38 437 446 9101425
    • cites [16] Yamamoto T Davis CG Brown MS Schneider WJ Casey ML Goldstein JL Russell DW The human LDL receptor: a cysteine-rich protein with multiple Alu sequences in its mRNA Cell 1984 39 27 38 6091915

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

    Premisecitation

    The LDL-c lowering effect of BBR has been demonstrated in multiple clinical studies.

    “The LDL-c lowering effect of the herbal medicine BBR has been demonstrated in multiple clinical studies across the globe.22–24,37,38”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [22] Zhang Y Li X Zou D Liu W Yang J Zhu N Huo L Wang M Hong J Wu P Ren G Ning G Treatment of type 2 diabetes and dyslipidemia with the natural plant alkaloid berberine J Clin Endocrinol Metab 2008 93 2559 2565 18397984
    • cites [23] Yin J Xing H Ye J Efficacy of berberine in patients with type 2 diabetes mellitus Metabolism 2008 57 712 717 18442638 PMC2410097
    • cites [24] Wei W Zhao H Wang A Sui M Liang K Deng H Ma Y Zhang Y Zhang H Guan Y A clinical study on the short-term effect of berberine in comparison to metformin on the metabolic characteristics of women with polycystic ovary syndrome Eur J Endocrinol 2012 166 99 105 22019891
    • cites [37] Derosa G D’Angelo A Bonaventura A Bianchi L Romano D Maffioli P Effects of berberine on lipid profile in subjects with low cardiovascular risk Expert Opin Biol Ther 2013 13 475 482 23441841
    • cites [38] Pisciotta L Bellocchio A Bertolini S Nutraceutical pill containing berberine versus ezetimibe on plasma lipid pattern in hypercholesterolemic subjects and its additive effect in patients with familial hypercholesterolemia on stable cholesterol-lowering treatment Lipids Health Dis 2012 11 123 22998978 PMC3477057

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

    Premisecitation

    AREs are the best characterized sequence determinants of messenger stability among known RNA cis-regulatory elements.

    “AREs are the best characterized sequence determinants of messenger stability among known RNA cis-regulatory elements.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [17] Zhang T Kruys V Huez G Gueydan C AU-rich element-mediated translational control: complexity and multiple activities of trans-activating factors Biochem Soc Trans 2002 30 pt 6 952 958 12440953

  • It may state two separate claims as one.

    Interpretationown data

    ARE sequences are possibly involved in the mRNA stabilizing process as well as in the rapid degradation process.

    “This suggests that these ARE sequences are possibly involved in the mRNA stabilizing process as well as in the rapid degradation process.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim.

    Premisecitation

    The mouse albumin promoter has been widely used to obtain constitutive target gene expression specifically in liver tissue.

    “The mouse albumin (Alb) promoter has been widely used to obtain constitutive target gene expression specifically in liver tissue.26–28”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [26] Pinkert CA Ornitz DM Brinster RL Palmiter RD An albumin enhancer located 10 kb upstream functions along with its promoter to direct efficient, liver-specific expression in transgenic mice Genes Dev 1987 1 268 276 3678824
    • cites [27] Izban MG Papaconstantinou J Cell-specific expression of mouse albumin promoter. Evidence for cell-specific DNA elements within the proximal promoter region and cis-acting DNA elements upstream of −160 J Biol Chem 1989 264 9171 9179 2722822
    • cites [28] Wu Y Wang YY Nakamoto Y Li YY Baba T Kaneko S Fujii C Mukaida N Accelerated hepatocellular carcinoma development in mice expressing the Pim-3 transgene selectively in the liver Oncogene 2010 29 2228 2237 20101231

  • Probably not a substantial claim.

    Premisecitation

    The destabilizing functions of ARE sequences are achieved through their interaction with ARE-binding proteins.

    “The destabilizing functions of ARE sequences are achieved through their interaction with ARE-binding proteins (ARE-BPs).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [18] Meyer S Temme C Wahle E Messenger RNA turnover in eukaryotes: pathways and enzymes Crit Rev Biochem Mol Biol 2004 39 197 216 15596551

  • Probably not a substantial claim.

    Premisereasoning

    The critical question of which ARE-BP is the key player that modulates LDLR mRNA levels in liver tissue in vivo has not been addressed.

    “the critical question of which ARE-BP is the key player that modulates LDLR mRNA levels in liver tissue in vivo has not been addressed.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    The MAP kinase MEK-ERK signaling pathway is critically involved in BBR's action mechanism.

    “We have previously demonstrated that the MAP kinase MEK-ERK signaling pathway is critically involved in BBR’s action mechanism.15”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [15] Kong W Wei J Abidi P Lin M Inaba S Li C Wang Y Wang Z Si S Pan H Wang S Wu J Li Z Liu J Jiang J-D Berberine is a promising novel cholesterol-lowering drug working through a unique mechanism distinct from statins Nat Med 2004 10 1344 1352 15531889

  • It may state two separate claims as one.

    Interpretationown data

    Knockdown studies in HepG2 cells provided initial evidence that hnRNP D is involved in BBR-induced LDLR mRNA stabilization without significant reductions of hnRNP D protein levels by BBR treatment.

    “Knockdown studies in HepG2 cells provided the initial evidence that hnRNP D is also involved in BBR-induced LDLR mRNA stabilization in this hepatic model cell line without significant reductions of hnRNP D protein levels by BBR treatment (Figure 5E).”
    Quote in the paperSubstantial claim

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

    Premisecitation

    A recent study linked the ERK signaling pathway with nuclear export of the p42 isoform and its stabilizing activity to the 3′UTR of COX-2 mRNA in human synovial fibroblast.

    “A recent study has linked ERK signaling pathway with the nuclear export of p42 isoform and its stabilizing activity to the 3′UTR of COX-2 mRNA in human synovial fibroblast.39”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [39] Zhai B Yang H Mancini A He Q Antoniou J Di Battista JA Leukotriene B(4) BLT receptor signaling regulates the level and stability of cyclooxy-genase-2 (COX-2) mRNA through restricted activation of Ras/Raf/ERK/ p42 AUF1 pathway J Biol Chem 2010 285 23568 23580 20489206 PMC2911275
31 of 31

  • The paper relies on this without support.

    There is a direct correlation of high LDLR expression in liver with low LDL-c level in blood.

    “There is a direct correlation of high expression level of LDLR in liver with low LDL-c level in blood.”

  • The paper relies on this without support.

    Until this study, there was no investigation to validate or challenge the BBR action mechanism under in vivo conditions.

    “However, until this study, there is no investigation to validate or challenge this BBR action mechanism under in vivo conditions.”

  • The paper relies on this without support.

    It was unknown whether hnRNP D, hnRNP I, and KSRP are functionally involved in LDLR mRNA decay in liver tissue.

    “However, it was unknown whether these ARE-BPs are functionally involved in LDLR mRNA decay in liver tissue.”

  • The paper relies on this without support.

    The anti-miRNAs used effectively depleted the targeted miRNAs in HepG2 cells.

    “We used anti-miRNAs to individually deplete these 9 miRNAs in HepG2 cells and examined the effects of knockdown on LDLR3′UTR reporter activity, LDLR mRNA, and protein expressions.”
4 of 4

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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.

  • A7r5 cells are called cardiac cells despite being identified as vascular myocytes

    Minor

    The Results identify A7r5 cells as vascular myocytes, but the Discussion groups them with H9c2 as cardiac cells.

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Metadata

Authors
Singh, A. B., Li, H., Kan, C. F. K., Dong, B., Nicolls, M. R., & Liu, J.
Affiliations
VA Palo Alto Health Care System (US)
ORCID
Venue
Arteriosclerosis, Thrombosis, and Vascular Biology
Publisher
Ovid Technologies (Wolters Kluwer Health)
Published
2014-01
Volume
34
Issue
1
Pages
8-16
ISSN
  • 1079-5642 (ISSN-L)
  • 1524-4636
DOI
10.1161/atvbaha.112.301131
PMC
PMC4032120
OpenAlex
W2048275551
Semantic Scholar
2274cb924bbe12181ce5d90f90702a9eb6fc3302
Cited by
32
References
40
Citations by year
2026: 1 · 2025: 5 · 2024: 4 · 2023: 1 · 2022: 2 · 2021: 5 · 2020: 2 · 2019: 1 · 2018: 2 · 2017: 2 · 2016: 3 · 2014: 4
FWCI
0.77
Type
journal article
Language
en
Open access
Locations
  • Arteriosclerosis Thrombosis and Vascular Biology (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Institutes of Health (5r21at003195-02, 5p01hl108797-10)
  • National Heart, Lung, and Blood Institute (R01HL095686, P01-HL-108797)
  • National Center for Complementary and Integrative Health (R21 AT003195, R01AT006336, 1R21 AT003195-01A2, R01AT002543)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Health Sciences › Medicine › Pharmacology (medical) › Berberine and alkaloids research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Cholesterol and Lipid Metabolism
Keywords
3' untranslated region, mRNA degradation, berberine, liver, LDL cholesterol lowering, mRNA-binding proteins, transgenic mouse model
MeSH terms
Albumins, Animals, Base Sequence, Berberine, Female, Gene Expression Regulation, Half-Life, Humans, Liver, Luciferases, Male, Mice, Inbred C57BL, Mice, Transgenic, Molecular Sequence Data, Promoter Regions, Genetic, Receptors, LDL, RNA, Messenger, Transfection, Genes, Reporter, 3' Untranslated Regions, RNA Stability, Hepatocytes, Heterogeneous-Nuclear Ribonucleoprotein D, RNA Interference, Mice, Rats, Hep G2 Cells

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 2026-11-09, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).

A second model labelled each claim on 2026-11-09 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).

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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