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Cited by 124Curcumin's Biomedical ApplicationsNeuroinflammation and Neurodegeneration MechanismsCytokine Signaling Pathways and Interactions

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Curcumin Regulates Anti-Inflammatory Responses by JAK/STAT/SOCS Signaling Pathway in BV-2 Microglial Cells

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Porro, Chiara, Cianciulli, Antonia, Trotta, Teresa, Lofrumento, Dario Domenico, Panaro, Maria Antonietta·2019·Biology
DOI 10.3390/biology8030051PMC PMC6784227
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Abstract

Microglia play important physiological roles in central nervous system (CNS) homeostasis and in the pathogenesis of inflammatory brain diseases. Inflammation stimulates microglia to secrete cytokines and chemokines that guide immune cells to sites of injury/inflammation. Neuroinflammation is also strongly implicated in the pathogenesis of a number of neurodegenerative diseases, including Alzheimer's disease and Parkinson's disease, for which nutritional intervention could represent a benefit due to a lack of clinically efficacious drugs. To this end, the anti-inflammatory mechanisms of several phytochemicals, including curcumin, have been extensively studied. The present experiments show that the administration of curcumin is able to increase the production of the anti-inflammatory cytokines, IL-4 and IL-10, in murine BV-2 microglial cells treated with lipopolysaccharide (LPS). Consistent with these data, curcumin stimulation upregulates the expression of Suppressors of cytokine signaling (SOCS)-1, whereas phosphorylation of the JAK2 and STAT3 was reduced. Taken together, these results provide evidence that curcumin is able to regulate neuroinflammatory reactions by eliciting anti-inflammatory responses in microglia through JAK/STAT/SOCS signaling pathway modulation.

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Crossref · 10.3390/biology8030051

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Figure 1 Effect of different concentrations (10, 30, 50 μM) of curcumin (Cur) on anti-inflammatory cytokines production in lipopolysaccharide (LPS)-stimulated BV-2 cells. C = controls. (A) ELISA assay. Cells were treated with the indicated concentrations of curcumin for 1 h before LPS (1 μg/mL) treatment. Supernatants were collected at 24 h after LPS treatment, and the amounts of IL-4 and IL-10 were measured. (B) Reverse transcriptase–polymerase chain reaction (RT-PCR). BV-2 microglial cells were incubated for 6 h with LPS alone or after 1 h of curcumin pretreatment. The experiments were performed in triplicate, and data are expressed as the mean ± SD of five independent experiments. * p < 0.05.
Figure 1 Effect of different concentrations (10, 30, 50 μM) of curcumin (Cur) on anti-inflammatory cytokines production in lipopolysaccharide (LPS)-stimulated BV-2 cells. C = controls. (A) ELISA assay. Cells were treated with the indicated concentrations of curcumin for 1 h before LPS (1 μg/mL) treatment. Supernatants were collected at 24 h after LPS treatment, and the amounts of IL-4 and IL-10 were measured. (B) Reverse transcriptase–polymerase chain reaction (RT-PCR). BV-2 microglial cells were incubated for 6 h with LPS alone or after 1 h of curcumin pretreatment. The experiments were performed in triplicate, and data are expressed as the mean ± SD of five independent experiments. * p < 0.05.CC BY · PubMed Central
Figure 2 Immunoblot analysis of (A) JAK2, (B) STAT3, and (C) SOCS-1 expression. BV-2 microglial cells were treated for 24 h with 1 μg/mL LPS alone or in the presence of various concentrations (10, 30, 50 μM) of curcumin (Cur). Untreated cells were used as controls (C). Bar graphs represent phospho/total JAK2 (A), phospho/total STAT3 (B) and SOCS-1 (C). Protein expression levels were normalized to β-actin, and results of densitometric analysis are expressed as means ± SD of five independent experiments. * p < 0.05.
Figure 2 Immunoblot analysis of (A) JAK2, (B) STAT3, and (C) SOCS-1 expression. BV-2 microglial cells were treated for 24 h with 1 μg/mL LPS alone or in the presence of various concentrations (10, 30, 50 μM) of curcumin (Cur). Untreated cells were used as controls (C). Bar graphs represent phospho/total JAK2 (A), phospho/total STAT3 (B) and SOCS-1 (C). Protein expression levels were normalized to β-actin, and results of densitometric analysis are expressed as means ± SD of five independent experiments. * p < 0.05.CC BY · PubMed Central
Figure 3 Effects of SOCS-1 silencing on JAK2/STAT3 activation in BV-2 cells. Microglial cells were transfected with either control siRNA (scramble RNA) or SOCS-1 specific siRNA (siSOCS1 RNA). Untransfected cells were used as control (left A and B). (A) RT-PCR analysis of p-JAK2 expression in LPS-activated cells (LPS) in absence or in presence of 50 μM curcumin (50 Cur + LPS). (B) RT-PCR analysis of p-STAT3 expression in LPS-activated cells (LPS) in absence or in presence of 50 μM curcumin (50 Cur + LPS). Results are expressed as means ± SD of five independent experiments. * p < 0.05.
Figure 3 Effects of SOCS-1 silencing on JAK2/STAT3 activation in BV-2 cells. Microglial cells were transfected with either control siRNA (scramble RNA) or SOCS-1 specific siRNA (siSOCS1 RNA). Untransfected cells were used as control (left A and B). (A) RT-PCR analysis of p-JAK2 expression in LPS-activated cells (LPS) in absence or in presence of 50 μM curcumin (50 Cur + LPS). (B) RT-PCR analysis of p-STAT3 expression in LPS-activated cells (LPS) in absence or in presence of 50 μM curcumin (50 Cur + LPS). Results are expressed as means ± SD of five independent experiments. * p < 0.05.CC BY · PubMed Central
Figure 4 Effect of SOCS-1 downregulation on microglial polarization, determined by RT-PCR. Microglial cells were transfected with either control siRNA (scramble RNA) or SOCS-1 specific siRNA (siSOCS1 RNA). Untransfected cells were used as control. BV-2 microglial cells were incubated for 6 h with LPS alone, or after 1 h of pre-treatment with curcumin (50 μM). (A) M1-like (iNOS) expression marker. (B) M2-like (ARG-1) expression marker. Experiments were performed in triplicate, and data are expressed as the mean ± SD of five independent experiments. * p < 0.05.
Figure 4 Effect of SOCS-1 downregulation on microglial polarization, determined by RT-PCR. Microglial cells were transfected with either control siRNA (scramble RNA) or SOCS-1 specific siRNA (siSOCS1 RNA). Untransfected cells were used as control. BV-2 microglial cells were incubated for 6 h with LPS alone, or after 1 h of pre-treatment with curcumin (50 μM). (A) M1-like (iNOS) expression marker. (B) M2-like (ARG-1) expression marker. Experiments were performed in triplicate, and data are expressed as the mean ± SD of five independent experiments. * p < 0.05.CC BY · PubMed Central
Figure 5 Effect of SOCS-1 downregulation on cytokine production determined by RT-PCR. Microglial cells were transfected with either control siRNA (scramble RNA) or SOCS-1 specific siRNA (siSOCS1 RNA). Untransfected cells were used as control (C). (A) Pro-inflammatory cytokine production in LPS-stimulated BV-2 cells. (B) Anti-inflammatory cytokine production in LPS-activated BV-2 microglial cells. Cells were incubated for 6 h with LPS alone, or after 1 h of pretreatment with 50 μM curcumin (Cur). Experiments were performed in triplicate, and data are expressed as the mean ± SD of five independent experiments. * p < 0.05.
Figure 5 Effect of SOCS-1 downregulation on cytokine production determined by RT-PCR. Microglial cells were transfected with either control siRNA (scramble RNA) or SOCS-1 specific siRNA (siSOCS1 RNA). Untransfected cells were used as control (C). (A) Pro-inflammatory cytokine production in LPS-stimulated BV-2 cells. (B) Anti-inflammatory cytokine production in LPS-activated BV-2 microglial cells. Cells were incubated for 6 h with LPS alone, or after 1 h of pretreatment with 50 μM curcumin (Cur). Experiments were performed in triplicate, and data are expressed as the mean ± SD of five independent experiments. * p < 0.05.CC BY · PubMed Central

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Metadata

Authors
Porro, C., Cianciulli, A., Trotta, T., Lofrumento, D. D., & Panaro, M. A.
Affiliations
  • University of Foggia (IT)
  • University of Bari Aldo Moro (IT)
  • University of Salento (IT)
Corresponding author
Panaro, Maria Antonietta
ORCID
Venue
Biology
Publisher
MDPI AG
Published
2019-06-27
Volume
8
Issue
3
Pages
51
ISSN
2079-7737 (ISSN-L)
DOI
10.3390/biology8030051
PMC
PMC6784227
OpenAlex
W2953720445
Cited by
124
References
40
Citations by year
2026: 10 · 2025: 14 · 2024: 19 · 2023: 28 · 2022: 17 · 2021: 18 · 2020: 15 · 2019: 3
FWCI
4.58
Type
journal article
Language
en
Open access
Licence
Locations
  • Biology (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Institutional Research Information System University of Ferrara (University of Ferrara) (submittedVersion, public-domain)
  • AIRUS (submittedVersion)
  • CINECA IRIS Institutional Research Information System (University of Bari Aldo Moro) (submittedVersion, cc-by-nc-nd)
Funders
Università degli Studi di Foggia
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Curcumin's Biomedical Applications
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Life Sciences › Immunology and Microbiology › Immunology › Cytokine Signaling Pathways and Interactions
Keywords
curcumin, microglia, neuroinflammation, SOCS1, STAT3 phosphorylation, JAK2 phosphorylation, IL-4, IL-10, anti-inflammatory cytokines, lipopolysaccharide, BV-2 cells, neurodegenerative diseases, phytochemicals

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