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Therapeutic effect of baicalin on experimental autoimmune encephalomyelitis is mediated by SOCS3 regulatory pathway.

Zhang Y, Li X, Ciric B, Ma CG, Gran B, Rostami A, Zhang GX - Sci Rep (2015)

Bottom Line: Natural compounds derived from medicinal plants have long been considered a rich source of novel therapeutic agents.Ba treatment effectively ameliorated clinical disease severity in myelin oligodendrocyte glycoprotein (MOG)35-55 peptide-induced EAE, and reduced inflammation and demyelination of the central nervous system (CNS).Taken together, our findings demonstrate that Ba has significant potential as a novel anti-inflammatory agent for therapy of autoimmune diseases such as MS.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Thomas Jefferson University, Philadelphia, PA, USA.

ABSTRACT
Natural compounds derived from medicinal plants have long been considered a rich source of novel therapeutic agents. Baicalin (Ba) is a bioactive flavonoid compound derived from the root of Scutellaria baicalensis, an herb widely used in traditional medicine for the treatment of various inflammatory diseases. In this study, we investigate the effects and mechanism of action of Ba in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). Ba treatment effectively ameliorated clinical disease severity in myelin oligodendrocyte glycoprotein (MOG)35-55 peptide-induced EAE, and reduced inflammation and demyelination of the central nervous system (CNS). Ba reduced infiltration of immune cells into the CNS, inhibited expression of proinflammatory molecules and chemokines, and prevented Th1 and Th17 cell differentiation via STAT/NFκB signaling pathways. Further, we showed that SOCS3 induction is essential to the effects of Ba, given that the inhibitory effect of Ba on pathogenic Th17 responses was largely abolished when SOCS3 signaling was knocked down. Taken together, our findings demonstrate that Ba has significant potential as a novel anti-inflammatory agent for therapy of autoimmune diseases such as MS.

No MeSH data available.


Related in: MedlinePlus

Th1 and Th17 cell subsets are selectively reduced by Ba via STAT and NF-κB signaling pathways.EAE mice were treated with Ba or PBS starting at day 10 p.i. and splenocytes or CNS MNCs from these mice were harvested at day 18 p.i. (a) Subsets of Th1, Th17, Th2, and Treg cells in CD4+ gate were analyzed by intracellular staining of IFN-γ, IL-17, IL-4, and Foxp3, following stimulation with MOG35–55 (25 μg/ml) for 72 h for spleen or with MOG35–55 (10 μg/ml) for 24 h for CNS cells. (b) Percentages of cells positive for these cytokines in CNS (up) and spleen (down) are expressed as mean ± SEM (n = 3 each group). (c) Supernatants derived from splenocyte cultures described in (a) were analyzed for the level of indicated cytokines (mean ± SEM; n = 6 each group). (d) mRNA levels of T-bet, RORγt, GATA3, and Foxp3 from spleens of EAE mice treated with Ba or PBS were analyzed by real-time PCR. (e) Splenocytes were cultured in the presence of MOG35–55 (25 μg/ml) for 72 h and analyzed by intracellular staining for phosphorylation level of indicated STAT proteins. (f) Supernatants from cell cultures in (c) were analyzed for phosphorylated IκBα, NF-κB p65, and phosphorylated NF-κB p65 by PathScan Inflammation Multi-Target Sandwich ELISA Kit (n = 6 each group). Data are expressed as mean ± SEM. #, *P < 0.05; **P < 0.01; ***P < 0.001. One representative of three experiments is shown.
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f3: Th1 and Th17 cell subsets are selectively reduced by Ba via STAT and NF-κB signaling pathways.EAE mice were treated with Ba or PBS starting at day 10 p.i. and splenocytes or CNS MNCs from these mice were harvested at day 18 p.i. (a) Subsets of Th1, Th17, Th2, and Treg cells in CD4+ gate were analyzed by intracellular staining of IFN-γ, IL-17, IL-4, and Foxp3, following stimulation with MOG35–55 (25 μg/ml) for 72 h for spleen or with MOG35–55 (10 μg/ml) for 24 h for CNS cells. (b) Percentages of cells positive for these cytokines in CNS (up) and spleen (down) are expressed as mean ± SEM (n = 3 each group). (c) Supernatants derived from splenocyte cultures described in (a) were analyzed for the level of indicated cytokines (mean ± SEM; n = 6 each group). (d) mRNA levels of T-bet, RORγt, GATA3, and Foxp3 from spleens of EAE mice treated with Ba or PBS were analyzed by real-time PCR. (e) Splenocytes were cultured in the presence of MOG35–55 (25 μg/ml) for 72 h and analyzed by intracellular staining for phosphorylation level of indicated STAT proteins. (f) Supernatants from cell cultures in (c) were analyzed for phosphorylated IκBα, NF-κB p65, and phosphorylated NF-κB p65 by PathScan Inflammation Multi-Target Sandwich ELISA Kit (n = 6 each group). Data are expressed as mean ± SEM. #, *P < 0.05; **P < 0.01; ***P < 0.001. One representative of three experiments is shown.

Mentions: The decreased numbers of CD4+ T cells in the CNS of mice treated with Ba prompted us to investigate which subsets among these cells were affected. Ba significantly reduced the percentages of MOG-reactive Th1 (CD4+IFN-γ+) and Th17 (CD4+ IL-17+) cells, both in the spleen (P = 0.0005 and P = 0.0067, respectively) and the CNS (P = 0.0028 and P = 0.0006, respectively) compared with the vehicle control, while numbers of Th2 (CD4+IL4+) and Treg (CD4+Foxp3+) cells did not differ (Fig. 3a,b). Consistent with this, production of IFN-γ and IL-17 was significantly reduced by Ba treatment (P < 0.001, Fig. 3c). We also showed that GM-CSF, a critical cytokine in Th17 cell pathogenicity24, was significantly decreased by Ba treatment (P < 0.001, Fig. 3c). In contrast, low but detectable levels of the Th2 cytokines IL-4 and IL-5 were not affected by Ba (Fig. 3c). Anti-inflammatory cytokine IL-10 showed only a small, but significant, increase (Fig. 3c). We also defined the effect of Ba on cytokine production of splenocytes from untreated EAE mice in vitro. Ba significantly suppressed IFN-γ, IL-17 and GM-CSF, and up-regulated IL-10 production in vitro (Fig. s3).


Therapeutic effect of baicalin on experimental autoimmune encephalomyelitis is mediated by SOCS3 regulatory pathway.

Zhang Y, Li X, Ciric B, Ma CG, Gran B, Rostami A, Zhang GX - Sci Rep (2015)

Th1 and Th17 cell subsets are selectively reduced by Ba via STAT and NF-κB signaling pathways.EAE mice were treated with Ba or PBS starting at day 10 p.i. and splenocytes or CNS MNCs from these mice were harvested at day 18 p.i. (a) Subsets of Th1, Th17, Th2, and Treg cells in CD4+ gate were analyzed by intracellular staining of IFN-γ, IL-17, IL-4, and Foxp3, following stimulation with MOG35–55 (25 μg/ml) for 72 h for spleen or with MOG35–55 (10 μg/ml) for 24 h for CNS cells. (b) Percentages of cells positive for these cytokines in CNS (up) and spleen (down) are expressed as mean ± SEM (n = 3 each group). (c) Supernatants derived from splenocyte cultures described in (a) were analyzed for the level of indicated cytokines (mean ± SEM; n = 6 each group). (d) mRNA levels of T-bet, RORγt, GATA3, and Foxp3 from spleens of EAE mice treated with Ba or PBS were analyzed by real-time PCR. (e) Splenocytes were cultured in the presence of MOG35–55 (25 μg/ml) for 72 h and analyzed by intracellular staining for phosphorylation level of indicated STAT proteins. (f) Supernatants from cell cultures in (c) were analyzed for phosphorylated IκBα, NF-κB p65, and phosphorylated NF-κB p65 by PathScan Inflammation Multi-Target Sandwich ELISA Kit (n = 6 each group). Data are expressed as mean ± SEM. #, *P < 0.05; **P < 0.01; ***P < 0.001. One representative of three experiments is shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4663791&req=5

f3: Th1 and Th17 cell subsets are selectively reduced by Ba via STAT and NF-κB signaling pathways.EAE mice were treated with Ba or PBS starting at day 10 p.i. and splenocytes or CNS MNCs from these mice were harvested at day 18 p.i. (a) Subsets of Th1, Th17, Th2, and Treg cells in CD4+ gate were analyzed by intracellular staining of IFN-γ, IL-17, IL-4, and Foxp3, following stimulation with MOG35–55 (25 μg/ml) for 72 h for spleen or with MOG35–55 (10 μg/ml) for 24 h for CNS cells. (b) Percentages of cells positive for these cytokines in CNS (up) and spleen (down) are expressed as mean ± SEM (n = 3 each group). (c) Supernatants derived from splenocyte cultures described in (a) were analyzed for the level of indicated cytokines (mean ± SEM; n = 6 each group). (d) mRNA levels of T-bet, RORγt, GATA3, and Foxp3 from spleens of EAE mice treated with Ba or PBS were analyzed by real-time PCR. (e) Splenocytes were cultured in the presence of MOG35–55 (25 μg/ml) for 72 h and analyzed by intracellular staining for phosphorylation level of indicated STAT proteins. (f) Supernatants from cell cultures in (c) were analyzed for phosphorylated IκBα, NF-κB p65, and phosphorylated NF-κB p65 by PathScan Inflammation Multi-Target Sandwich ELISA Kit (n = 6 each group). Data are expressed as mean ± SEM. #, *P < 0.05; **P < 0.01; ***P < 0.001. One representative of three experiments is shown.
Mentions: The decreased numbers of CD4+ T cells in the CNS of mice treated with Ba prompted us to investigate which subsets among these cells were affected. Ba significantly reduced the percentages of MOG-reactive Th1 (CD4+IFN-γ+) and Th17 (CD4+ IL-17+) cells, both in the spleen (P = 0.0005 and P = 0.0067, respectively) and the CNS (P = 0.0028 and P = 0.0006, respectively) compared with the vehicle control, while numbers of Th2 (CD4+IL4+) and Treg (CD4+Foxp3+) cells did not differ (Fig. 3a,b). Consistent with this, production of IFN-γ and IL-17 was significantly reduced by Ba treatment (P < 0.001, Fig. 3c). We also showed that GM-CSF, a critical cytokine in Th17 cell pathogenicity24, was significantly decreased by Ba treatment (P < 0.001, Fig. 3c). In contrast, low but detectable levels of the Th2 cytokines IL-4 and IL-5 were not affected by Ba (Fig. 3c). Anti-inflammatory cytokine IL-10 showed only a small, but significant, increase (Fig. 3c). We also defined the effect of Ba on cytokine production of splenocytes from untreated EAE mice in vitro. Ba significantly suppressed IFN-γ, IL-17 and GM-CSF, and up-regulated IL-10 production in vitro (Fig. s3).

Bottom Line: Natural compounds derived from medicinal plants have long been considered a rich source of novel therapeutic agents.Ba treatment effectively ameliorated clinical disease severity in myelin oligodendrocyte glycoprotein (MOG)35-55 peptide-induced EAE, and reduced inflammation and demyelination of the central nervous system (CNS).Taken together, our findings demonstrate that Ba has significant potential as a novel anti-inflammatory agent for therapy of autoimmune diseases such as MS.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Thomas Jefferson University, Philadelphia, PA, USA.

ABSTRACT
Natural compounds derived from medicinal plants have long been considered a rich source of novel therapeutic agents. Baicalin (Ba) is a bioactive flavonoid compound derived from the root of Scutellaria baicalensis, an herb widely used in traditional medicine for the treatment of various inflammatory diseases. In this study, we investigate the effects and mechanism of action of Ba in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). Ba treatment effectively ameliorated clinical disease severity in myelin oligodendrocyte glycoprotein (MOG)35-55 peptide-induced EAE, and reduced inflammation and demyelination of the central nervous system (CNS). Ba reduced infiltration of immune cells into the CNS, inhibited expression of proinflammatory molecules and chemokines, and prevented Th1 and Th17 cell differentiation via STAT/NFκB signaling pathways. Further, we showed that SOCS3 induction is essential to the effects of Ba, given that the inhibitory effect of Ba on pathogenic Th17 responses was largely abolished when SOCS3 signaling was knocked down. Taken together, our findings demonstrate that Ba has significant potential as a novel anti-inflammatory agent for therapy of autoimmune diseases such as MS.

No MeSH data available.


Related in: MedlinePlus