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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.


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Role of Ba in Th17 cell differentiation and proliferation.Naïve CD4+ cells were cultured with different concentrations of Ba under the Th17 polarizing condition at 3 days of culture. (a) Percentage of Th17 cells was analyzed by intracellular staining of IL-17. (b) Culture supernatants were analyzed for IL-17 production. (c) IL17a mRNA levels were analyzed by real-time PCR. Intracellular levels of RORγt (d) and phosphorylation of STAT3 (e) were analyzed using flow cytometry. (f) The above-mentioned differentiated Th17 cells were rested, washed and cultured for a second stimulation with IL-23 in the presence of Ba. Percentage of Th17 cells was analyzed by intracellular staining of IL-17. (g) Proliferation of pre-differentiated Th17 cell was measured by BrdU incorporation assay. The same cell preparations as in F were labeled with BrdU and cultured for 24 h. Data are expressed as mean ± SEM. (n = 5 each group). **P < 0.01; ***P < 0.001. One representative of three experiments is shown.
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f6: Role of Ba in Th17 cell differentiation and proliferation.Naïve CD4+ cells were cultured with different concentrations of Ba under the Th17 polarizing condition at 3 days of culture. (a) Percentage of Th17 cells was analyzed by intracellular staining of IL-17. (b) Culture supernatants were analyzed for IL-17 production. (c) IL17a mRNA levels were analyzed by real-time PCR. Intracellular levels of RORγt (d) and phosphorylation of STAT3 (e) were analyzed using flow cytometry. (f) The above-mentioned differentiated Th17 cells were rested, washed and cultured for a second stimulation with IL-23 in the presence of Ba. Percentage of Th17 cells was analyzed by intracellular staining of IL-17. (g) Proliferation of pre-differentiated Th17 cell was measured by BrdU incorporation assay. The same cell preparations as in F were labeled with BrdU and cultured for 24 h. Data are expressed as mean ± SEM. (n = 5 each group). **P < 0.01; ***P < 0.001. One representative of three experiments is shown.

Mentions: Ba also significantly inhibited Th17 cell differentiation (Fig. 6a) and reduced IL-17 A production and expression in a dose-dependent manner (5–20 μg/ml) (Fig. 6b,c). RORγt is the master regulator for Th17 differentiation, and STAT3 is the key transducer of IL-6, IL-21, and IL-23 signaling2829. Consistent with its effect on Th17 differentiation, Ba suppressed RORγt expression (Fig. 6d) as well as its upstream event, STAT3 phosphorylation (Fig. 6e), under Th17 polarizing condition. Further, we found that addition of Ba into already differentiated Th17 cells reduced the percentage and proliferation of CD4+ IL-17+ cells in a dose-dependent manner (Fig. 6f,g). These results, together with those shown in Fig. 5, indicate that Ba suppresses the differentiation and proliferation of both Th1 and Th17 cells.


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)

Role of Ba in Th17 cell differentiation and proliferation.Naïve CD4+ cells were cultured with different concentrations of Ba under the Th17 polarizing condition at 3 days of culture. (a) Percentage of Th17 cells was analyzed by intracellular staining of IL-17. (b) Culture supernatants were analyzed for IL-17 production. (c) IL17a mRNA levels were analyzed by real-time PCR. Intracellular levels of RORγt (d) and phosphorylation of STAT3 (e) were analyzed using flow cytometry. (f) The above-mentioned differentiated Th17 cells were rested, washed and cultured for a second stimulation with IL-23 in the presence of Ba. Percentage of Th17 cells was analyzed by intracellular staining of IL-17. (g) Proliferation of pre-differentiated Th17 cell was measured by BrdU incorporation assay. The same cell preparations as in F were labeled with BrdU and cultured for 24 h. Data are expressed as mean ± SEM. (n = 5 each group). **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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getmorefigures.php?uid=PMC4663791&req=5

f6: Role of Ba in Th17 cell differentiation and proliferation.Naïve CD4+ cells were cultured with different concentrations of Ba under the Th17 polarizing condition at 3 days of culture. (a) Percentage of Th17 cells was analyzed by intracellular staining of IL-17. (b) Culture supernatants were analyzed for IL-17 production. (c) IL17a mRNA levels were analyzed by real-time PCR. Intracellular levels of RORγt (d) and phosphorylation of STAT3 (e) were analyzed using flow cytometry. (f) The above-mentioned differentiated Th17 cells were rested, washed and cultured for a second stimulation with IL-23 in the presence of Ba. Percentage of Th17 cells was analyzed by intracellular staining of IL-17. (g) Proliferation of pre-differentiated Th17 cell was measured by BrdU incorporation assay. The same cell preparations as in F were labeled with BrdU and cultured for 24 h. Data are expressed as mean ± SEM. (n = 5 each group). **P < 0.01; ***P < 0.001. One representative of three experiments is shown.
Mentions: Ba also significantly inhibited Th17 cell differentiation (Fig. 6a) and reduced IL-17 A production and expression in a dose-dependent manner (5–20 μg/ml) (Fig. 6b,c). RORγt is the master regulator for Th17 differentiation, and STAT3 is the key transducer of IL-6, IL-21, and IL-23 signaling2829. Consistent with its effect on Th17 differentiation, Ba suppressed RORγt expression (Fig. 6d) as well as its upstream event, STAT3 phosphorylation (Fig. 6e), under Th17 polarizing condition. Further, we found that addition of Ba into already differentiated Th17 cells reduced the percentage and proliferation of CD4+ IL-17+ cells in a dose-dependent manner (Fig. 6f,g). These results, together with those shown in Fig. 5, indicate that Ba suppresses the differentiation and proliferation of both Th1 and Th17 cells.

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