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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

Ba inhibited the capacity of Th1/Th17 cells to induce adoptive EAE.For adoptive transfer EAE, single-cell suspensions were derived from spleen and lymph nodes of EAE mice at day 10 p.i. MOG (25 μg/ml) plus IL-12 (10 ng/ml) or IL-23 (10 ng/ml) were added to cultures in the presence or absence of Ba (10 μg/ml) for 3 days. The resulting cells were analyzed by flow cytometry under (a) Th1 (MOG35–55 + IL-12) or (b) Th17 (MOG35–55 + IL-23) polarizing conditions. Cultured T cells mentioned above were i.v. injected into naïve female C57BL/6 mice, 8–10 weeks of age, at 3 × 107 Th1 cells per mouse (c) or 1.5 × 107 Th17 cells per mouse (e). Mice were observed daily for EAE severity. At day 18 post injection, CNS MNCs from these mice were harvested and analyzed by flow cytometry. Absolute numbers of IFN-γ+ (d) or IL-17+ (f) cells were calculated by multiplying the total numbers of MNCs and percentage of IFN-γ+ or IL-17+ cells in CD4+ gate. Lumbar spinal cords were isolated for immunohistochemistry. (g) MBP immunohistochemistry on spinal cord sections of mice adoptively transferred Th1/Th17 cells mentioned above. (h) Quantitative analysis of MBP intensity measured at random areas in the white matter of spinal cords using Imagepro. Data are expressed as mean ± SEM (n = 5 each group). *P < 0.05; **P < 0.01; ***P < 0.001. One representative of three experiments is shown.
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f4: Ba inhibited the capacity of Th1/Th17 cells to induce adoptive EAE.For adoptive transfer EAE, single-cell suspensions were derived from spleen and lymph nodes of EAE mice at day 10 p.i. MOG (25 μg/ml) plus IL-12 (10 ng/ml) or IL-23 (10 ng/ml) were added to cultures in the presence or absence of Ba (10 μg/ml) for 3 days. The resulting cells were analyzed by flow cytometry under (a) Th1 (MOG35–55 + IL-12) or (b) Th17 (MOG35–55 + IL-23) polarizing conditions. Cultured T cells mentioned above were i.v. injected into naïve female C57BL/6 mice, 8–10 weeks of age, at 3 × 107 Th1 cells per mouse (c) or 1.5 × 107 Th17 cells per mouse (e). Mice were observed daily for EAE severity. At day 18 post injection, CNS MNCs from these mice were harvested and analyzed by flow cytometry. Absolute numbers of IFN-γ+ (d) or IL-17+ (f) cells were calculated by multiplying the total numbers of MNCs and percentage of IFN-γ+ or IL-17+ cells in CD4+ gate. Lumbar spinal cords were isolated for immunohistochemistry. (g) MBP immunohistochemistry on spinal cord sections of mice adoptively transferred Th1/Th17 cells mentioned above. (h) Quantitative analysis of MBP intensity measured at random areas in the white matter of spinal cords using Imagepro. Data are expressed as mean ± SEM (n = 5 each group). *P < 0.05; **P < 0.01; ***P < 0.001. One representative of three experiments is shown.

Mentions: To test the effect of Ba on the pathogenicity of Th1 and Th17 cells, we prepared single-cell suspensions from spleen and lymph nodes of mice that had been immunized with MOG35–55 seven days earlier and cultured them under Th1 (MOG35–55 +IL-12) or Th17 (MOG35–55 +IL-23) polarizing conditions in the presence or absence of Ba for 3 days. The resulting cells were analyzed for the percentage of T cell subsets, expression of transcription factor, adhesion molecules and activation markers by flow cytometry. Ba treatment significantly reduced numbers of IFN-γ- and IL-17-producting cells under Th1 and Th17 polarizing conditions, respectively (P < 0.001), while the numbers of Th2 (CD4+IL-4+) and Treg (CD4+ Foxp3+) cells did not differ (Fig. 4a,b). Consistent with this observation, expression of key transcription factors T-bet for Th1 cells and ROR-γt for Th17 cells, but not Foxp3 for Treg, was significantly decreased in the presence of Ba under Th1 or Th17 polarizing conditions (P < 0.01, Fig. 4a,b). Surface epitopes such as adhesion molecules (VLA-4 and ICAM-1) and T cell activation markers (CD62LlowCD44hi) were also significantly inhibited under Ba treatment (Fig. 4a,b). These results indicated that Ba inhibits encephalitogenicity of Th cells by suppressing production of pro-inflammatory cytokines, crucial adhesion molecule expression and T cell activation.


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)

Ba inhibited the capacity of Th1/Th17 cells to induce adoptive EAE.For adoptive transfer EAE, single-cell suspensions were derived from spleen and lymph nodes of EAE mice at day 10 p.i. MOG (25 μg/ml) plus IL-12 (10 ng/ml) or IL-23 (10 ng/ml) were added to cultures in the presence or absence of Ba (10 μg/ml) for 3 days. The resulting cells were analyzed by flow cytometry under (a) Th1 (MOG35–55 + IL-12) or (b) Th17 (MOG35–55 + IL-23) polarizing conditions. Cultured T cells mentioned above were i.v. injected into naïve female C57BL/6 mice, 8–10 weeks of age, at 3 × 107 Th1 cells per mouse (c) or 1.5 × 107 Th17 cells per mouse (e). Mice were observed daily for EAE severity. At day 18 post injection, CNS MNCs from these mice were harvested and analyzed by flow cytometry. Absolute numbers of IFN-γ+ (d) or IL-17+ (f) cells were calculated by multiplying the total numbers of MNCs and percentage of IFN-γ+ or IL-17+ cells in CD4+ gate. Lumbar spinal cords were isolated for immunohistochemistry. (g) MBP immunohistochemistry on spinal cord sections of mice adoptively transferred Th1/Th17 cells mentioned above. (h) Quantitative analysis of MBP intensity measured at random areas in the white matter of spinal cords using Imagepro. Data are expressed as mean ± SEM (n = 5 each group). *P < 0.05; **P < 0.01; ***P < 0.001. One representative of three experiments is shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Ba inhibited the capacity of Th1/Th17 cells to induce adoptive EAE.For adoptive transfer EAE, single-cell suspensions were derived from spleen and lymph nodes of EAE mice at day 10 p.i. MOG (25 μg/ml) plus IL-12 (10 ng/ml) or IL-23 (10 ng/ml) were added to cultures in the presence or absence of Ba (10 μg/ml) for 3 days. The resulting cells were analyzed by flow cytometry under (a) Th1 (MOG35–55 + IL-12) or (b) Th17 (MOG35–55 + IL-23) polarizing conditions. Cultured T cells mentioned above were i.v. injected into naïve female C57BL/6 mice, 8–10 weeks of age, at 3 × 107 Th1 cells per mouse (c) or 1.5 × 107 Th17 cells per mouse (e). Mice were observed daily for EAE severity. At day 18 post injection, CNS MNCs from these mice were harvested and analyzed by flow cytometry. Absolute numbers of IFN-γ+ (d) or IL-17+ (f) cells were calculated by multiplying the total numbers of MNCs and percentage of IFN-γ+ or IL-17+ cells in CD4+ gate. Lumbar spinal cords were isolated for immunohistochemistry. (g) MBP immunohistochemistry on spinal cord sections of mice adoptively transferred Th1/Th17 cells mentioned above. (h) Quantitative analysis of MBP intensity measured at random areas in the white matter of spinal cords using Imagepro. Data are expressed as mean ± SEM (n = 5 each group). *P < 0.05; **P < 0.01; ***P < 0.001. One representative of three experiments is shown.
Mentions: To test the effect of Ba on the pathogenicity of Th1 and Th17 cells, we prepared single-cell suspensions from spleen and lymph nodes of mice that had been immunized with MOG35–55 seven days earlier and cultured them under Th1 (MOG35–55 +IL-12) or Th17 (MOG35–55 +IL-23) polarizing conditions in the presence or absence of Ba for 3 days. The resulting cells were analyzed for the percentage of T cell subsets, expression of transcription factor, adhesion molecules and activation markers by flow cytometry. Ba treatment significantly reduced numbers of IFN-γ- and IL-17-producting cells under Th1 and Th17 polarizing conditions, respectively (P < 0.001), while the numbers of Th2 (CD4+IL-4+) and Treg (CD4+ Foxp3+) cells did not differ (Fig. 4a,b). Consistent with this observation, expression of key transcription factors T-bet for Th1 cells and ROR-γt for Th17 cells, but not Foxp3 for Treg, was significantly decreased in the presence of Ba under Th1 or Th17 polarizing conditions (P < 0.01, Fig. 4a,b). Surface epitopes such as adhesion molecules (VLA-4 and ICAM-1) and T cell activation markers (CD62LlowCD44hi) were also significantly inhibited under Ba treatment (Fig. 4a,b). These results indicated that Ba inhibits encephalitogenicity of Th cells by suppressing production of pro-inflammatory cytokines, crucial adhesion molecule expression and T cell activation.

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