黄芩苷在胶原蛋白诱发性关节炎小鼠中抑制BLyS介导的Th17细胞分化和炎症反应  

Baicalin inhibits BLyS-mediated Th17 cell differentiation and inflammatory response in collagen-induced arthritis mice

作　　者：蔡志波[1] 林艳[1] 陈英虎[2] CAI Zhi-bo;LIN Yan;CHEN Ying-hu(Department of Pharmacy,The Children's Hospital of Zhejiang University School of Medicine/National Center for Clinical Medicine of Children's Health and Disease,Hangzhou 310052,China;Department of Infectious Disease,The Children's Hospital of Zhejiang University School of Medicine/National Center for Clinical Medicine of Children's Health and Disease,Hangzhou 310052,China)

机构地区：[1]浙江大学医学院附属儿童医院/国家儿童健康与疾病临床医学研究中心药剂科,浙江杭州310052 [2]浙江大学医学院附属儿童医院/国家儿童健康与疾病临床医学研究中心感染科,浙江杭州310052

出　　处：《中国病理生理杂志》2020年第7期1294-1301,共8页Chinese Journal of Pathophysiology

基　　金：浙江省自然科学基金资助项目(No.LY19H100003)。

摘　　要：目的:探讨黄芩苷对胶原蛋白诱发性关节炎(collagen-induced arthritis,CIA)小鼠的辅助性T细胞17(T helper 17 cells,Th17细胞)分化和炎症反应的调控作用及机制。方法:在第1天,SPF级雄性DBA/1小鼠经背部皮下注射鸡源II型胶原蛋白(chick collagen type II,CII)+完全弗氏佐剂(complete Freund’s adjuvant,CFA)致敏;在第21天,经尾根部皮下注射CII+不完全弗氏佐剂(incomplete Freund’s adjuvant,IFA)强化免疫;在第28~42天分别灌胃50、100和200 mg·kg^-1·d^-1黄芩苷。关节炎指数(arthritis index,AI)积分评估关节炎的严重程度;HE染色观察膝关节病理学改变;ELISA法检测血清中IL-1β、TNF-α、IgG、IgG2a和IL-17A的水平以及关节组织中IL-17A的水平;流式细胞术分析脾脏和肠系膜淋巴结(mesenteric lymph nodes,MLN)中CD4^+IL-17A^+T细胞比例;Western blot检测关节、脾脏和MLN中B淋巴细胞刺激物(B-lymphocyte stimulator,BLyS)的表达。另外,用2μg/L BLyS和20μmol/L黄芩苷处理CD4^+T细胞群48 h后,流式细胞术分析CD4^+IL-17A^+T细胞比例,ELISA法检测上清液中IL-1β和TNF-α的水平。结果:黄芩苷能剂量依赖性地减轻CIA(P<0. 05)。黄芩苷能剂量依赖性地降低CIA小鼠血清中IL-1β、TNF-α、IgG、IgG2a和IL-17A水平和关节组织中IL-17A水平,减少脾脏和MLN中Th17细胞的比例,抑制关节、脾脏和MLN中BLyS的表达(P<0. 05)。另外,黄芩苷能抑制BLyS诱导的CD4^+T细胞群中Th17细胞的分化及IL-1β和TNF-α的分泌(P<0. 05)。结论:黄芩苷能通过降低BLyS介导的Th17细胞分化和炎症反应来发挥抗CIA效应。AIM:To investigate the regulatory effect and mechanism of baicalin on T helper 17(Th17)cell differentiation and inflammatory response in collagen-induced arthritis(CIA)mice.METHODS:Male DBA/1 mice were sensitized by chick collagen type II(CII)+complete Freund’s adjuvant(CFA)via subcutaneous injection into the back on day 1,intensively immunized by CII+incomplete Freund’s adjuvant(IFA)via subcutaneous injection into the tail root on day 21,and finally intragastrically treated with 50,100 and 200 mg·kg^-1·d^-1 baicalin on days 2842. The degree of arthritis was evaluated by arthritis index(AI)scoring. Pathological changes of the knee joint were observed by HE staining.The serum levels of IL-1β,TNF-α,IgG,IgG2 a and IL-17 A,and the levels of IL-17 A in joint tissues were measured by ELISA. The proportion of CD4^+IL-17 A^+T cells in spleen and mesenteric lymph nodes(MLN)was analyzed by flow cytometry. The expression of B-lymphocyte stimulator(BLyS)in joint,spleen and MLN tissues was detected by Western blot.In addition,after CD4^+T cell population were treated with BLyS at 2 μg/L and baicalin at 20 μmol/L for 48 h,the proportion of CD4^+IL-17 A^+T cells was analyzed by flow cytometry,and the levels of IL-1β and TNF-α in supernatant were measured by ELISA.RESULTS:Baicalin attenuated CIA in a dose-dependent manner(P<0. 05). Baicalin decreased the serum levels of IL-1β,TNF-α,IgG,IgG2 a and IL-17 A and the levels of IL-17 A in joint tissues,reduced the proportion of Th17 cells in spleen and MLN tissues,and inhibited the expression of BLyS in joint,spleen and MLN tissues in a dose-dependent manner(P<0. 05). Moreover,baicalin inhibited Th17 cell differentiation and the secretion of IL-1β and TNF-αin CD4^+T cell population induced by BLyS.CONCLUSION:Baicalin reduces BLyS-mediated Th17 cell differentiation and inflammatory response to exert the anti-CIA effect.

关 键 词：黄芩苷 类风湿关节炎 TH17细胞 B淋巴细胞刺激物 

分 类 号：R593.22[医药卫生—内科学] R363.2[医药卫生—临床医学]