逍遥散加味通过miRNA326靶向调节ADAM17对自身免疫性甲状腺炎肝郁脾虚大鼠的抗炎机制研究  

Anti-inflammatory mechanism of Xiaoyaosan on autoimmune thyroiditis rats with liver-qi stagnation and spleendeficiency through miRNA326 targeted regulation of ADAM17

作　　者：李祎楠 沈静雪[1] 史哲[2] 徐和平 冯颖岚 马业明 马小芹 马国强 裴海燕 张文彦 罗川 热娜古力·阿布力孜 刘静 洪欣 LI Yinan;SHEN jingxue;SHI Zhe(Second Department of Endocrinology,Affiliated Central Hospital of Shenyang Medical College,Shenyang Liaoning 110000,China;Shenyang Traditional Chinese Medicine Hospital,Shenyang Liaoning 110000,China)

机构地区：[1]沈阳医学院附属中心医院内分泌二科,辽宁沈阳110000 [2]沈阳市中医院,辽宁沈阳110000 [3]塔城市人民医院援疆专家管理办,新疆塔城834700 [4]塔城市人民医院内一科,新疆塔城834700 [5]塔城市塔城地区人民医院检验科,新疆塔城834700 [6]塔城市人民医院检验科,新疆塔城8347000 [7]塔城市人民医院中医科,新疆塔城834700

出　　处：《四川中医》2023年第11期67-74,共8页Journal of Sichuan of Traditional Chinese Medicine

基　　金：辽宁省自然科学基金计划(科技援疆援藏医疗专项计划);沈阳市卫健委课题计划项目(编号:2022064)。

摘　　要：目的:考察逍遥散加味通过小分子核糖核酸326(miR326)靶向调节肿瘤坏死因子转化酶17(ADAM17)对自身免疫性甲状腺炎肝郁脾虚大鼠抗炎机制的影响。方法:采用高碘水联合甲状腺球蛋白皮下注射对大鼠进行免疫干预建立自身免疫性甲状腺炎,采用慢性束缚应激+过度疲劳+饮食失节的方法复制大鼠肝郁脾虚证模型。随机将33只大鼠按随机数字表法分为模型组、逍遥散加味组、金水宝片组,另12只作为正常组。连续给药8周后,采用酶联免疫吸附测定法(ELISA)检测血清游离三碘甲状腺原(FT3)、游离甲状腺素(FT4)、超敏促甲状腺激素(TSH)、甲状腺球蛋白抗体(TGAb)、甲状腺过氧化酶抗体(TPOAb)水平;逆转录-聚合酶链反应(RT-PCR)检测甲状腺组织miR-326、白细胞介素-17信使核糖核酸(IL-17mRNA)、白细胞介素-4信使核糖核酸(IL-4mRNA)、干扰素γ核糖核酸(IFN-γmRNA)水平,全自动超微量样品蛋白质表达定量分析(Wes)检测甲状腺组织ADAM17蛋白表达,流式细胞法检测CD4^(+)IFN-γ^(+)T细胞、CD4^(+)IL-4+T细胞和CD4^(+)IL-17^(+)T细胞比例,苏木素-伊红染色(HE)染色法检测各组大鼠甲状腺组织病理形态。结果:与模型组比较,逍遥散组大鼠血清TSH水平升高(P<0.01),TGAb和TPOAb水平降低(P<0.01),CD4^(+)IFN-γ^(+)T细胞、CD4^(+)IL-17^(+)T细胞比例降低(P<0.01);甲状腺组织miR326、IL-17 mRNA水平降低(P<0.01);ADAM17蛋白表达升高(P<0.01),甲状腺组织病理学明显改变。结论:逍遥散加味可能通过下调肝郁脾虚型实验性身免疫性甲状腺炎大鼠(EAT)甲状腺组织中的miR-326的表达,从而上调靶蛋白ADAM17的表达,抑制Th17细胞分化,进一步减少IL-17 mRNA的表达,最终产生抗炎作用。Objective:To investigate the effect of modified Xiaoyao Powder on the anti-inflammatory mechanism of autoimmune thyroiditis rats with liver depression and spleen deficiency by targeting ADAM17 through miR326.Methods:The rats were immunized with high iodine water combined with thyroglobulin subcutaneously to establish autoimmune thyroiditis,and the rat model of liver depression and spleen deficiency syndrome was reproduced by the method of chronic restraint stress+excessive fatigue+eating disorder.Thirty three rats were randomly divided into model group,modified Xiaoyao powder group,Jinshuibao tablet group,and the other 12 rats were taken as normal group.After continuous administration for 8 weeks,serum FT3,FT4,TSH,TGAb,TPOAb levels were detected by ELISA;Detection of miR-326,IL-17 mRNA,IL-4 mRNA and IFN in thyroid tissue by PCR-γMRNA level,Wes automatic ultramicro sample protein expression quantitative analysis to detect ADAM17 protein expression in thyroid tissue,flow cytometry to detect CD4^(+)IFN-γ^(+)The proportion of T cells,CD4^(+)IL-4^(+)T cells and CD4^(+)IL-17^(+)T cells was detected by HE staining.Results:Compared with the model group,the serum TSH level of rats in Xiaoyao Powder group increased(P<0.01),TGAb and TPOAb levels decreased(P<0.01),CD4^(+)IFN-γ^(+)The proportion of T cells and CD4^(+)IL-17^(+)T cells decreased(P<0.01);The mRNA levels of miR326 and IL-17 in thyroid tissue decreased(P<0.01);The expression of ADAM17 protein increased(P<0.01),and thyroid histopathology changed significantly.Conclusion:Modified Xiaoyao Powder may down regulate the expression of miR-326 in thyroid tissue of EAT rats,thus up regulate the expression of target protein ADAM17,inhibit the differentiation of Th17 cells,further reduce the expression of IL-17 mRNA,and finally produce anti-inflammatory effect.

关 键 词：甲状腺炎 自身免疫性 逍遥散加味 肝郁脾虚 miRNA326 ADAM17 

分 类 号：R285.5[医药卫生—中药学]