甲炎康泰对自身免疫性甲状腺炎大鼠PI3K/Akt/mTOR信号通路的影响

Effects of Jiayan Kangtai on PI3K/Akt/mTOR signaling pathway in autoimmune thyroiditis rats

目的观察甲炎康泰对自身免疫性甲状腺炎(autoimmune thyroiditis,AIT)大鼠磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)/哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路的影响。方法皮下注射猪甲状腺球蛋白联合饮用高碘水建立AIT大鼠模型,将造模成功的AIT大鼠随机分为模型组及甲炎康泰组,每组10只,另设健康Lewis大鼠10只为正常组。甲炎康泰组大鼠给予甲炎康泰水溶液灌胃,其余组予等体积去离子水灌胃,灌药体积为1 mL/100 mg,每日一次。连续给药8周。酶联免疫吸附剂测定法检测大鼠血清中甲状腺过氧化物酶抗体(thyroid peroxidase antibody,TPOAb)滴度;取甲状腺组织进行苏木精—伊红(hematoxylin-eosin,HE)染色,观察病理变化;实时荧光定量PCR检测甲状腺组织中PI3K、Akt、mTOR mRNA的表达;免疫组化检测甲状腺组织中PI3K、P-Akt、P-mTOR的表达情况。结果经甲炎康泰灌胃给药干预8周后,与模型组相比,甲炎康泰组TPOAb滴度值显著降低(P<0.05),淋巴细胞浸润减轻,PI3K、Akt和mTOR mRNA表达降低(均P<0.05),PI3K、P-Akt、P-mTOR蛋白表达量降低(均P<0.05)。结论甲炎康泰可能通过抑制PI3K/Akt/mTOR信号通路,促进细胞自噬,维持T细胞稳态,进而减轻炎症反应,延缓AIT发生发展。

Objective To observe the effect of Jiayan Kangtai on the signal pathway of phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian Sirolimus target(mTOR)in autoimmune thyroiditis(AIT)rats.Methods AIT rat models were established by subcutaneous injection of thyroglobulin combined with drinking high iodine water.AIT rats were randomly divided into the model group and the Jiayan Kangtai group with 10 rats in each group and 10 healthy Lewis rats as the normal group.The rats in the Jiayan Kangtai group were given Jiayan Kangtai water solution and the rats in other groups were given the same volume of deionized water once a day for 8 weeks.The titer of Thyroid peroxidase antibody(TPOAB)in the serum was detected by enzyme-linked immunosorbent assay(Elisa).The thyroid tissues were stained with eosin(HE)and the pathological changes were observed,the expressions of PI3K,AKT,mTOR mRNA in thyroid tissues were detected by real-time fluorescent quantitative PCR.The expressions of PI3K,P-Akt,P-mTOR in thyroid tissues were detected by immunohistochemistry.Results After 8 weeks of intervention,the titer of TPOAB,lymphocyte infiltration,PI3K,AKT and mTOR mRNA(P<0.05)were significantly lower in the Jiayan Kangtai group than those in the model group(P<0.05),PI3K,P-AKT,P-mTOR protein expression were decreased(P<0.05).Conclusion Jiayan Kangtai may inhibit PI3K/AKT/mTOR signaling pathway,promote autophagy,maintain T cell homeostasis,and then reduce the inflammatory reaction and delay the development of AIT.