补中益气汤通过miR-155/Ndfip1/Pten轴调控Th17细胞改善自身免疫性甲状腺炎的作用机制  

Mechanism of Buzhong Yiqitang in Improving Autoimmune Thyroiditis by Regulating Th17 Cells Through miR-155/Ndfip1/Pten Axis

作　　者：李晓晖 赵卓 陈怡然[1] 曹慧敏[1] 陈丝 王智民[2] 高天舒[2] 刘子玉 杨潇[3] LI Xiaohui;ZHAO Zhuo;CHEN Yiran;CAO Huimin;CHEN Si;WANG Zhimin;GAO Tianshu;LIU Ziyu;YANG Xiao(Liaoning University of Traditional Chinese Medicine(TCM),Shenyang 110847,China;Affiliated Hospital of Liaoning University of TCM,Shenyang110032,China;The Second Affiliated Hospital of Liaoning University of TCM,Shenyang 110034,China;Liaoning Institute of TCM,Shenyang 110034,China)

机构地区：[1]辽宁中医药大学,沈阳110847 [2]辽宁中医药大学附属医院,沈阳110032 [3]辽宁中医药大学附属第二医院,沈阳110034 [4]辽宁省中医药研究院,沈阳110034

出　　处：《中国实验方剂学杂志》2024年第18期19-26,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82274455,82104805);辽宁省教育厅重点攻关项目(JYTZD2023196);辽宁省科技厅联合基金项目(2023-MSLH-154);尹远平全国名老中医药专家传承工作室项目(202205)。

摘　　要：目的:探讨补中益气汤通过微小RNA-155(miR-155)/Nedd4家族相互作用蛋白1(Ndfip1)/磷酸酯酶与张力蛋白同源物(Pten)轴调控辅助性T细胞17(Th17)改善自身免疫性甲状腺炎(AIT)的作用机制。方法:将100只SPF级8周龄NOD.H-2h4小鼠饲以高碘水(0.05%碘化钠)喂养,8周后制成AIT模型,按随机数字表法分为模型组、补中益气汤低、中、高剂量组(4.78、9.56、19.12 g·kg^(-1))、西药组(硒酵母片3.033×10^(-5) g·kg^(-1)),每组20只,另设20只空白组小鼠,空白组及模型组灌胃等体积蒸馏水。连续给药8周后,实时荧光定量聚合酶链式反应(Real-time PCR)检测小鼠甲状腺组织miR-155-5p、Ndfip1、Pten、蛋白质酪氨酸激酶1(Jak1)、信号传导和转录激活因子3(Stat3)、维甲酸相关孤儿受体γt(RORγt)、白细胞介素-17(IL-17)mRNA的表达;蛋白免疫印迹法(Western blot)检测小鼠甲状腺组织Ndfip1、Pten、Jak1、Stat3、RORγt、IL-17蛋白的表达;免疫组化法检测小鼠甲状腺组织中Ndfip1、Pten蛋白表达;流式细胞术检测小鼠脾脏Th17细胞比例。结果:与空白组比较,模型组小鼠脾脏Th17细胞比例显著升高(P<0.01),miR-155-5p、Jak1、Stat3、RORγt、IL-17的表达显著升高(P<0.01),Ndfip1、Pten的表达显著降低(P<0.01);与模型组比较,各给药组小鼠脾脏Th17细胞比例降低(P<0.05,P<0.01),miR-155-5p、Jak1、Stat3、RORγt、IL-17的表达明显减少(P<0.05,P<0.01),Ndfip1、Pten的表达升高(P<0.05,P<0.01)。结论:补中益气汤可改善AIT小鼠自身免疫失常,其机制可能与通过miR-155调控Ndfip1/Pten轴进而调控Th17细胞分化有关。Objective To explore the mechanism of Buzhong Yiqitang in improving autoimmune thyroiditis(AIT)by regulating helper T cell 17(Th17)cells through microRNA-155(miR-155)/Nedd4 family interaction protein 1(Ndfip1)/phosphatase and tensin homology(Pten)axis.Method The 100 SPF grade 8 week-old NOD.H-2h4 mice were fed with high iodine water(0.05%NaI)for 8 weeks,and AIT model was made.They were divided into model group,Buzhong Yiqitang low-,medium-,and high-dose groups(4.78,9.56,19.12 g·kg^(-1)·d^(-1))and selenium yeast tablet group(3.033×10^(-5) g·kg^(-1))according to random number table method.There were 20 mice in each group and 20 mice in the control group.The control group and the model group were given the same amount of distilled water.After 8 weeks of continuous administration,Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the expression of miR-155-5p,Ndfip1,Pten,protein tyrosine kinase 1(Jak1),signaling and transcriptional activator 3(Stat3)retinoic acid-associated orphan receptorγt(RORγt),and interleukin-17(IL-17)mRNA in mouse thyroid tissue.Western blot was used to detect the expression of Ndfip1,Pten,Jak1,Stat3,RORγt,and IL-17 proteins in mouse thyroid tissue,immunohistochemical method was used to detect the expression of Ndfip1 and Pten proteins in mouse thyroid tissue;flow cytometry was used to detect the proportion of Th17 cells in mouse spleen.Result Compared with the control group,the proportion of Th17 cells was increased(P<0.01).The expressions of miR-155-5p,Jak1,Stat3,RORγt and IL-17 were increased(P<0.01),while the expressions of Ndfip1 and Pten were decreased(P<0.01).Compared with model group,the proportion of Th17 cells was decreased(P<0.05,P<0.01).The expressions of miR-155-5p,Jak1,Stat3,RORγt and IL-17 were decreased(P<0.05,P<0.01),while the expressions of Ndfip1 and Pten were increased(P<0.05,P<0.01).Conclusion The application of Buzhong Yiqitang can improve the autoimmune disorder of AIT mice,the mechanism of which may be related to the re

关 键 词：补中益气汤 自身免疫性甲状腺炎(AIT) 辅助性T细胞17(Th17) 微小RNA-155(miR-155) Nedd4家族相互作用蛋白1(Ndfip1) 磷酸酯酶与张力蛋白同源物(Pten) 

分 类 号：R2-0[医药卫生—中医学] R33R289R593R24