摘要
目的：观察益气化痰活血方对自身免疫甲状腺炎（AIT）小鼠甲状腺miR-155/SOCS1/STAT3信号通路的影响,探讨中医补气健脾法干预AIT部分免疫学机制。方法：60只8周龄NOD.H-2h4小鼠随机分为6组。正常组（NG）饲以蒸馏水,模型组（MG）、西药组（硒酵母片,Se G）、益气化痰活血方高剂量组（YG-1,36.40 g·Kg-1·d-1）、益气化痰活血方中剂量组（YG-2,18.20 g·Kg-1·d-1）、益气化痰活血方低剂量组（YG-3,9.10 g·Kg-1·d-1）饲以含0.05%碘化钠水8周成模,再灌胃给药8周。光镜下观察甲状腺细胞形态;RT-PCR法检测小鼠甲状腺miR-155、白细胞介素-17（IL-17）mRNA表达;Western blot法测定小鼠甲状腺细胞因子信号抑制因子（SOCS1）、p-STAT3蛋白表达。结果：（1）与NG组比较,MG组甲状腺淋巴细胞浸润明显;与MG组比较,各给药组甲状腺炎均有不同程度改善。（2）与NG组比较,甲状腺IL-17、miR-155mRNA表达显著上调（P〈0.01）,甲状腺p-STAT3蛋白显著升高（P〈0.01）;与MG组比较,Se G、YG-1、YG-2组甲状腺IL-17mRNA表达均显著下调（P〈0.01）;与Se G组比较,YG-2组IL-17mRNA下调更明显（P〈0.01）;与MG组比较,各给药组miR-155mRNA均显著下调（P〈0.01）,其中Se G、YG-2组无统计学差异（P〉0.05）;各给药组p-STAT3均显著下调（P〈0.01）,与Se G组比较,YG-2组下调更明显（P〈0.05）。（3）与NG组比较,MG组甲状腺SOCS1蛋白明显下降（P〈0.01）,与MG组比较,Se G、YG-2、YG-3组SOCS1蛋白水平显著升高（P〈0.01）,YG-1组SOCS1蛋白上调（P〈0.05）。结论：中医补气健脾法可能通过调控miR-155/SOCS1/STAT3信号通路改善AIT免疫失常。
Objective： Toobserve the effect of Yiqi Huatan Huoxue Fang on the miR-155/SOCS1/STAT3 in autoimmune thyroiditis（ AIT） mice,and investigate the potential immune mechanism of TCM Method of Tonifying Spleen-qi in treatment of AIT. Methods： Sixty 8-week-old NOD. H-2 h4 mice were randomly divided into six groups： normal group（ NG）,model group（ MG）,Selenious yeast group（ Se G）,2 times dose of Yiqi Huatan Huoxue Fanggroup（ YG-1,36. 40 g·Kg-1·d-1）,Yiqi Huatan Huoxue Fang group（ YG-2,18. 20 g·Kg-1·d-1） and times dose of Yiqi Huatan HuoxueFanggroup（ YG-3,9. 10 g·Kg-1·d-1）. The mice in NG were fed with distilled water,and the others weresupplied with 0. 05 % Na I for 8 weeks to induce AIT. All mice were sacrificed after 8 weeks of decoction or sterile water gavages.Then thyrocyte morphology was observed under optical microscopy; the expressions of IL-17 mRNA and miR-155 mRNA in mice thyroid were detected by RT-PCRrespectively,and SOCS1 and p-STAT3 protein expressions were determined by Western blot respectively. Results：（1）Compared with the NG,intrathyroidal lymphocyte infiltration of the MG was more obvious,and that of all the medicated groups alleviated when compared with MG.（2）Compared to the NG,and the mRNAexpressions of IL-17,miR-155 and the protein expression of p-STAT3 were all increased markedly in the MG（ P 〈0. 01）; Compared to the MG,the expression of IL-17 mRNA was decreased in the Se G,YG-1 and YG-2 re-markably（ P 〈0. 01）,and that in YG-2 was lower when compared with the Se G（ P〈 0. 01）; Compared to the MG,the expression of miR-155 mRNA was decreased in all the medicated group（ P 0. 01）,and there was no statistic difference in YG-2 and Se G（ P〉 0. 05）; the expression of p-STAT3 was decreased in all the medicated group compared with the MG（ P〈 0. 01）,and compared to the Se G,p-STAT3 expression in YG-2 decreased significantly（ P〈 0. 05）.（3）Compared to the NG,the expression of SOCS1 was decreesed obviously in the MG（ P 〈0. 01）; Compared to the MG,SOCS1 expression in Se G,YG-2 and YG-3 were all increased remarkably（ P〈 0. 01） while that in YG-1 was increased（ P 〈0. 05）. Conclusion： The mechanism of TCM Method of Tonifying Spleen-qi on AIT mice with immune injury may be related to regulate miR-155/SOCS1/STAT3 signaling pathway.
作者
杨潇
宋囡
周喜玉
曹慧敏
王智民
徐佳
高天舒
YANG Xiao;SONG Nan;ZHOU Xiyu;CAO Huimin;WANG Zhimin;XU Jia;GAO Tianshu(Second Affiliated Hospital of Liaoning University of Chinese Medicine,Shenyang 110034,Liaoning,China;Liaoning University of Chinese Medicine,Shenyang 110032,Liaoning,China;Affiliated Hospital of Liaoning University of Chinese Medicine,Shenyang 110032,Liaoning,China)
出处
《中华中医药学刊》
CAS
北大核心
2018年第9期2074-2077,I0003,共5页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金青年科学基金项目(81503474)
辽宁省教育厅一般项目(L2015347)
辽宁省科技厅博士启动基金项目(201501110)
辽宁中医药大学大学生创新创业训练计划项目(201510162000011)
作者简介
杨潇（1981-），女，辽宁大连人，副主任医师、副教授，硕士研究生导师，博士，研究方向：中医药防治自身免疫性甲状腺炎机制。;通讯作者：高天舒（1967-），男，辽宁抚顺人，主任医师、教授，博士研究生导师，博士，研究方向：中医药防治甲状腺疾病。E-mail：gaotianshu67@163．com。
