补中益气汤对自身免疫性甲状腺炎小鼠Fas/FADD/Caspase-8细胞凋亡信号通路的影响  

Effect of Buzhong Yiqitang on Fas/FADD/Caspase-8 Cell Apoptotic Signaling Pathway in Mice with Autoimmune Thyroiditis

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27-34,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:探讨补中益气汤对自身免疫性甲状腺炎(AIT)模型小鼠肿瘤坏死因子受体超家族成员6(Fas)/Fas相关死亡结构域蛋白(FADD)/胱天蛋白酶-8(Caspase-8)细胞凋亡信号通路的影响。方法:SPF级8周龄NOD.H-2h4小鼠120只,其中100只饲以高碘水(0.05%NaI)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(硒酵母片)组(3.033×10^(-5) g·kg^(-1)),每组20只,另设20只空白组小鼠。依据分组连续给药8周后取材,原位末端标记法(TUNEL)检测小鼠甲状腺细胞凋亡情况;实时荧光定量聚合酶链式反应(Real-time PCR)检测甲状腺组织中Fas、FADD、Caspase-8、Caspase-3 mRNA表达水平;蛋白免疫印迹法(Western blot)检测甲状腺组织Fas、FADD、Caspase-8、胱天蛋白酶-3(Caspase-3)、活化的(cleaved)Caspase-8、cleaved Caspase-3蛋白的表达并应用免疫组织化学染色法检测甲状腺组织Fas、Caspase-3蛋白定位表达情况。结果:与空白组比较,模型组小鼠荧光显微镜下凋亡细胞阳性表达较多,Fas、FADD、Caspase-8、Caspase-3、cleaved Caspase-8、cleaved Caspase-3的表达明显升高(P<0.05,P<0.01);与模型组比较,各给药组小鼠荧光显微镜下甲状腺凋亡细胞阳性表达减少,Fas、FADD、Caspase-8、Caspase-3、cleaved Caspase-8、cleaved Caspase-3的表达明显降低(P<0.05,P<0.01)。结论:补中益气汤可有效改善AIT小鼠甲状腺细胞凋亡,改善炎性损伤,其机制可能与抑制Fas/FADD/Caspase-8信号通路有关。Objective To investigate the effects of Buzhong Yiqitang on the tumor necrosis factor receptor superfamily member 6(Fas)/Fas related death domain protein(FADD)/Caspase-8 cell apoptotic signaling pathway in mice model with autoimmune thyroiditis(AIT).Method There were 120 SPF grade NOD.H-2h4 mice aged 8 weeks,100 of which were fed with high iodine water(0.05%NaI),and the AIT model was made after 8 weeks.According to random number table method,they were divided into model group,Buzhong Yiqitang low-dose,medium-dose and high-dose groups(4.78,9.56,19.12 g·kg^(-1)),selenium yeast tablet group(3.033×10^(-5) g·kg^(-1)),with 20 mice in each group and 20 control group.The apoptosis of thyroid cells was detected by in situ end labeling(TUNEL)after 8 weeks of administration.The real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression of Fas,FADD,Caspase-8,and Caspase-3 in thyroid tissue.Western blot was used to detect the protein expression of Fas,FADD,Caspase-8,Caspase-3,cleaved Caspase-8,and cleaved Caspase-3 in thyroid tissue,and the immunohistochemical method was used to detect the protein expression of Fas and Caspase-3 in thyroid tissue.Result Compared with control group,there were more positive expressions of apoptotic cells in model group under fluorescence microscope,and the expressions of Fas,FADD,Caspase-8,Caspase-3,cleaved Caspase-8 and cleaved Caspase-3 were significantly increased(P<0.05,P<0.01).Compared with model group,the positive expression of thyroid apoptotic cells in each administration group was decreased under fluorescence microscope,and the expressions of Fas,FADD,Caspase-8,Caspase-3,cleaved Caspase-8,cleaved Caspase-3 were significantly decreased(P<0.05,P<0.01).Conclusion Buzhong Yiqitang can effectively improve thyroid cell apoptosis and inflammatory injury in AIT mice.The mechanism may be related to the inhibition of Fas/FADD/Caspase-8 signaling pathway.

关 键 词：补中益气汤 自身免疫性甲状腺炎 细胞凋亡 肿瘤坏死因子受体超家族成员6(Fas) 

分 类 号：R2-0[医药卫生—中医学] R33R289R593