夏枯草对自身免疫性甲状腺炎大鼠Th相关细胞因子表达及JNK/p38 MAPK信号通路的影响

Effects of Prunella vulgaris on expression of Th cell-related cytokine and JNK/p38 MAPK signaling pathway in rats with autoimmune thyroiditis

目的研究夏枯草对自身免疫性甲状腺炎(AIT)大鼠甲状腺辅助性T细胞(Th)相关细胞因子γ干扰素(INF-γ)、白细胞介素10(IL-10)、白细胞介素17(IL-17)和白细胞介素35(IL-35)的调节作用,和对c-Jun氨基末端激酶(JNK)/促分裂素原活化蛋白激酶(p38 MAPK)信号通路的影响。方法40只SPF级雌性SD大鼠按随机数表分为正常组、模型对照组、夏枯草组和硒酵母组,每组均10只。除正常组外其他组使用高碘水(NaI)喂养联合注射猪甲状腺球蛋白(pTg)与弗氏佐剂(CFA、IFA)诱导AIT大鼠模型,造模后连续灌胃给药4周。实验过程中动态观察大鼠状态,包括一般情况和行为学情况,实验结束后取大鼠血清、甲状腺组织,ELISA检测大鼠甲状腺功能(FT3、FT4、TSH),甲状腺抗体(TPOAb、TGAb)及Th相关细胞因子(INF-γ、IL-10、IL-17、IL-35),HE染色观察大鼠甲状腺病理变化,Western blot检测大鼠甲状腺组织中磷酸化的JNK(P-JNK)、磷酸化的p38 MAPK(P-p38 MAPK)蛋白表达。结果与模型对照组比较,夏枯草及硒酵母组大鼠在体重、摄食量和粪便含水量方面均明显改善(P<0.05),在5min内运动距离、中央穿格次数和中央停留时间方面均明显下降(P<0.05),在淋巴细胞浸润和滤泡破坏上均有明显恢复。与模型对照组比较,夏枯草及硒酵母组大鼠血清FT3、FT4、TPOAb、TGAb、IFN-γ、IL-17水平降低(P<0.05),而TSH、IL-10、IL-35水平升高(P<0.05),甲状腺组织中p-JNK、p-p38 MAPK蛋白表达降低(P<0.05)。结论夏枯草对AIT大鼠甲状腺破坏具有保护作用,该作用与平衡Th相关细胞因子表达,抑制JNK/p38 MAPK信号通路的活化有关。

Objective To study the effects of Prunella vulgaris on Th cell-related cytokines INF-γ,IL-10,IL-17 and IL-35 and JNK/p38 MAPK signaling pathway in rats with autoimmune thyroiditis(AIT).Methods 40 SPF female SD rats were divided into normal group,model control group,Prunella vulgaris group,and selenium yeast group by random number table,with 10 rats in each group.The AIT rat models established by high-iodine water(NaI)feeding combined with injection of porcine thyroglobulin(pTg)and Freund's adjuvant were then intragastrically given the corresponding drugs for 4 weeks.During the experiment,the status of rats was dynamically observed,including general and behavioral conditions.After the experiment,the serum and thyroid tissues of rats were taken,the rats were subjected to the detection of serum levels of FT3,FT4,TSH,TPOAb,TGAb,INF-γ,IL-10,IL-17 and IL-35 by ELISA.The observation of the histopathological changes of thyroid tissues under light microscope by HE staining.The expressions of P-JNK and P-p38 MAPK proteins in thyroid tissues of rats were detected by WB.Results Compared with model control group,the body weight,food intake and fecal water content of Prunella vulgaris and selenium yeast groups were significantly improved(P<0.05),the movement distance within 5min,the number of central penetration and central residence time were significantly decreased(P<0.05),and the lymphocyte infiltration and follicular damage were significantly recovered.Compared with model control group,the groups intervened with Prunella vulgaris and selenium yeast decreased serum levels of FT3,FT4,TPOAb,TGAb,IFN-γand IL-17(P<0.05),increased TSH,IL-10 and IL-35(P<0.05),and decreased the protein expression of P-JNK and P-p38 MAPK in thyroid tissues(P<0.05).Conclusion Prunella vulgaris has a protective effect on thyroid damage in AIT rats,which is related to balance the expression of Th cell-related cytokine,and inhibit the excitation of JNK/p38 MAPK pathway.