基于Akt/mTOR通路观察补中益气颗粒对实验性自身免疫性甲状腺炎大鼠的保护作用  

Protective effect of Buzhong Yiqi granule on experimental autoimmune thyroiditis in rats and its effect on Akt/mTOR pathway

作　　者：姚婷 万明 YAO Ting;WAN Ming(The Second Affiliated Hospital of Chengdu Medical College,Chengdu 610051,China)

机构地区：[1]成都医学院第二附属医院核工业四一六医院,四川成都610051

出　　处：《陕西中医》2023年第2期159-164,共6页Shaanxi Journal of Traditional Chinese Medicine

基　　金：四川省医学科研计划课题(S20083)。

摘　　要：目的:基于蛋白激酶B(Akt)/雷帕霉素靶向基因(mTOR)信号通路研究补中益气颗粒对实验性自身免疫性甲状腺炎(EAT)大鼠的保护作用机制。方法:取30只SPF级SD雌性大鼠采用猪甲状腺球蛋白(pTg)和完全弗氏佐剂(CFA)混合免疫注射联合高碘喂养法制备EAT大鼠模型,并随机分为模型组、低剂量组(低剂量补中益气颗粒组)和高剂量组(高剂量补中益气颗粒组),另取10只大鼠相同部位注射等体积PBS溶液及普通动物饮用水喂养作为空白组。低剂量组和高剂量组分别按0.80、0.94g/kg的剂量灌胃补中益气汤,其余两组给予等体积的双蒸水,造模结束后,以血清甲状腺过氧化物酶抗体(TPOAb)、甲状腺球蛋白抗体(TGAb)和促甲状腺激素(TSH)水平进行模型评价。药物治疗结束当天,酶联免疫吸附检测(ELISA)法检测大鼠血清甲状腺功能指标和抗体水平;观察甲状腺组织病理形态和滤泡上皮细胞的自噬情况;免疫组化观察甲状腺组织中γ干扰素(IFN-γ)、白细胞介素-4(IL-4)、半胱氨酸蛋白酶3(Caspase-3)表达水平;Western blot检测Akt/mTOR通路蛋白表达。结果:造模组大鼠造模完成后的血清TPOAb、TGAb、TSH水平明显高于空白组(P<0.05),提示造模成功。模型组大鼠治疗后的血清TPOAb、TGAb、TSH、FT3、FT4、T3、T4水平、甲状腺组织IFN-γ、Caspase-3、p-Akt、p-mTOR蛋白表达水平、IFN-γ/IL-4值较空白组明显增加,甲状腺滤泡上皮细胞自噬小体数量明显减少(P<0.05);与模型组比较,低剂量组和高剂量组大鼠血清TPOAb、TGAb、TSH、FT3、FT4、T3、T4水平、甲状腺组织IFN-γ、Caspase-3、p-Akt、p-mTOR蛋白表达水平、IFN-γ/IL-4值明显下降,甲状腺滤泡上皮细胞自噬小体数量明显增加(P<0.05);四组大鼠甲状腺组织中IL-4、Akt、mTOR表达水平差异无统计学意义(P>0.05)。结论:补中益气颗粒对EAT大鼠起到保护作用,可改善其甲状腺功能和自身抗体水平,恢复甲状腺组织�Objective:Protective mechanism of Buzhong Yiqi granule on EAT rats was studied based on Akt/mTOR signaling pathway.Methods:A total of 30 female SPF SD rats were injected with porcine thyroglobulin(pTg)and CFA combined with high iodine feeding method to establish EAT rat model,and randomly divided into model group,low-dose group(low-dose Buzhong Yiqi granule group)and high-dose group(high-dose Buzhong Yiqi granule group).Another 10 rats were injected with the same volume of PBS solution and common animal drinking water as blank group.Low and high-dose groups were given Buzhong Yiqi decoction at the dose of 0.80 and 0.94g/kg,respectively,and the other two groups were given an equal volume of double distilled water.Success of EAT rat model was evaluated by the levels of serum thyroid peroxidase antibody(TPOAb),thyroglobulin antibody(TGAb)and thyroid stimulating hormone(TSH).On the day after drug treatment,serum thyroid function index and antibody level were detected by ELISA,pathological morphology of thyroid tissue was detected by hematoxylin-eosin(HE)staining,and autophagy of thyroid follicular epithelial cells was observed by transmission electron microscope.Eexpression of interferon-gamma(IFN-γ),interleukin-4(IL-4)and cysteine protease-3(Caspase-3)in thyroid tissue was detected by immunohistochemistry,and expression of Akt/mTOR pathway protein was detected by Western blot.Results:After the completion of the model,compared with the blank group,serum TPOAb,TGAb and TSH levels of the rats in the modeling group were significantly increased(P<0.05).After treatment,levels of serum TPOAb,TGAb,TSH,free triiodothyronine(FT3),FT4,triiodothyronine(T3),thyroxine(T4),IFN-γ,Caspase-3,p-Akt,p-mTOR protein expression and IFN-γ/IL-4 ratio in the model group were significantly higher than those in the blank group,while the number of autophagy bodies in thyroid follicular epithelial cells decreased significantly(P<0.05).Levels of serum TPOAb,TGAb,TSH,FT3,FT4,T3,T4,IFN-γ,Caspase-3,p-Akt,p-mTOR protein expression and IFN-γ/IL-4

关 键 词：自身免疫性甲状腺炎 蛋白激酶B/雷帕霉素靶向基因信号通路 补中益气颗粒 自噬小体 大鼠 甲状腺功能 

分 类 号：R581.4[医药卫生—内分泌]