摘要
目的:探讨补脾强力复方对实验性重症肌无力伴亚临床甲减大鼠的治疗作用及对下丘脑-垂体-甲状腺-胸腺(HPTT)轴的影响。方法:选择SPF级Lewis大鼠,随机分为正常组、模型组、补脾强力复方低剂量、中剂量及高剂量组。模型大鼠通过乙酰胆碱受体α亚基97-116肽段免疫接种构建实验性重症肌无力模型后再通过甲巯咪唑制备实验性重症肌无力伴亚临床甲减模型。补脾强力复方低、中、高剂量组分别以4.57 g/kg、7.12 g/kg、9.49 g/kg补脾强力复方灌胃,正常组和模型组则给予等体积生理盐水灌胃,每日1次,连续灌胃4周。灌胃结束后,记录各组大鼠Lennon评分;检测HPTT轴相关病理及分子水平变化,其中通过HE染色检测胸腺及甲状腺组织病理变化,ELISA检测血清中乙酰胆碱受体抗体(AchRab)、促甲状腺激素(TSH)、促甲状腺素释放激素(TRH)、游离甲状腺素(FT4)及甲状腺素(T4)表达水平,qPCR检测下丘脑中TRH及垂体组织中TSH的mRNA水平,Western blot检测胸腺中Cleaved Caspase3、Bcl2相关X蛋白(Bax)及B细胞淋巴瘤基因2(Bcl2)蛋白表达。结果:与正常组大鼠相比,模型组大鼠肌无力症状明显,Lennon评分显著升高(P<0.05),胸腺及甲状腺组织出现明显病理改变,血清中FT4及T4表达无明显变化(P>0.05),血清中AchRab、TSH及TRH表达显著增加(P<0.05),下丘脑中TRH及垂体中TSH表达显著增加(P<0.05),胸腺中Cleaved Caspase3及Bax蛋白表达显著减少(P<0.05),胸腺中Bcl2蛋白表达显著增加(P<0.05);与模型组大鼠相比,补脾强力复方高剂量组大鼠肌无力症状有所改善,Lennon评分显著降低(P<0.05),胸腺及甲状腺组织病理变化有所改善,血清中FT4及T4表达无明显变化(P>0.05),血清中AchRab、TSH及TRH表达显著减少(P<0.05),下丘脑中TRH及垂体中TSH表达显著减少(P<0.05)、胸腺中Cleaved Caspase3及Bax表达显著增加(P<0.05)、胸腺中Bcl2表达显著减少(P<0.05)。结论:补脾强力复方可改善实验性重症肌无力伴亚临床甲减大鼠的临床症状,其作用机制可能与调节HPTT轴有关。
Objective:To explore the therapeutic effect of Bupiqiangli compound on experimental myasthenia gravis with sub-clinical hypothyroidism in rats and the influence of hypothalamus-pituitary-thyroid-thymus(HPTT)axis.Methods:SPF Lewis rats were randomly divided into normal group,model group,Bupiqiangli compound low-dose,medium-dose and high-dose groups.Model rats were immunized with AchR-αsubunit 97-116 peptide sequence to construct an experimental myasthenia gravis model,and then methimazole was used to prepare an experimental myasthenia gravis with subclinical hypothyroidism model.Bupiqiangli compound low-dose,medium-dose and high-dose groups were given 4.57 g/kg,7.12 g/kg and 9.49 g/kg of Bupiqiangli compound by gavage,nor-mal group and model group were given an equal volume of normal saline by gavage,once a day,for 4 weeks.After the last gavage,Lennon scores of rats in each group were recorded;HE staining was used to detect pathological changes of thymus and thyroid;ELISA was used to detect expression levels of acetylcholine receptor antibody(AchRab),thyroid stimulating hormone(TSH),thyrotropin releasing hormone(TRH),free thyroxine(FT4)and thyroxine(T4)in serum;mRNA level of TRH in hypothalamus and TSH in pituitary tissue were detected by qPCR;Western blot detected changes of protein expressions of Cleaved Caspase3,Bcl2 associated X protein(Bax)and B-cell lymphoma-2(Bcl2)in thymus.Results:Compared with normal group,model group showed obvious symp-toms of muscle weakness,Lennon score increased significantly(P<0.05),obvious pathological changes in thymus and thyroid tissues,while no significant changes in expressions of FT4 and T4 in serum(P>0.05),expressions of AchRab,TSH and TRH in serum were significantly increased(P<0.05),expressions of TRH in hypothalamus and TSH in pituitary were significantly increased(P<0.05),protein expressions of Cleaved Caspase3 and Bax in thymus were significantly decreased(P<0.05),while expression of Bcl2 protein in thymus increased significantly(P<0.05).Compared with model group,myasthenia symptoms of compound high-dose group were im-proved,Lennon score was significantly decreased(P<0.05),pathological changes of thymus and thyroid tissues were improved,ex-pressions of FT4 and T4 in serum had no significant changes(P>0.05),expressions of AchRab,TSH and TRH in serum were signifi-cantly decreased(P<0.05),expressions of TRH in hypothalamus and TSH in pituitary were significantly decreased(P<0.05),expres-sions of Cleaved Caspase3 and Bax in thymus were significantly increased(P<0.05),while expression of Bcl2 in thymus was signifi-cantly decreased(P<0.05).Conclusion:Bupiqiang compound can improve clinical symptoms of experimental myasthenia gravis with subclinical hypothyroidism in rats,and its mechanism may be related to the regulation of HPTT axis.
作者
王强
李若照
况时祥
赵海
钱忆家
雍波
郭菁
刘运权
WANG Qiang;LI Ruozhao;KUANG Shixiang;ZHAO Hai;QIAN Yijia;YONG Bo;GUO Jing;LIU Yunquan(The Second Clinical Medical College of Guizhou University of Traditional Chinese Medicine,Guiyang 550000,China)
出处
《中国免疫学杂志》
北大核心
2025年第8期1806-1811,共6页
Chinese Journal of Immunology
基金
国家自然科学基金项目(81960856)
贵州省科学技术基金(黔科合平台人才[2018]5605号,黔科合J字[2015]2022号,黔教合KY字[2017]178号)
贵州省中医药管理局项目(QZYY-2021-126)。
