Effects of herbal cake-partitioned moxibustion on the expression of thyroid autophagy-related factors LC3B and Beclin-1 in rats with autoimmune thyroiditis  

隔药饼灸对自身免疫性甲状腺炎大鼠甲状腺自噬相关因子LC3B及Beclin-1表达的影响

作　　者：CHEN Kexu ZHAO Jimeng QIAO Yu ZHU Lu HUANG Yan LIU Yanan ZHENG Handan LIU Huirong CUI Yunhua WU Huangan YANG Yanping 陈珂旭;赵继梦;乔宇;朱璐;黄艳;刘雅楠;郑寒丹;刘慧荣;崔云华;吴焕淦;杨燕萍(Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China;Shanghai Research Institute of Acupuncture and Meridian,Shanghai 200030,China;不详)

机构地区：[1]Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China [2]Shanghai Research Institute of Acupuncture and Meridian,Shanghai 200030,China [3]不详

出　　处：《Journal of Acupuncture and Tuina Science》2022年第6期453-463,共11页针灸推拿医学（英文版）

基　　金：This work was supported by the National TCM Leading Talents Support Program-Qihuang Scholars(国家中医药领军人才支持计划-岐黄学者);National Program on Key Basic Research Project“973 Program”(国家重点基础研究发展计划“973计划项目”,No.2015CB554501);Projects of National Natural Science Foundation of China(国家自然科学基金项目,No.82074551,No.81704176);Youth Project from Department of Science and Education,Shanghai Health Commission(上海市卫健委科教处青年基金,No.20194Y0013).

摘　　要：Objective:To observe the anti-inflammatory effect,as well as the effect on the expression of microtubule-associated protein light chain 3B(LC3B)and Beclin-1 of herbal cake-partitioned moxibustion in rats with experimental autoimmune thyroiditis(EAT).Methods:Female Sprague-Dawley rats were randomly divided into a normal group and a modeling group.The EAT rat model was prepared by a combination of antigen immunization plus iodine agent induction.After the model was prepared,rats in the modeling group were randomly and equally divided into a model group and a herbal cake-partitioned moxibustion group.In the herbal cake-partitioned moxibustion group,moxibustion was alternately applied to two groups of points[Dazhui(GV14)-Mingmen(GV4)and Tiantu(CV22)-Guanyuan(CV4)],and the treatment continued for 30 d.Rats in the normal and model groups were only fixed identically without intervention.Histopathological manifestations of thyroid glands were observed by hematoxylin-eosin staining;the concentrations of thyroid peroxidase antibodies(TPOAb),thyroglobulin antibodies(TGAb),interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay;real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry were used to detect the mRNA and protein expression of autophagy-related factors LC3B and Beclin-1 in thyroid tissue.Results:There were massive follicular destruction,lymphocytic infiltration,and interstitial fibrous tissue hyperplasia of the thyroid glands in the model group.Some follicles of the thyroid glands were destroyed with few lymphocyte infiltrations and fibrous tissue hyperplasia in the moxibustion group.Compared with the normal group,the concentrations of serum TPOAb,TGAb,IL-1β,IL-6,and TNF-αwere increased in the model rats(P<0.05);the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were reduced in the model group(P<0.05).Compared with the model group,the concentrations of serum TPOAb,TGAb,IL-1β,IL-6,a目的:观察隔药饼灸干预实验性自身免疫性甲状腺炎(EAT)大鼠的抗炎效应,以及对自噬相关因子微管相关蛋白轻链3B(LC3B)及Beclin-1表达的影响.方法:将雌性Sprague-Dawley大鼠随机分为正常组和造模组,采用抗原免疫加碘剂联合诱导的方法制备EAT大鼠模型.模型制备完成后将造模组大鼠随机等分为模型组和隔药饼灸组.隔药饼灸组在两组穴位(大椎-命门和天突-关元)交替进行隔药饼灸治疗,连续干预30 d.正常组和模型组只做相同固定,不进行干预治疗.采用苏木素-伊红染色法观察甲状腺组织病理学表现;采用酶联免疫吸附剂测定法检测血清抗甲状腺过氧化物酶抗体(TPOAb)、抗甲状腺球蛋白抗体(TGAb)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的浓度;采用实时荧光定量多聚核苷酸链式反应法和免疫组化法检测甲状腺组织自噬相关因子LC3B和Beclin-1的mRNA及蛋白表达.结果:模型组甲状腺大量滤泡破坏,有大量淋巴细胞浸润,间质大量纤维组织增生;隔药饼灸组甲状腺部分滤泡破坏,有少量淋巴细胞浸润,少量纤维组织增生.与正常组比较,模型组大鼠血清TPOAb、TGAb、IL-1β、IL-6和TNF-α的浓度均升高(P<0.05);模型组甲状腺组织LC3B和Beclin-1的mRNA与蛋白表达水平均降低(P<0.05).与模型组相比,隔药饼灸组大鼠血清TPOAb、TGAb、IL-1β、IL-6和TNF-α的浓度均降低(P<0.05);隔药饼灸组甲状腺组织LC3B和Beclin-1的mRNA与蛋白表达水平均升高(P<0.05).甲状腺组织LC3B和Beclin-1的mRNA及蛋白表达水平与血清TPOAb及TGAb的浓度均呈负相关.结论:隔药饼灸能减轻EAT大鼠甲状腺的炎性反应,降低血清TPOAb及TGAb水平.这可能与调节大鼠甲状腺组织自噬相关因子LC3B和Beclin-1的mRNA及蛋白表达有关.

关 键 词：Moxibustion Therapy Medicinal Cake-partitioned Moxibustion Thyroiditis Autoimmune AUTOPHAGY Points Conception Vessel Points Governor Vessel RATS 

分 类 号：R2-03[医药卫生—中医学]