穿山龙颗粒制剂对自身免疫性甲状腺炎大鼠TLR4/MyD88/NF-κB信号通路的影响  

Effect of Chuanshanlong Granule on TLR4/MyD88/NF-κB Signaling Pathway in Rats with Autoimmune Thyroiditis

作　　者：钟丰鹰 张程斐 秦慧钊 马知遥 吴丽丽[3,4] 秦灵灵[5] 刘铜华[3,4] ZHONG Fengying;ZHANG Chengfei;QIN Huizhao;MA Zhiyao;WU Lili;QIN Lingling;LIU Tonghua(Dongfang Hospital Beijing University of Chinese Medicine,Beijing 100078,China;School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488,China;Key Laboratory for Chinese Medicine Health Preservation of Ministry of Education,Beijing University of Chinese Medicine,Beijing 100029,China;Institute of Chinese Medicine Health Preservation,Beijing University of Chinese Medicine,Beijing 100029,China;Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学东方医院,北京100078 [2]北京中医药大学生命科学学院,北京102488 [3]北京中医药大学教育部中医养生学重点实验室,北京100029 [4]北京中医药大学中医养生学研究所,北京100029 [5]北京中医药大学,北京100029

出　　处：《中国实验方剂学杂志》2022年第16期42-49,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家国际科技合作专项项目(2015DFA30910);北京中医药大学校级科研纵向发展基金项目(2020-ZXFZJJ-005)。

摘　　要：目的:通过观察穿山龙颗粒制剂对Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核转录因子-κB(NF-κB)信号通路的影响,探讨其治疗自身免疫性甲状腺炎(AIT)大鼠的作用机制。方法:40只Lewis大鼠采用高碘饮水诱导和猪甲状腺球蛋白佐剂注射的方法制备AIT模型。6周后随机分为模型组、穿山龙低、中、高剂量组(0.52、1.03、2.06 g·kg^(-1)·d^(-1)),每组10只,分别予以0.01 mL·g^(-1)·d^(-1)相应体积混悬液,正常组及模型组大鼠予以同体积去离子水,灌胃8周后取材。取大鼠血清用酶联免疫吸附测定法(ELISA)检测甲状腺球蛋白抗体(TGAb)、甲状腺过氧化物酶抗体(TPOAb);检测游离三碘甲状腺原氨酸(FT3)、游离甲状腺素(FT4)、促甲状腺激素(TSH)的浓度;取大鼠甲状腺组织行苏木素-伊红(HE)染色,光镜下观察甲状腺组织的病理变化,并用免疫组化法及实时荧光定量聚合酶链式反应(Real-time PCR)测定TLR4、MyD88、NF-κB蛋白及mRNA的相对表达量。结果:与正常组比较,模型组大鼠血清中TPOAb、TGAb浓度均显著升高(P<0.01),FT3、FT4显著升高(P<0.01),TSH显著降低(P<0.01);与模型组比较,3个穿山龙剂量组的TPOAb、TGAb浓度均显著下降(P<0.01),穿山龙高剂量组FT3、FT4浓度显著下降(P<0.01),TSH显著升高(P<0.01)。HE染色示模型组甲状腺滤泡间隙存在淋巴细胞浸润,大量滤泡腔被破坏或变小,腔内胶质含量减少,滤泡壁变薄或被破坏,而穿山龙治疗组大鼠甲状腺组织内淋巴细胞浸润较模型组明显减少,甲状腺滤泡结构更为完整。与正常组比较,模型组的TLR4、MyD88、NF-κB蛋白表达显著上调(P<0.01),TLR4、MyD88 mRNA相对表达量显著上调(P<0.01),NF-κB mRNA相对表达量显著上调(P<0.01);与模型组比较,穿山龙高剂量组的TLR4蛋白及mRNA相对表达量显著下调(P<0.05),MyD88蛋白表达显著下调(P<0.01)、mRNA相对表达量显著下调(P<0.05),NF-κB蛋白及mRNA相对表达量均显著下调(P<Objective:To investigate the effect of Chuanshanlong granule on Toll-like receptor 4(TLR4)/myeloid differentiation protein 88(MyD88)/nuclear transcription factor-κB(NF-κB)signaling pathway,and to explore the mechanism of its treatment of autoimmune thyroiditis(AIT)in rats.Method:Forty AIT models were established following excess iodine and injection of porcine thyroglobulin and Freund's adjuvant into Lewis rats for six weeks.Then the rats were randomly divided into the model group,Chuanshanlong granule low-,medium-and high-dose group(0.52,1.03,2.06 g·kg^(-1)·d^(-1)),with ten in each group.Rats in the Chuanshanlong granule low-,medium-and high-dose groups were separately given 0.01 mL·g^(-1)·d^(-1) Chuanshanlong granule,and those in the normal group and the model group were given the same volume of deionized water for eight weeks.Serum of rats was taken to measure thyroglobulin antibody(TgAb)and thyroid peroxidase antibody(TPOAb)by enzyme-linked immunosorbent assay(ELISA),and the concentrations of free triiodothyronine(FT_(3)),free thyroxine(FT_(4))and thyroid stimulating hormone(TSH)were detected.The rat thyroid lobes were stained with hematoxylin and eosin(HE),and the pathological changes were observed under light microscope.In addition,the relative expression of TLR4,MyD88,NF-κB protein and mRNA was determined by immunohistochemistry and real-time polymerase chain reaction(Realtime PCR).Result:Compared with the conditions in the normal group,the serum concentrations of TPOAb and TgAb(P<0.01)and FT_(3) and FT_(4)(P<0.01)increased and TSH decreased(P<0.01)in the model group.Compared with the conditions in the model group,the concentrations of TPOAb and TgAb in the Chuanshanlong granule treatment groups reduced(P<0.01),and the concentrations of FT_(3) and FT_(4) were lowered(P<0.01)while TSH increased(P<0.01)in the Chuanshanlong granule high-dose group.HE staining showed that there was lymphocyte infiltration in the thyroid follicular space,a large number of destroyed or diminished follicular cavities,decre

关 键 词：穿山龙颗粒制剂 自身免疫性甲状腺炎 大鼠 Toll样受体4(TLR4)/髓样分化蛋白88(MyD88)/核转录因子-κB(NF-κB)信号通路 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33