基于甲状腺激素合成信号通路CGA/GPX2/TSHβ探讨逍遥散对肝郁脾虚证大鼠HPT轴的影响  

Xiaoyaosan Regulates HPT Axis in Rat Model with Syndrome of Liver Depression and Spleen Deficiency via CGA/GPX2/TSHβPathway for Thyroid Hormone Synthesis

作　　者：王芳 袁汝鑫 樊玲瑾 陈宗礼 肖华业[1] 杨力强[1,2] 李晓红[1,2] 郑春成[1] WANG Fang;YUAN Ruxin;FAN Lingjin;CHEN Zongli;XIAO Huaye;YANG Liqiang;LI Xiaohong;ZHENG Chuncheng(Guangxi University of Chinese Medicine,Nanning 530200,China;Guangxi Key Laboratory of Chinese Medicine Foundation Research,Guangxi University of Chinese Medicine,Nanning 530200,China)

机构地区：[1]广西中医药大学,南宁530200 [2]广西中医药大学广西中医基础研究重点实验室,南宁530200

出　　处：《中国实验方剂学杂志》2025年第3期1-10,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82360900,81860817);广西自然科学基金项目(2018GXNSFAA138018);广西中医药大学广西中医基础研究重点实验室自主研究课题(20-065-53-06)。

摘　　要：目的:通过观察逍遥散对肝郁脾虚证大鼠甲状腺激素合成通路糖蛋白激素α亚基/谷胱甘肽过氧化酶2/促甲状腺素β亚基(CGA/GPX2/TSHβ)的影响,探讨其调节肝郁脾虚证大鼠HPT轴功能紊乱的作用机制。方法:72只雄性SD大鼠按随机数字表法分为正常组、模型组、逍遥散高、中、低剂量组(16.7、8.35、4.175 g·kg^(-1))、氟西汀组(0.0018 g·kg^(-1)),每组12只。用慢性束缚应激21 d的方法复制肝郁脾虚证大鼠模型,逍遥散各剂量组和氟西汀组分别用逍遥散中药煎剂和氟西汀悬浊液干预。造模结束后苏木素-伊红(HE)染色观察大鼠甲状腺、垂体组织形态学变化;酶联免疫吸附测定法(ELISA)测定血清三碘甲状腺原氨酸(T3)、四碘甲状腺原氨酸(T4)、促甲状腺激素(TSH)含量;实时荧光定量聚合酶链式反应(Real-time PCR)、蛋白免疫印迹法(Western blot)检测各组大鼠甲状腺组织促甲状腺激素受体(TSHR)、垂体组织促甲状腺激素释放激素受体(TRHR)和TSHβ及下丘脑组织促甲状腺激素释放激素(TRH)、CGA、GPX2、TSHβmRNA和蛋白表达。结果:与正常组比较,模型组大鼠甲状腺大量腺泡萎缩,不规则,胶体分泌严重减少,垂体前叶大量腺细胞空泡变性,血清T3、T4、TSH含量显著降低(P<0.01),甲状腺组织中TSHR、垂体组织中TRHR与TSHβ、下丘脑组织中TRH、CGA、GPX2、TSHβmRNA及蛋白表达显著降低(P<0.01);与模型组比较,逍遥散高、中剂量组、氟西汀组大鼠甲状腺与垂体组织病理变化得到明显改善优于逍遥散低剂量组;逍遥散高、中剂量组、氟西汀组血清T3、T4、TSH含量均明显升高(P<0.05,P<0.01),逍遥散低剂量组血清TSH含量亦明显升高(P<0.05);逍遥散高、中剂量组大鼠甲状腺TSHR、垂体TRHR、TSHβ、下丘脑TRH、CGA、GPX2、TSHβ的mRNA和蛋白表达水平均明显升高(P<0.05,P<0.01),逍遥散低剂量组大鼠下丘脑TSHβ的mRNA和蛋白表达显著升高(P<0.01),氟西汀组大�Objective:To explore the mechanism by which Xiaoyaosan regulates HPT axis dysfunction in the rat model with the syndrome of liver depression and spleen deficiency by observing its effect on the glycoprotein hormoneα-subunit(CGA)/glutathione peroxidase 2(GPX2)/thyroid-stimulating hormoneβ-subunit(TSHβ)pathway for thyroid hormone synthesis.Methods:Seventy-two male SD rats were randomized into six groups:normal,model,high-dose(16.7 g·kg^(-1)),medium-dose(8.35 g·kg^(-1)),and low-dose(4.175 g·kg^(-1))Xiaoyaosan,and fluoxetine(0.0018 g·kg^(-1))groups,with 12 rats in each group.The rat model of liver depression and spleen deficiency was induced by chronic restraint stress for 21 days.The intervention groups were treated with Xiaoyaosan decoctions or fluoxetine suspension,respectively.After modeling,hematoxylin-eosin staining was employed to observe morphological changes in the thyroid and pituitary tissue of the rats.Serum levels of triiodothyronine(T3),tetraiodothyronine(T4),and thyroid-stimulating hormone(TSH)were measured by enzyme-linked immunosorbent assay(ELISA).Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)and Western blot were employed to determine the mRNA and protein levels,respectively,of TSH receptor(TSHR)in the thyroid tissue,thyrotropin-releasing hormone receptor(TRHR)and TSHβin the pituitary tissue,and thyrotropin-releasing hormone(TRH),CGA,GPX2,and TSHβin the hypothalamic tissue.Results:Compared with the normal group,the model group showed significant atrophy and irregularity of thyroid follicles,a marked reduction in colloid secretion,extensive vacuolar degeneration of adenocytes in the anterior pituitary,lowered serum levels of T3,T4,and TSH(P<0.01),and down-regulated mRNA and protein levels of TSHR in the thyroid tissue,TRHR and TSHβin the pituitary tissue,and TRH,CGA,GPX2,and TSHβin the hypothalamic tissue(P<0.01).Compared with the model group,high-and medium-dose Xiaoyaosan and fluoxetine alleviated the pathological changes in the thyroid and pituitary tissue

关 键 词：慢性束缚应激 肝郁脾虚证 糖蛋白α亚基/谷胱甘肽过氧化物酶2/促甲状腺素β亚基(CGA/GPX2/TSHβ)信号通路 逍遥散 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学] R256