基于TGF-β1/Smad3信号通路探讨黄芪甲苷对甲状腺功能减退幼鼠肾损伤的改善作用  

Ameliorative Effects of Astragaloside-Ⅳon Renal Injury in Infant Rats with Hypothyroidism Based on TGF-β1/Smad3 Signaling Pathway

作　　者：张晖[1] 刘鹏[1] 常毅娜[1] 李晓乐[2] ZHANG Hui;LIU Peng;CHANG Yina;LI Xiaole(The First Affiliated Hospital of Henan University of Science and Technology,Luoyang 471000,China;The Third Affiliated Hospital of Henan University,Zhengzhou 450000,China)

机构地区：[1]河南科技大学第一附属医院,河南洛阳471000 [2]郑州大学第三附属医院,河南郑州450000

出　　处：《中医药信息》2022年第7期11-17,共7页Information on Traditional Chinese Medicine

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20190357)。

摘　　要：目的:探讨黄芪甲苷(AS-Ⅳ)通过介导转化生长因子-β1(TGF-β1)/Smad3信号通路对甲状腺功能减退(HT)幼鼠肾损伤的作用。方法:随机选取10只30日龄雄性Wistar大鼠作为正常组,剩余50只幼鼠连续28 d灌胃6 mg/kg剂量的6-丙基-2-硫尿嘧啶(PTU)以诱导HT模型。成功建模43只,随机剔除3只后剩余大鼠分为模型组、AS-Ⅳ低剂量组、AS-Ⅳ高剂量组及左甲状腺素钠(LS)组,每组各10只。建模成功次日给药,其中AS-Ⅳ低、高剂量组及LS组幼鼠灌胃给药量分别为20、40 mg/(kg·d)和5μg/(kg·d),正常组及模型组大鼠灌胃等体积生理盐水,每日1次,连续5周。给药结束后采用Bradford法检测24 h尿蛋白(Alb)含量;测定肾脏脏器指数;HE染色法观察肾组织形态学变化;Masson染色法观察肾组织胶原蛋白沉积情况;ELISA法检测甲状腺功能指标即血清游离三碘甲腺原氨酸(FT3)、游离甲状腺素(FT4)及促甲状腺素(TSH)水平;生化分析仪检测肾功能指标即血清肌酐(Scr)及尿素氮(BUN)含量;Western blot法检测肾组织中转化生长因子-β1(TGF-β1)、TGF-β受体I型(TβRI)、Smad3、p-Smad3、Smad7、α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白(ColⅠ)蛋白相对表达情况。结果:本研究成功构建HT幼鼠模型;与正常组比较,模型组幼鼠血清FT3、FT4水平降低,TSH水平升高,肾脏脏器系数、24 h尿Alb含量、Scr及BUN含量升高(P<0.05),肾组织中可见明显肾间质损伤,有大量胶原纤维表达和沉积,肾组织中TGF-β1、TβRI、α-SMA、ColⅠ蛋白相对表达水平及p-Smad3/Smad3升高,Smad7蛋白相对表达水平降低(P<0.05);与模型组比较,AS-Ⅳ低、高剂量组及LS组幼鼠血清FT3、FT4水平升高,TSH水平降低,肾脏脏器系数降低、24 h尿Alb含量Scr及BUN含量降低(P<0.05),肾组织结构病理损伤逐渐改善,胶原纤维沉积情况逐渐减轻,肾组织中TGF-β1、TβRI、α-SMA、ColⅠ蛋白相对表达水平及p-Smad3/Smad3降低,Smad7蛋白相对表达水�Objective:To investigate the effect of astragalosideⅣ(AS-Ⅳ)improving renal injury in infant rats with hypothyroidism(HT)through mediating transforming growth factor-β1(TGF-β1)/Smad3 signaling pathway.Methods:10 male Wistar rats of 30-day-old were randomly selected as the normal control,and the remaining 50 rats were given 6-propyl-2-thiouracil(PTU,6 mg/kg)intragastric administration for 28 days to induce HT model.40 successful modeling rats were randomly divided into the model group,the low-dose AS-Ⅳgroup[20 mg(kg·d)],the high-dose AS-Ⅳgroup[40 mg(kg·d)]and the Levothyroxine sodium group[LS,5μg(kg·d)],with 10 rats in each group.After five weeks of continuous treatment,Bradford method was used to detect 24 h urinary protein(Alb),renal organ coefficient was determined,HE staining was used to observe the morphological changes of renal tissues,Masson staining was used to observe the collagen deposition in renal tissues,the serum levels of free triiodothyronine(FT3),free thyroxine(FT4)and thyrotropin stimulating hormone(TSH)were detected by ELISA,the serum contents of creatinine(Scr)and urea nitrogen(BUN)were detected by biochemical analyzer,and the relative protein expressions of transforming growth factor-β1(TGF-β1),TGF-βreceptor type I(TβRI),Smad3,p-SMad3,Smad7,α-smooth muscle actin(α-SMA)and collagen type I(Col I)in renal tissues were detected by Western blot method.Results:The HT infant rat model was successfully constructed in this study.Compared with those in the normal control group,the serum levels of FT3 and FT4 were decreased,the serum level of TSH was increased,the renal organ coefficient was increased,and the contents of 24 h urine Alb,Scr and BUN were increased in the model group(P<0.05);obvious renal interstitial injury,abundant collagen fiber expression and deposition were observed in the model group;the relative protein expressions of TGF-β1,TβRI,α-SMA,Col I,Smad3 and p-Smad3 were increased,whereas the relative protein expression of Smad7 was decreased in the model group(P<0.05)

关 键 词：黄芪甲苷 转化生长因子-Β SMAD 甲状腺功能减退 肾损伤 

分 类 号：R285.5[医药卫生—中药学]