地黄环烯醚萜苷类通过调控TGF-β1/Smads信号通路保护糖尿病小鼠肾脏作用  

Rehmanniae Radix Iridoid Glycosides Protect Kidneys of Diabetic Mice by Regulating TGF-β1/Smads Signaling Pathway

作　　者：张宏伟 刘明[2] 王慧森[2] 葛文静 张雪侠[2] 周倩[2] 李华妮 唐素勤[2] 李更生 ZHANG Hongwei;LIU Ming;WANG Huisen;GE Wenjing;ZHANG Xuexia;ZHOU Qian;LI Huani;TANG Suqin;LI Gengsheng(School of Pharmacy,Henan University,Kaifeng 475004,China;Institute of Traditional Chinese Medicine,Henan Integrative Medicine Hospital/Henan Academy of Chinese Medicine,Zhengzhou 450004,China;Academy of Chinese Medical Sciences,Henan University of Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南大学药学院,河南开封475004 [2]河南省中西医结合医院/河南省中医药研究院中药所,郑州450004 [3]河南中医药大学中医药科学院,郑州450046

出　　处：《中国实验方剂学杂志》2025年第2期56-66,共11页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省中医药科学研究专项重大课题(20-21ZYZD13);河南省中医药科学研究专项重点课题(2024ZY1026,2023ZY1023);河南省省属科研机构基本科研业务费项目(2304024,2204966)。

摘　　要：目的:观察地黄环烯醚萜苷类(RIG)成分对链脲佐菌素(STZ)诱导糖尿病小鼠肾脏的保护作用,并探讨其机制。方法:雄性C57BL/6J小鼠72只,随机抽取12只为正常组,其余60只进行高脂饲料喂养6周联合60 mg·kg^(-1)链脲佐菌素连续注射4 d进行2型糖尿病造模,造模成功后随机分为模型组、二甲双胍组(250 mg·kg^(-1))、梓醇组(100 mg·kg^(-1))、地黄环烯醚萜苷类低剂量组(RIG-L,200 mg·kg^(-1))和地黄环烯醚萜苷类高剂量组(RIG-H,400 mg·kg^(-1)),每组11只。各给药组小鼠灌胃对应药物,正常组和模型组给予等体积去离子水灌胃,1次/d。干预8周后,进行口服糖耐量(OGTT)实验,并计算OGTT曲线下面积(AUC),处死小鼠并剥离双肾组织。测定小鼠体质量、双肾质量、空腹血糖(FBG);生化试剂盒检测血清甘油三酯(TG)、胆固醇(TC)、肌酐(SCr)、尿素氮(BUN)含量;酶联免疫吸附测定法(ELISA)检测血清胰岛素(FINS)含量,计算胰岛素敏感指数(ISI)和胰岛素抵抗指数(HOMA-IR);苏木素-伊红(HE)染色、马松(Masson)染色观察肾脏组织病理变化;免疫组化法(IHC)检测肾脏组织白细胞介素-1(IL-1)、白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α)、转化生长因子-β(1 TGF-β1)、胶原蛋白-3(ColⅢ)的表达情况;蛋白免疫印迹法(Western blot)检测肾组织TGF-β1、细胞信号转导分子3(Smad3)、基质金属蛋白酶-9(MMP-9)、ColⅢ蛋白表达。结果:与正常组比较,模型组小鼠体质量、ISI显著降低(P<0.01),肾脏绝对质量、FBG、AUC、FINS、HOMA-IR、TC、TG、SCr、BUN水平显著升高(P<0.01),肾组织出现肾小球肥大、肾小囊腔窄小及胶原沉积,肾组织IL-1、IL-6、TNF-α、TGF-β1、ColⅢ、Smad3蛋白表达水平显著升高(P<0.01),MMP-9蛋白表达水平显著降低(P<0.01);与模型组比较,各给药组小鼠体质量变化差异无统计学意义;各给药组小鼠肾脏绝对质明显降低(P<0.05,P<0.01);RIG-H组小鼠给药第4~8周及二甲双胍、�Objective:To investigate the protective effect of Rehmanniae Radix iridoid glycosides(RIG)on the kidney tissue of streptozotocin(STZ)-induced diabetic mice and explore the underlying mechanism.Methods:Twelve of 72 male C57BL/6J mice were randomly selected as the normal group,and the remaining 60 mice were fed with a high-fat diet for six weeks combined with injection of 60 mg·kg^(-1)STZ for 4 days to model type 2 diabetes mellitus.The successfully modeled mice were randomized into model,metformin(250 mg·kg^(-1)),catalpol(100 mg·kg^(-1)),low-dose RIG(RIG-L,200 mg·kg^(-1))and high-dose RIG(RIG-H,400 mg·kg^(-1))groups(n=11).Mice in each group were administrated with corresponding drugs,while those in the normal group and model group were administrated with the same dose of distilled water by gavage once a day.After 8 weeks of intervention,an oral glucose tolerance test(OGTT)was performed,and the area under the curve(AUC)was calculated.After mice were sacrificed,both kidneys were collected.The body weight,kidney weight,and fasting blood glucose(FBG)were measured.Biochemical assays were performed to measure the serum levels of triglycerides(TG),total cholesterol(TC),serum creatinine(SCr),and blood urea nitrogen(BUN).Enzyme-linked immunosorbent assay(ELISA)was employed to determine the serum level of fasting insulin(FINS),and the insulin sensitivity index(ISI)and homeostatic model assessment for insulin resistance(HOMA-IR)were calculated.The pathological changes in kidneys of mice were observed by hematoxylin-eosin staining and Masson staining.The immunohistochemical method(IHC)was employed to assess the expression of interleukin-1(IL-1),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),transforming growth factor-β1(TGF-β1),and collagen-3(ColⅢ)in the kidney tissue.The protein levels of TGF-β1,cell signal transduction molecule 3(Smad3),matrix metalloproteinase-9(MMP-9),and ColⅢ in kidneys of mice were determined by Western blot.Results:Compared with the normal group,the model group showcased decreased bod

关 键 词：地黄环烯醚萜苷类 2型糖尿病 转化生长因子-β(1 TGF-β1)/细胞信号转导分子3(Smad3) 胶原蛋白-3(ColⅢ) 基质金属蛋白酶-9(MMP-9) 

分 类 号：R285[医药卫生—中药学] R243[医药卫生—中医学] R259