腹腔注射黄芪注射液的糖尿病小鼠肾脏组织病理变化观察  

Histopathological changes of kidney tissues in diabetic mice with intraperitoneal injection of astragalus injection

作　　者：黄云芳 付会玲 张云 刘昌璇[1] 贾琳 黄敏 陈文莉[1] Huang Yunang1, FU Huilin, ZHANG Yun, LIU Changxuan, JIA Lin, HUANG Min, CHEN Wenli(The Central Hospital of Wuhan, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430014, China)

机构地区：[1]华中科技大学同济医学院附属武汉中心医院,武汉430014 [2]武汉市第六人民医院

出　　处：《山东医药》2018年第31期30-34,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81400734);武汉市卫计委科研项目(WX13B06)

摘　　要：目的观察腹腔注射黄芪注射液的糖尿病小鼠肾脏组织病理变化,并探讨其可能作用机制。方法将C57小鼠随机分为四组。正常对照组小鼠不经任何干预。黄芪组、贝那普利组、糖尿病组制作糖尿病模型。造模成功后黄芪组腹腔注射黄芪注射液0.03 m L/10 g,1次/d;贝那普利组予贝那普利灌胃,10 mg/kg,1次/d,糖尿病组小鼠不做任何处理。饲养12周收集各组小鼠静脉血,取各组肾脏组织。PAS染色后镜下观察各组肾脏病理情况,ELISA法检测各组肾脏组织、血清血管紧张素Ⅱ(AngⅡ),免疫组化染色法检测肾脏组织α-平滑肌肌动蛋白(α-SMA)、Ⅳ型胶原(Collagen-Ⅳ),Masson染色法观察各组肾脏组织纤维化情况,免疫组化和Real-time PCR法检测各组肾脏组织血管紧张素转化酶(ACE)、ACE2蛋白及mRNA,硫代巴比妥酸法检测各组肾脏组织丙二醛(MDA)含量,Real-time PCR法检测各组肾脏组织MCP-1、ICAM-1、TGF-β和TNF-αmRNA。结果与正常对照组小鼠相比,糖尿病小鼠肾脏组织肾小球肥大、系膜基质增多和系膜细胞增生的病理改变,黄芪组小鼠肾脏组织和贝那普利组小鼠肾脏组织肾小球肥大、系膜基质增多和系膜细胞增生情况减轻。与正常对照组相比,糖尿病组肾脏组织AngⅡ、α-SMA、Collagen-Ⅳ相对表达量均升高(P<0.05),与糖尿病组相比,黄芪组、贝那普利组肾脏组织AngⅡ、α-SMA、Collagen-Ⅳ相对表达量均降低(P均<0.05)。与正常对照组比较,糖尿病组、黄芪组、贝那普利组ACE mRNA及蛋白相对表达量均降低,ACE2 mRNA及蛋白相对表达量均升高(P均<0.05);与糖尿病组比较,贝那普利组ACE mRNA及蛋白相对表达量均升高,ACE2 mRNA及蛋白相对表达量均降低(P均<0.05);黄芪组ACE蛋白相对表达量升高,ACE2 mRNA及蛋白相对表达量均降低(P均<0.05)。与正常对照组比较,糖尿病组、黄芪组、贝那普利组肾脏组织MDA含量均升高(P均<0.05);与糖尿病组比较,Objective To observe the pathological changes of kidney tissues in diabetic mice with intraperitoneal injection of astragalus,and to explore its possible mechanism. Methods C57 mice were randomly divided into four groups.The mice in the normal control group were not subjected to any intervention. We established the diabetic models in the astragalus group,benazepril group,and diabetic group. After the successful modeling,mice in the astragalus group received intraperitoneal injection of 0. 03 m L/10 g astragalus,once daily; mice in the benazepril group were administered 10 mg/kg benazepril,once daily,and mice in the diabetic group did not receive any treatment. The venous blood of each group was collected after 12 weeks,and the kidney tissues of each group were taken. After PAS staining,the pathological conditions of the kidneys were observed under the microscope. The renal tissues and serum angiotensin II（Ang II） were detected by ELISA. The α-smooth muscle actin（α-SMA） and Collagen IV were detected by immunohistochemical staining. Masson staining was used to observe the fibrosis of renal tissues in each group. Immunohistochemistry and real-time PCR were used to detect the angiotensin-converting enzyme（ACE）,ACE2 protein and mRNA in each group. The thiobarbituric acid method was used to detect the content of malondialdehyde（MDA） in each group. The mRNA of MCP-1,ICAM-1,TGF-β,and TNF-α were detected by real-time PCR in each group. Results Compared with the normal control group,diabetic mice had glomerular hypertrophy,increased mesangial matrix,and mesangial cell proliferation in the renal tissues. The above situation was alleviated in the kidney tissues of the jaundice group and the benazepril group. Compared with the normal control group,the relative expression of Ang II,α-SMA and Collagen IV increased in the kidney tissues of the diabetic group（all P〈0. 05）. Compared with the diabetic group,the relative expression of Ang II,α-SMA,and Collagen IV decreased in the kidney tissues of the ast

关 键 词：黄芪 糖尿病 糖尿病肾病 肾素-血管紧张素系统 血管紧张素Ⅱ Α-平滑肌肌动蛋白 Ⅳ型胶原 丙二醛 

分 类 号：R587.1[医药卫生—内分泌]