基于PPARγ/ERK1/2信号通路探讨补肾开玄通络方对db/db小鼠的肾脏保护作用  

Exploration on the renal protective effect of Bushen Kaixuan Tongluo Formula on db/db mice based on PPARγ/ERK1/2 signaling pathway

作　　者：王友 张程斐 周洛卿 贾晓蕾 秦灵灵[1] 王海焱[1] 吴利佳 黄瑜坤 包新翠 杨靖 吕翠岩[3] 刘铜华[1] WANG You;ZHANG Chengfei;ZHOU Luoqing;JIA Xiaolei;QIN Lingling;WANG Haiyan;WU Lijia;HUANG Yukun;BAO Xincui;YANG Jing;LYU Cuiyan;LIU Tonghua(Beijing University of Chinese Medicine,Beijing 100029,China;Beijing University of Chinese Medicine Third Affliated Hospital,Beijing 100029,China;Beijing Hospital of Traditional Chinese Medicine,Capital Medical University,Beijing 100010,China)

机构地区：[1]北京中医药大学,北京100029 [2]北京中医药大学第三附属医院,北京100029 [3]首都医科大学附属北京中医医院,北京100010

出　　处：《中华中医药杂志》2024年第11期5849-5855,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：教育部、科技部2020年度新建高等学校学科创新引智基地项目(No.B20055)。

摘　　要：目的:观察补肾开玄通络方对db/db小鼠肾脏的保护作用及机制。方法:30只db/db小鼠根据空腹血糖随机分为5组:模型组,厄贝沙坦组,补肾开玄通络方低、中、高剂量组,每组6只,并设6只m/m小鼠为正常组。每2周检测各组小鼠体质量。连续给药12周后取材,测定空腹血糖、尿白蛋白、尿肌酐、尿白蛋白肌酐比值(ACR)、血肌酐(Scr)、尿素、血清总胆固醇(TC)、甘油三酯、胰岛素(INS)、天门冬氨酸氨基转氨酶、丙氨酸氨基转移酶水平,取肾脏包埋切片行HE染色、PAS染色及Masson染色。免疫组化法和RT-qPCR检测肾脏中PPARγ、ERK1/2蛋白和mRNA表达。结果:与模型组比较,补肾开玄通络方各剂量组小鼠体质量显著降低(P<0.05),补肾开玄通络方中、高剂量组小鼠尿液ACR水平降低(P<0.01,P<0.05),补肾开玄通络方中剂量组小鼠Scr、TC显著降低(P<0.05,P<0.01),补肾开玄通络方各剂量组INS含量均显著降低(P<0.01)。补肾开玄通络方各剂量组可减轻肾小球内毛细血管扩张、基底膜增厚和肾小管上皮细胞的变性、坏死,减轻糖原沉积,改善肾脏纤维化水平。免疫组化检测发现,与模型组比较,补肾开玄通络方各剂量组PPARγ蛋白水平显著升高(P<0.01),ERK1/2蛋白水平显著降低(P<0.01)。RT-qPCR检测发现,与模型组比较,补肾开玄通络方中、高剂量组PPARγmRNA水平显著升高(P<0.05,P<0.01),ERK1 mRNA水平显著降低(P<0.05,P<0.01),各剂量组ERK2 mRNA水平显著降低(P<0.01,P<0.05)。结论:补肾开玄通络方可改善糖尿病肾病小鼠的肾功能,降低尿蛋白,减轻肾脏损伤,其作用机制可能与PPARγ/ERK1/2信号通路有关。Objective:To observe the protective effect and mechanism of Bushen Kaixuan Tongluo Formula(BKTF)on the kidney of db/db mice.Methods:Thirty db/db mice were randomly divided into five groups based on fasting blood glucose levels:the model group,the irbesartan group,the low-dose,the medium-dose and the high-dose BKTF groups,six in each group,and six m/m mice were included as the normal group.Every 2 weeks,the body weight of mice in each group was measured.After 12 weeks of continuous administration,specimens were collected for measurement of fasting blood glucose,urinary albumin,urinary creatinine,urinary albumin creatinine ratio(ACR),serum creatinine(Scr),urea,serum total cholesterol(TC),triglycerides,insulin(INS),aspartate aminotransferase,and alanine aminotransferase levels.Kidney sections were embedded and subjected to staining with Hematoxylin-Eosin(HE),Periodic Acid-Schiff(PAS),and Masson’s trichrome.Immunohistochemical and RT-qPCR detected PPARγand ERK1/2 protein and mRNA expression in the kidney.Results:Compared with the model group,the body weight of mice in the each dose BKTF group was significantly reduced(P<0.05),the urine ACR level of mice in the mediumand high-dose BKTF groups was reduced(P<0.01,P<0.05),the Scr and TC level of mice in the medium-dose BKTF group was reduced(P<0.05,P<0.01),the INS content of all dose BKTF groups was reduced(P<0.01).All doses BKTF groups could can alleviate the dilation of capillaries in the glomerulus,thickening of the basement membrane,and degeneration and necrosis of renal tubular epithelial cells,reduce glycogen deposition,and improve the level of renal fibrosis.Immunohistochemical detection showed that compared with the model group,the group of all dose BKTF groups had the levels of PPARγprotein signifcantly increased(P<0.01),while the levels of ERK1/2 protein decreased(P<0.01).RT-qPCR detection showed that compared with the model group,the group of medium-and high-dose BKTF groups had the levels of PPARγmRNA signifcantly increased(P<0.05,P<0.01),while the level

关 键 词：补肾开玄通络方 糖尿病肾病 DB/DB小鼠 PPARΓ ERK1/2 

分 类 号：R285.5[医药卫生—中药学]