缺氧诱导因子-1α抑制剂YC-1改善糖尿病肾病小鼠肾脏损伤的机制研究  

Renal protective effect and mechanism research of hypoxia inducible factor-1αinhibitor YC-1 in diabetic nephropathy mice

作　　者：贾君杰 邢海帆 张群子 刘奇烨 汪年松[1] 范瑛[1] JIA Junjie;XING Haifan;ZHANG Qunzi;LIU Qiye;WANG Niansong;FAN Ying(Department of Nephrology,Shanghai Sixth People's Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200233,China)

机构地区：[1]上海交通大学医学院附属第六人民医院肾内科,上海200233

出　　处：《上海交通大学学报（医学版）》2023年第9期1089-1098,共10页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(81870468,82170727);上海交通大学医学院“双百人”项目(20191833);上海交通大学“交大之星”医工交叉重点项目(YG2023ZD21)。

摘　　要：目的·研究缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)抑制剂YC-1对小鼠糖尿病肾病(diabetic nephropathy,DN)进展的影响及潜在机制。方法·将10周龄的雄性db/db小鼠(DN模型)和同窝野生型(WT)小鼠按是否给予YC-1分为4组,即WT组、WT+YC-1组、DB组、DB+YC-1组,每组6只。YC-1干预组予以YC-1(20 mg/kg,1次/d)腹腔注射8周,非干预组同时予以等体积二甲基亚砜腹腔注射。干预8周后,检测小鼠血糖、体质量和肾脏质量,并收集血清、尿液、肾组织标本。检测小鼠血肌酐、尿白蛋白/肌酐比(urinary albumin-to-creatinine ratio,UACR)、尿中性粒细胞明胶酶相关脂质运载蛋白(neutropil gelatinase-associated lipocalin,NGAL)水平。肾脏行苏木精-伊红(H-E)染色、过碘酸-雪夫(PAS)染色观察组织病理损伤;马松(Masson)染色检测纤维化情况,免疫组织化学(免疫组化)法检测Ⅰ型胶原蛋白,Western blotting检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)水平;免疫组化法和Western blotting检测HIF-1α表达;TUNEL染色和Western blotting检测细胞凋亡水平;试剂盒检测肾脏超氧化物歧化酶(superoxide dismutase,SOD)活性和丙二醛(malondialdehyde,MDA)含量;Western blotting检测内质网应激(endoplasmic reticulum stress,ERS)标志物免疫球蛋白重链结合蛋白(immunoglobulin heavy chain binding protein,BiP;又称GRP78)、磷酸化蛋白激酶样内质网激酶(phospho-protein kinase R-like endoplasmic reticulum kinase,p-PERK)、总PERK、磷酸化真核起始因子2α(phospho-eukaryotic initiation factor 2α,p-eIF2α)、总eIF2α、激活转录因子4(activating transcription factor 4,ATF4)和C/EBP同源蛋白(C/EBP homologous protein,CHOP)的表达。结果·与WT组小鼠相比,DB组小鼠血糖升高,肾功能下降,肾脏病理损伤和纤维化加重,肾脏HIF-1α表达、氧化应激和ERS激活程度增加。与DB组小鼠相比,DB+YC-1组小鼠血糖无明显变化,但肾/体质量比、血肌酐、UACR、尿NGAL水�Objective·To investigate the effect of hypoxia inducible factor-1α(HIF-1α)inhibitor YC-1 on the progression of diabetic nephropathy(DN)in mice and the potential mechanism.Methods·Ten-week-old male db/db mice(DN model)and their nondiabetic wild-type(WT)littermates were divided into 4 groups(n=6)according to whether treated with YC-1 or not:WT group,WT+YC-1 group,DB group,and DB+YC-1 group.The treatment groups were intraperitoneally injected with YC-1(20 mg·kg−1)once a day,while the non-treatment groups received the same volumes of DMSO injection.After a total of 8 weeks of intervention,blood glucose,body weight,and kidney weight of all mice were measured.Serum,urine and kidney tissue samples were harvested.Serum creatinine,urinary albumin-to-creatinine ratio(UACR),and urine neutropil gelatinase-associated lipocalin(NGAL)levels were detected.The kidneys were stained with hematoxylin-eosin(H-E)and periodic acid-Schiff(PAS)to observe the pathological changes.Masson staining was used to detect fibrosis,collagen-Ⅰwas detected by immunohistochemistry,andα-smooth muscle actin(α-SMA)was detected by Western blotting.The expression of HIF-1αwas detected by both Western blotting and immunohistochemistry.TUNEL staining and Western blotting for apoptosis-related proteins were used to observe the cell apoptosis level.Superoxide dismutase(SOD)activity and malondialdehyde(MDA)level were detected by the kits.Endoplasmic reticulum stress(ERS)markers,including immunoglobulin heavy chain binding protein(BiP,also known as GRP78),phospho-protein kinase R-like endoplasmic reticulum kinase(p-PERK),total PERK,phospho-eukaryotic initiation factor 2α(p-eIF2α),total eIF2α,activating transcription factor 4(ATF4),and C/EBP homologous protein(CHOP),were determined by Western blotting.Results·Compared with the WT group,the DB group showed significant rise of blood glucose,loss of renal function,severe kidney histopathology injuries and kidney fibrosis,increase of renal HIF-1αexpression,and aggravated oxidative stress and ERS.Whi

关 键 词：糖尿病肾病 缺氧诱导因子-1Α 氧化应激 内质网应激 

分 类 号：R587.2[医药卫生—内分泌]