缺氧诱导因子-1调控血红素氧化酶-1在急性肾损伤的作用机制  

The mechanism of Heme oxygenase-1 regulated by Hypoxia-inducible factor-1 in acute kidney injury

作　　者：李金岩 李柏均 胡伟民 邹凡 余伟民[1] 程帆[1] Li Jinyan;Li Bojun;Hu Weimin;Zhou Fan;Yu Weimin;Cheng Fan(Department of Urology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院泌尿外科,武汉430060

出　　处：《中华实验外科杂志》2024年第1期74-77,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探究缺氧诱导因子-1(HIF-1)在肾缺血再灌注损伤中引起急性肾损伤(AKI)的作用及机制的研究。方法C57小鼠20只(6~8周龄,体重20~25 g),其中10只为野生型(WT)鼠,10只为纯合子肾小管上皮细胞特异敲低HIF-1(cKO)小鼠。按照简单随机法进行分组,将WT小鼠随机分为假手术(Sham)组和缺血再灌注(IR)组,每组各5只;cKO小鼠分随机为Sham组与IR组,每组各5只。用1%戊巴比妥钠注射小鼠尾静脉,手术组分离两侧肾动静脉结扎60 min后解除夹闭,24 h后切除肾脏,构建肾缺血再灌注模型。通过苏木精-伊红(HE)染色判断肾小管损伤情况;酶联免疫吸附测定(ELISA)检测肾脏组织中谷胱甘肽(GSH)、丙二醛(MDA)和铁含量;蛋白质印迹法(Western blot)检测血红素氧化酶1(HO-1)及铁死亡相关指标的蛋白表达水平;免疫组织化学染色(IHC)检测HIF-1、HO-1、铁死亡相关组织的表达。两组间差异比较采用单样本t检验。结果HE染色结果显示,WT小鼠中IR组肾小管上皮细胞受损程度高于Sham组(9.78±1.07比2.43±0.93,t=5.438,P<0.05);敲低HIF-1后cKO-IR组细胞损伤程度也高于WT-IR组(28.59±2.57比9.78±1.07,t=3.348,P<0.05),差异均有统计学意义。Western blot结果显示在WT小鼠中IR组HIF-1的表达趋势高于Sham组(0.996±0.068比0.634±0.072,t=10.482,P<0.05),IR组HO-1的表达趋势也高于Sham组(0.734±0.153比0.592±0.097,t=8.942,P<0.05),差异均有统计学意义。免疫组织化学染色结果显示,WT小鼠与cKO小鼠在IR模型中比较,WT-IR组中HIF-1阳性表达面积高于cKO-IR组(8.52±0.21比4.35±0.16,t=5.769,P<0.05),WT-IR组中HO-1阳性表达面积高于cKO-IR组(8.53±0.22比3.43±0.18,t=7.534,P<0.05),差异均有统计学意义。结论肾缺血再灌注模型中HIF-1表达反应性增加,敲低HIF-1可加重肾损伤,HIF-1可能通过调控HO-1缓解铁死亡进而减轻肾损伤。Objective This study investigates the role and mechanism of hypoxia-inducible factor-1(HIF-1)in acute kidney injury(AKI)induced by renal ischemia-reperfusion(IR).Methods A total of 20 C57 mice,aged 6-8 weeks and weighing 20-25 g.This group included 10 wild-type(WT)mice and 10 mice with tubular epithelial cell-specific knockdown of HIF-1(cKO).The mice were randomly assigned to either a sham operation(Sham)group or an IR group,with 5 mice in each subgroup.1%pentobarbital sodium was administered intravenously for anesthesia.In the surgical procedure,the renal arteries and veins were ligated for 60 minutes and then released.Kidneys were harvested 24 hours later to establish the ischemia-reperfusion model.Renal tubular damage was evaluated using hematoxylin-eosin staining(HE).Glutathione(GSH),malondialdehyde(MDA),and iron levels in kidney tissue were determined by enzyme-linked immunosorbent assay(ELISA).Western blotting was employed to assess heme oxygenase-1(HO-1)protein expression and proteins related to iron-induced cell death.Immunohistochemistry(IHC)was utilized to detect the expression of HIF-1,HO-1,and proteins associated with iron-induced cell death.All data were presented as mean±standard deviation,and group comparisons were made using an independent t-test.Results HE staining indicated that the IR group of WT mice exhibited more severe renal tubular epithelial cell damage compared to the Sham group(9.78±1.07 vs.2.43±0.93,t=5.438,P<0.05).In the cKO mice,cell damage in the IR group was more severe than in the WT-IR group after HIF-1 knockdown(28.59±2.57 vs.9.78±1.07,t=3.348,P<0.05),showing statistically significant differences.Western blotting results revealed higher expression of HIF-1 and HO-1 in the IR group of WT mice compared to the Sham group(HIF-1:0.996±0.068 vs.0.634±0.072,HO-1:0.734±0.153 vs.0.592±0.097,both P<0.05).Immunohistochemistry findings showed that in the WT-IR group,both HIF-1 and HO-1 expressions were significantly higher compared to the cKO-IR group(HIF-1:8.52±0.21 vs.4.35±0.16

关 键 词：缺氧诱导因子-1 缺血再灌注 血红素氧化酶1 铁死亡 

分 类 号：R692[医药卫生—泌尿科学]