山松醇同源物1在缺氧/复氧诱导的人肾小管上皮细胞损伤中的作用及机制研究  

Role and mechanism of pumilio homolog 1 in hypoxia/reoxygenation induced injury of human renal tubular epithelial cells

作　　者：胡圣国 郭义 袁超[1] 李又空[1] 王敏[2] 朱敏 Hu Sheng-guo;Guo Yi;Yuan Chao;Li You-kong;Wang Min;Zhu Min(Department of Urology,Jingzhou Hospital Affiliated to Yangtze University,Jingzhou 434020,China;Department of Urology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]长江大学附属荆州医院泌尿外科,荆州434020 [2]武汉大学人民医院泌尿外科,武汉430060

出　　处：《临床肾脏病杂志》2024年第10期841-846,共6页Journal Of Clinical Nephrology

基　　金：湖北省自然科学基金青年项目(2019CFB302)。

摘　　要：目的探究山松醇同源物1(pumilio homolog 1,PUM1)在缺氧/复氧(hypoxia/reoxy-genatio,H/R)诱导的人肾小管上皮细胞(human renal tubular epithelial cells,HK-2)损伤中的作用及机制。方法体外建立HK-2细胞的H/R模型;通过小干扰RNA(small interfering RNA,siRNA)敲低HK-2细胞的PUM1表达;细胞等量随机法分为对照(Control)组、H/R组、H/R+siRNA组。蛋白质印迹法用以检测PUM1蛋白表达水平;细胞计数试剂盒8用以检测细胞活力;过氧化氢(hydrogen peroxide,H2O2)、丙二醛(malondialdehyde,MDA)和超氧化物歧化酶(superoxide dismutase,SOD)评估氧化应激水平;流式细胞术用以检测细胞凋亡水平。结果与Control组相比,H/R 3 h组(1.76±0.11比0.98±0.05)、H/R 6 h组(2.89±0.14比0.98±0.05)、H/R 12 h组(3.78±0.08比0.98±0.05)PUM1的蛋白表达水平随着缺氧时间的延长逐渐增高(均P<0.05)。与H/R组相比,H/R+si-PUM1组敲低PUM1的表达能够显著改善H/R后HK-2细胞的细胞活力(73.67±3.42比29.60±2.94)、氧化应激[H_(2)O_(2):(13.53±0.85)μmol/L比(22.43±1.12)μmol/L、MDA:(16.03±0.70)μmol/L比(31.20±1.50)μmol/L、SOD:(34670±1800)U/L比(5730±1220)U/L]及凋亡水平[(14.89±1.65)%比(39.71±1.94)%](均P<0.05)。结论PUM1在H/R诱导的HK-2细胞中上调,抑制PUM1的表达可以减少氧化应激及细胞凋亡水平,从而减轻H/R损伤。Objective To explore the role and mechanism of pumilio homolog 1(PUM1)in hypoxia/reoxygenation induced cell injury in human renal tubular epithelial cells(HK-2).Methods A hypoxia/reoxygenation(H/R)model of HK-2 cells was established in vitro.PUM1 expression was knocked down through small interfering RNA(siRNA).Cells were randomized into three groups of control,hypoxia/reoxygenation(H/R)and H/R+siRNA.Western blot(WB)method was used for detecting the expression level of PUM1 protein.Cell Count Kit 8(CCK-8)was employed for detecting cell viability.Hydrogen peroxide(H2O2),malondialdehyde(MDA)and superoxide dismutase(SOD)were used for evaluating the levels of oxidative stress.Flow cytometry was utilized for detecting the level of cell apoptosis.Results As compared with control group,protein expression level of PUM1 in H/R 3 h group(1.76±0.11 vs 0.98±0.05),H/R 6 h group(2.89±0.14 vs 0.98±0.05)and H/R 12 h group(3.78±0.08 vs 0.98±0.05)gradually spiked with the prolongation of hypoxic time.As compared with H/R group,knocking down the expression of PUM1 significantly improved the cell viability(73.67±3.42 vs 29.60±2.94),oxidative stress[H_(2)O_(2):(13.53±0.85)μmol/L vs(22.43±1.12)μmol/L,MDA:(16.03±0.70)μmol/L vs(31.20±1.50)μmol/L,SOD:(34670±1800)U/L vs(5730±1220)U/L]and apoptotic level[(14.89±1.65)%vs(39.71±1.94)%]after H/R in H/R+si-PUM1 group.Conclusion PUM1 is up-regulated in H/R induced HK-2 cells and its inhibition may alleviate H/R injury through reducing oxidative stress and lowering cell apoptosis levels.

关 键 词：山松醇同源物1 人肾小管上皮细胞 缺氧/复氧 氧化应激 细胞凋亡 

分 类 号：R692[医药卫生—泌尿科学]