参麦注射液通过下调miR-106a保护肾小管上皮细胞NRK-52E缺氧-复氧损伤的分子机制  

The molecular mechanism of Shenmai injection in protecting renal tubular epithelial cells(NRK-52E)from hypoxia and reoxygenation injury by down-regulating miR-106a

作　　者：黄真珍 王培琦 李冰 李晓媛 HUANG Zhen-zhen;WANG Pei-qi;LI Bing;LI Xiao-yuan(School of Pharmacy,Zhengzhou University,Zhengzhou 450000,Henan,China)

机构地区：[1]郑州大学药学院,郑州450000

出　　处：《医学研究生学报》2021年第5期476-481,共6页Journal of Medical Postgraduates

摘　　要：目的参麦注射液已被证实对心肌细胞缺血再灌注(I/R)损伤具有保护作用,但其对肾I/R损伤的保护作用并不明确。文章旨在探讨参麦注射液对肾小管上皮细胞NRK-52E缺氧-复氧损伤的的影响和可能分子机制。方法构建NRK-52E细胞缺氧复氧模型模拟肾I/R损伤过程。根据干预方法不同,将NRK-52E细胞分为正常对照组、I/R组(缺氧6 h-复氧24 h)、参麦+I/R组(参麦注射液处理30 min,缺氧6 h-复氧24 h)、miR-NC+I/R组(转染miR-NC,缺氧6 h-复氧24 h)、miR-106a+I/R组(转染miR-106a模拟物,缺氧6 h-复氧24 h)、anti-miR-NC+I/R组(转染anti-miR-NC,缺氧6 h-复氧24 h)、anti-miR-106a+I/R组(转染anti-miR-106a,缺氧6 h-复氧24 h)、参麦+miR-NC+I/R组(转染miR-NC,参麦注射液处理30 min,缺氧6 h-复氧24 h)、参麦+miR-106a+I/R组(转染miR-106a模拟物,参麦注射液处理30 min,缺氧6 h-复氧24 h)。细胞计数试剂盒(CCK-8)检测细胞活力,流式细胞术检测细胞凋亡,实时荧光定量PCR(RT-qPCR)检测miR-106a的表达水平,Western blot法检测磷脂酰肌醇-3-羟激酶/蛋白激酶B(PI3K/AKT)蛋白表达。结果与正常对照组Ki67蛋白(0.76±0.07)、细胞活性(0.83±0.05)比较,I/R组(0.33±0.02、0.35±0.02)显著降低(P<0.05),与正常对照组Cleaved-caspase-3蛋白表达(0.42±0.04)、凋亡率[(5.06±0.32)%]比较,I/R组[(0.86±0.07、(16.11±1.01)%]显著升高(P<0.05);与I/R组Ki67蛋白、细胞活性比较,参麦+I/R组(0.70±0.07、0.77±0.06)明显升高,与I/R组Cleaved-caspase-3蛋白表达、凋亡率比较,参麦+I/R组(0.54±0.04、7.24±0.53)明显降低(P<0.05)。I/R组NRK-52E细胞miR-106a的表达水平(3.27±0.25)较正常对照组(1.00±0.08)和参麦+I/R组(1.53±0.08)显著升高(P<0.05)。与miR-NC+I/R组比较,miR-106a+I/R组NRK-52E细胞miR-106a的表达水平显著升高,细胞活性、Ki67蛋白表达显著降低,细胞凋亡率、Cleaved-caspase-3蛋白表达显著升高(P<0.05);与anti-miR-NC+I/R组比较,anti-miR-106a+I/R组NRK-52E细胞miR-106a的�Objective Shenmai injection has been proved to have a protective effect on myocardial ischemia-reperfusion(I/R)injury,but its protective effect on renal I/R injury is not clear.The purpose of this study was to investigate the effect of Shenmai injection on renal tubular epithelial cells NRK-52E hypoxia-reoxygenation injury and its possible molecular mechanism.Methods NRK-52E cells hypoxia-reoxygenation model was constructed to simulate renal I/R injury.According to different intervention methods,NRK-52E cells were divided into normal control group,I/R group(hypoxia for 6 h,reoxygenation for 24 h),Shenmai+I/R group(Shenmai,hypoxia 6 h-reoxygenation 24 h),miR-NC+I/R group(transfected miR-NC,hypoxia 6 h-reoxygenation 24 h),miR-106a+I/R group(transfected miR-106a mimics,hypoxia 6 h-reoxygenation 24 h),anti-miR-NC+I/R group(transfected anti-miR-NC,hypoxia 6 h-reoxygenation 24 h),anti-miR-106a+I/R group(transfected anti-miR-106a,hypoxia 6 h-reoxygenation 24 h),Shenmai+miR-NC+I/R group(transfected miR-NC,Shenmai,hypoxia 6 h-reoxygenation 24 h),Shenmai+miR-106a+I/R group(transfected miR-106a mimics,Shenmai,hypoxia 6 h-reoxygenation 24 h).Cell viability was detected by cell counting kit(CCK-8),cell apoptosis was analysed by flow cytometry,the expression level of miR-106a was measured by real-time quantitative PCR(RT-qPCR),and the expression of phosphatidylinositol-3-hydroxykinase/protein kinase B(PI3K/AKT)protein was tested by Western blot.Results Compared with the normal control group,the cell viability(0.346±0.02 vs 0.833±0.05),p-PI3K(0.38±0.03 vs 0.70±0.07)and p-AKT protein(0.28±0.02 vs 0.60±0.05)expression of NRK-52E cells in I/R group were significantly decreased,and the apoptosis rate[(16.11±1.01)%vs(5.06±0.32)%]and miR-106a expression(3.27±0.25 vs 1.00±0.08)were significantly increased(P<0.05).Compared with I/R group,the cell viability(0.768±0.06 vs 0.346±0.02),p-PI3K(0.62±0.04 vs 0.38±0.03)and p-AKT protein(0.51±0.05 vs 0.28±0.02)expression of NRK-52E cells in Shenmai+I/R group were significantly i

关 键 词：参麦注射液 肾小管上皮细胞 miR-106a 缺氧-复氧损伤 

分 类 号：R-33[医药卫生]