吡哆胺和替米沙坦对人肾小管上皮细胞活性氧生成的影响

摘要

Objective The purpose of this study was to explore the protective effects of telmisartan and pyridoxamine on Reactive oxygen species(ROS)in cultured tubular epithelial ceIIs(HK-2). Methods Cultured HK-2 cells were divided into HK-2 control, angiotensin II (10-6 mol/1) group(Ang II ), telmisartan (telmisartan 10-6 mol/L+ Ang Ⅱ 10-6 mol/L, T) group, pyridoxamine group (pyridoxamine lmmol/L+ Ang Ⅱ 10-6 mol/L, Pl ), pyridoxamine group (pyridoxamine 10 mmol/L, P2), and T + P + angiotensin Ⅱ group (telmisartan 10-6 mol/L + pyridoxamine 10 mmol/L+Ang Ⅱ 10-6 mol/L, TP). ROS generation in cellular supernatant was detected by flow cytometry. Real-time quantitative PCR was performed to measure the mRNA level of expression of transforming growth factor-βl( TGF-β1) and receptor for advanced glycation end-products (RAGE). Results The level of ROS was reduced in PI , P2 and TP group (P < 0.01 ).There was no statistically difference between T groups and control (P > 0.05). Compared with the Ang II intervention, the mRNA expression level of RAGE and TGF-β1 were decreased in T, P and TP intervention (P < 0.05). Synergy decreased of TGF-β1 expression was showed in TP intervention (P < 0.01). Conclusion There is a synergistic effect of reducing HK-2 cells RAGE and TGF-β expression of HK-2 cells by combined use of telmisartan and pyridoxamine. However, pyridoxamine was effective than telmisartan in improving the function of oxidative stress.%目的 通过体外培养人肾小管上皮细胞株(HK-2)细胞,探讨吡哆胺和替米沙坦对肾小管上皮细胞活性氧生成影响.方法 HK-2细胞培养并按干预方式不同分为HK-2正常对照组,血管紧张素Ⅱ组(10-6 mol/L,AngⅡ组),替米沙坦组[替米沙坦(10-6mol/L)+ AngⅡ(10-6 mol/L),T组],吡哆胺1组[吡哆胺(1 mmol/L)+ AngⅡ(10-6 mol/L),P1组),吡哆胺2组[吡哆胺(10 mmol/L)+ AngⅡ(10-6 mol/L),P2],替米沙坦+吡哆胺联合组[替米沙坦(10-6 mol/L)+吡哆胺(10 mmol/L)+ AngⅡ(10-6 mol/L),TP组].流式细胞仪检测细胞上清液中的活性氧(ROS)浓度,荧光实时定量PCR方法检测HK-2细胞所表达的转化生长因子β1(TGF-β1)和晚期糖基化终产物受体(RAGE)mRNA表达量.结果 检测细胞上清液中的ROS浓度,与AngⅡ组比较,P1组和P2组均显著降低ROS浓度(P＜0.01),TP组的ROS生成也明显减少(P＜0.01),但T组与AngⅡ组间比较差异无统计学意义(P＞0.05)；与AngⅡ组比较,T组、P2组和TP组的HK-2细胞的RAGE,TGF-β1 mRNA表达量明显降低(P＜0.05)；与T组和P2比较,TP组HK-2细胞的TGF-β1 mRNA表达进一步降低(P＜0.01).结论 替米沙坦和吡哆胺可协同下调HK-2细胞转化生长因子TGF-β1和RAGE的表达；在改善氧化应激功能上,吡哆胺作用强于替米沙坦.