血管紧张素Ⅱ及受体 2 诱导新生鼠肾小管上皮细胞凋亡的研究

Induced apoptosis of renal tubular cells from neonate rats by angiotensinⅡand AT2 receptor

关键词:  血管紧张素Ⅱ；CGP-42112A  凋亡

Key words:  angiotensinⅡ  AT₂ receptor  CGP-42112A  apoptosis

摘要:

［目的］研究血管紧张素Ⅱ和受体2对肾小管上皮细胞凋亡诱导的作用。 ［方法］ 取新生大鼠肾小管上皮细胞体外培养，传代后分别加入不同浓度的血管紧张素Ⅱ和AT₂受体激动剂CGP-42112A，孵育24 h, 流式细胞仪测定凋亡细胞比值, RT-PCR半定量检测AT₂受体mRNA表达。［结果］血管紧张素Ⅱ以10^-5mol/L、10^-6mol/L、10^-7 mol/L、10^-8mol/L浓度加入培养细胞24 h后，凋亡细胞比值分别为(21.73±1.25)%、(18.65±0.49)%、(17.29±0.67)%、(15.98±0.71)%，均显著增高于对照组(6.89±1.17)%，(P<0.001)。CGP-42112A以10^-5～10^-8mol/L浓度作用于细胞后凋亡细胞值分别为(22.32±2.69)%、(19.17±1.63)%、(17.98±1.96)%、(16.06±1.49)%，均显著高于对照组(7.04±0.61)%，(P<0.001)，凋亡细胞数与2种药物均呈剂量依赖性。RT-PCR半定量分析，两种药物作用于细胞后，随着浓度的逐渐增加，AT₂受体mRNA的表达均逐渐增强。 ［结论］ 血管紧张素Ⅱ和 AT₂ 受体兴奋剂均可诱导肾小管上皮细胞凋亡，血管紧张素Ⅱ诱导细胞凋亡可能通过AT₂ 受体途径实现。

Abstract:

［Objective］To invest the role of angiotensinⅡand AT₂ receptor in apoptosis of renal tubular cells. ［Methods］ In vitro neonate Wistarrat's renal tubular cells culture, AngiotensinⅡ(AngⅡ) or AT₂receptor agonist CGP-42112A in different doses were diffused separately, and at 24h, cells were analyzed by flow cytometry analysis of apoptosis, AT₂receptor mRNA expression was measured by RT-PCR. ［Results］In the groups with the diffusion of AngⅡ at 10^-5mol/L，10^-6mol/L，10^-7 mol/L and 10^-8mol/L , apoptotic cell rations were(21.73±1.25)%, (18.65±0.49)%, (17.29±0.67)% and (15.98±0.71)% respectively，being higher than that in group (6.89±1.17)%, (Ｐ＜0.001). In the groups with diffusion at 10^-5mol/L，10^-6mol/L，10^-7mol/L and 10^-8mol/L of CGP-42112A, apoptotic cell rastions were that (22.32±2.69)%,(19.17±1.63)%,(17.98±1.96)% and (16.06±1.49)% respectively, being higher than that of (7.04±0.61)%, ( Ｐ ＜0.001), a dose dependent manner was found, but difference was insignificant between AngⅡand CGP-42112A groups. RT-PCR results shown that the AT₂ receptor mRNA levels were up-regulated by both of CGP-42112A and AngⅡ. ［Conclusions］ AngiotensinⅡand CGP-42112A may up-regulated the renal tubular cells' apoptotic rations. The apoptosis was induced by AngiotensinⅡ maybe via the way of AT₂ receptor.