摘要
目的探讨缺氧诱导因子1α(HIF-1α)介导的信号通路在血管紧张素Ⅱ(AngⅡ)诱导肾间质纤维化(RIF)中的作用。方法体外培养肾小管上皮细胞,以不同浓度(10^-9~10^-6mol/L)的AngⅡ分别处理24h、48h时,用实时荧光定量PCR和Western印迹法分别检测肾小管上皮细胞中HIF-1α、脯氨酸羟化酶2(PHD2)及基质金属蛋白酶1组织抑制剂(TIMP-1)mRNA和蛋白的表达变化情况。结果随AngⅡ刺激浓度的增加,HIF。letmRNA的表达水平也增加,呈浓度依赖性。当AngⅡ浓度在10^-7mol/L,并且干预时间为24h时,HIF-1αmRNA的表达水平增加了166%。实时荧光定量PCR和Western印迹分析显示,相对于空白对照组,在AngⅡ干预的肾小管上皮细胞中HIF-1α、TIMP-1的mRNA和蛋白表达水平均升高(P〈0.05),而PHD2的mRNA和蛋白表达水平均下降(P〈0.05)。结论AngⅡ可能通过下调PHD2的表达而减少肾小管上皮细胞中HIF-1α的降解,从而上调HIF-1α及TIMP-1的表达,参与RIF。
Objective To explore the role of hypoxia inducible factor 1α (HIF-1α)- mediated signaling pathway in angiotensin Ⅱ (Ang Ⅱ ) induced renal' interstitial fibrosis. Methods Renal tubular epithelial cells were cultured and treated with different concentrations (10^-9-10^-6 mol/L) of Ang Ⅱ for 24 h and 48 h. Real-time quantitative PCR and Western blotting were preformed to detect the mRNA and protein expressions of HIF-1α, prolyl hydroxylase 2 (PHD2) and tissue inhibitor of metalloproteinase 1 (TIMP-1) in renal tubular epithelial cells. Results HIF-1α mRNA level was increased with Ang Ⅱ treatment in a concentration dependent manner. When ceils were treated with Ang Ⅱ concentration at 10^-7 mol/L for 24 h, the mRNA level was markedly increased by 166%. Furthermore, by real-time quantitative PCR and Western blotting, compared with the control group, AngⅡ increased the mRNA and protein levels of HIF-1α and TIMP-1 (P〈 0.05, respectively), while the mRNA and protein levels of PHD2 were decreased markedly (P〈 0.05, respectively) in renal tubular epithelial cells. Conclusion Ang Ⅱ reduces HIF-1α degradation in renal tubular epithelial ceils probably by reducing the expression of PHD2, which increases the expressions of HIF-1α and TIMP-1 involved in renal interstitial fibrosis.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2011年第3期194-197,共4页
Chinese Journal of Nephrology