摘要
目的探讨12-脂氧化酶(12-LO)对系膜细胞血管紧张素(Ang)Ⅱ1型受体(ATlR)表达的影响。方法用AngⅡ刺激正常和12-LO基因敲除小鼠肾系膜细胞后,观察p38 MAPK活性和细胞外基质(ECM)蛋白的变化。用12-LO的作用产物12(S)-HETE刺激的系膜细胞、转染12-LO基因的系膜细胞和采用显微切割法从正常和12-LO基因敲除小鼠肾脏提取的肾小球来观察AT1R的表达。采用RT-PCR和Western印迹分别检测目标基因mRNA和蛋白的表达。结果AngⅡ的刺激可诱导正常系膜细胞p38 MAPK活性和ECM蛋白表达增高。然而,AngⅡ的刺激不能诱导12-LO基因敲除小鼠系膜细胞p38 MAPK活性和ECM蛋白表达升高。剂量依赖性和时间依赖性实验结果表明12(S)-HETE刺激可引起系膜细胞AT1蛋白水平增高,且AT1R mRNA水平升高有统计学意义(P＜0．01)。敲除肾小球内12-LO基因可有效地降低AT1R mRNA的表达(P＜0．01),转染12-LO基因至系膜细胞使AT1R蛋白和mRNA的表达明显增多(P＜0．01)。结论12-LO可上调系膜细胞AT1R的表达。
Objective To investigate the effect of 12-1ipoxygenase （12-LO） on the angiotensin Ⅱ type 1 receptor（AT1R） expression in mesangial cells （MC）. Methods p38 MAPK activation and ECM protein expression were determined using Ang Ⅱ-stimulated MC derived from normal and 12-LO knockout mice. AT1R expression was determined using 12-LO product 12（S）- HETE-stimulated MC, MC transfected with 12-LO gene and microdissected glomeruli derived from 12-LO knockout mice. RT-PCR and Western blot were used for evaluating mRNA and protein expression respectively. Results Ang Ⅱ stimulation increased p38 MAPK activation and ECM protein expression in normal MC, but not in MC derived from 12-LO knockout mice. Time-dependent and dose-dependent experiment showed that 12 （S）-HETE increased AT1R protein ＇ expression in MC. Similarly, 12（S）-HETE increased AT1R mRNA expression in MC compared with control MC （P 〈 0.01 ）. Furthermore, AT1R expression was lower in glomeruli derived from 12-LO knockout mice relative to genetic controls （P 〈 0.01） and MC stably overexpressing 12-LO had greater AT1R protein and mRNA expression relative to control MC （P 〈 0.01）. Conclusion 12-LO activation can upregulate ATIR expression in MC.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2006年第10期628-631,共4页
Chinese Journal of Nephrology
