摘要
目的探讨醛固酮(Ald)对新生大鼠心脏成纤维细胞(CFs)内皮素(ET)表达的影响。方法采用胰酶消化法和差速贴壁分离法获取新生大鼠CFs,应用放射免疫法、免疫荧光检测法、RT-PCR方法分别测定不同条件下CFs培养液中ET水平和CFs中的ET-1含量以及ppET-1 mRNA的表达。结果醛固酮(10-9、10-8和10-7mol/L)可剂量依赖性地促进CFsppET-1mRNA的表达以及ET-1的合成和分泌,同时醛固酮(10-7mol/L)以时间依赖的方式促进CFsppET-1 mRNA的表达,在作用2h后表达开始增加,4h达高峰,以后逐渐降低。提前给予螺内酯(10-6mol/L)能抑制醛固酮(10-7mol/L)的上述作用。结论醛固酮能促使CFsppET-1 mRNA的表达以及ET-1的合成和分泌,且此作用可能是通过盐皮质激素受体介导。
Objective To investigate the effects of aldosterone on the expression of endothelin (ET) in cultured neonatal rat cardiac fibroblasts (CFs). Methods CFs were isolated by trypsin digestion. ET concentration in conditioned medium was measured by radioimmunoassay, intracellular ET-1 level was evaluated by immunofluorescence assay, and the expression of preproendothelin-1 (ppET-1) was detected using reverse transcriptase-polymerase chain reaction (RT-PCR) method. Results Aldosterone ( 10^-9 10 ^-8, 10^-7 mol/L) induced a dose-dependent changes in ppET-1 mRNA expression, as well as ET-1 synthesis and secretion in CFs. Meanwhile, aldosterone ( 10^-7 mol/L) time-related induced ppET-1 mRNA expression in CFs, which began to increase in 2 h and reached the highest level in 4 h, thereafter decreased. The effects of aldosterone ( 10^-7 mol/L) were significantly inhibited by the pre-incubation with spironolactone (10^-6 tool/L). Conclusion Aldosterone increases the expression of ppET-1 mRNA, ET-1 synthesis and secretion via mineralocorticoid receptor.
出处
《基础医学与临床》
CSCD
北大核心
2008年第1期35-39,共5页
Basic and Clinical Medicine
基金
河北省教育厅基金(ZH2006004)
关键词
醛固酮
螺内酯
内皮素
心脏成纤维细胞
aldosterone
spironolactone
endothelin
cardiac fibroblasts