摘要
目的:探讨人胎儿心脏成纤维细胞(FCFS)经不同浓度的醛固酮(ALD)干预后增殖的变化以及对细胞中I、III型胶原表达的影响。方法:采用胶原酶II消化法、差速贴壁法、差速脱壁法获取并纯化FCFS,在不同浓度醛固酮作用下,应用CCK-8活细胞计数试剂盒检测细胞的增殖率。应用RT-PCR方法检测细胞中I型胶原前胶原A1(COL1A1)和III型胶原前胶原A1(COL3A1)mRNA的变化,应用Western blot检测COL1A1、COL3A1蛋白表达的变化。结果:ALD可浓度依赖性促-进FCFS增殖;低浓度的ALD(10-9、10-8、10-7mol/L)可明显促进FCFS中COL1A1、COL3A1表达,而10-6mol/L的ALD没有明显作用,10-5mol/L的ALD甚至起到抑制作用。结论:ALD可促进FCFS的细胞增殖,其增殖程度随浓度升高而升高;ALD在一定浓度范围内可以促进COL1A1和COL3A1表达,高浓度ALD对两种分子表达具有抑制作用。
AIM：To investigate the effect of aldosterone（ALD） on the proliferation of fetal cardiac fibroblasts（FCFS） and the production of collagen I and collagen III in FCFS.METHODS： FCFS were isolated by collagenase II and purified with differential attachment and detachment method.The proliferation of FCFS after ALD administration was assessed by CCK-8.The mRNA expression of COL1A1 and COL3A1 were assessed by reverse transcription polymerase chain reaction（RT-PCR） and the protein production of COL1A1 and COL3A1 were assessed by Western blot.RESULTS： ALD facilitated the proliferation of FCFS concentration-dependently.ALD with lower concentration（10-9,10-8,10-7 mol/L） significantly improved the expression of COL1A1 and COL3A1,while ALD with higher concentration of had no obvious effect.CONCLUSION： ALD improved the proliferation of FCFS concentration-dependently.And in a certain concentration range,ALD improved the expression of COL1A1 and COL3A1 while higher concentration had opposite effect.There is no linear relationship among the effects of ALD on the proliferation of FCFS,expression and protein production of COL1A1 and COL3A1.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2011年第4期386-388,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助项目(30872065)
