摘要
目的 探讨晚期糖基化终产物 (advancedglycationendproducts ,AGEs)对体外培养乳鼠心肌细胞p5 7、p2 7、p2 1及hhLIM蛋白表达的影响。方法 采用酶消化法体外培养乳鼠心肌细胞 ,实验前换用含 1%胎牛血清的DMEM培养液饥饿培养 2 4h。应用Western蛋白印迹法检测不同浓度AGEs(5 0mg L、10 0mg L)对心肌细胞刺激 12、2 4、4 8h后细胞中p5 7、p2 7、p2 1及hhLIM蛋白表达情况。结果 不同浓度的AGEs刺激心肌细胞 12h均可使p2 7蛋白表达增加 ,以 5 0mg L浓度作用更为明显 ,刺激 2 4、4 8h无影响。AGEs对p5 7蛋白的表达呈动态变化 :刺激 12h时表达下降 ,2 4、4 8h呈增加趋势。p2 1蛋白在心肌细胞表达较弱 ,AGEs对其表达无显著影响。AGEs增加hhLIM蛋白的表达 ,以 5 0mg L浓度作用 12h最为显著。结论 AGEs通过增加心肌细胞细胞周期素依赖性激酶抑制因子p5 7和p2 7蛋白及hhLIM蛋白的表达 ,参与心室重塑的发生和发展。
Objective To investigate the effects of advanced glycosylation end-products (AGEs) on the expression of cyclin-dependent kinase inhibitors(p57, p27, p21) and hhLIM in neonatal rat cardiac myocytes (CMs). Methods CMs were cultured for 3～5 days in vitro using trypsin digestion method. After serum starving for 24 hours, the cells were exposed to AGEs (50 mg/L、100 mg/L) for 12, 24 and 48 hours respectively and western blotting was used to detect the expression of p57, p27, p21 and hhLIM.Results The expression of p27 was increased 12 hours after treatment with AGEs at different concentrations,with the effect of AGEs at 50 mg/L being more obvious,while the expressions at 24 hours and 48 hours after treatment were not significant. The expression of p57 after AGEs stimulation was fluctuant, in that 12 hours after treatment the expression of p57 showed a downward trend while 24 hours and 48 hours after treatment it presented an upward trend. The expression of p21 in CMs was weak and the effect of AGEs on it was not obvious. AGEs could increase the expression of hhLIM at different time points, especially at 12 h after treatment with 50 mg/L AGEs.Conclusions AGEs increase the expressions of cyclin-dependent kinase inhibitors p57 and p27 gene and increase the expression of hhLIM gene, which may play a role in cardiac ventricular remodeling.
出处
《中华老年心脑血管病杂志》
CAS
2004年第6期399-402,共4页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
基金
广东省自然科学基金研究团队项目 (10 717)
国家重点基础研究发展规划 (G2 0 0 0 0 5 690 5 )
