褪黑素通过p38／MAPK信号通路调控oxLDL诱导的人脐静脉内皮细胞MLCK的表达  

The expression of myosin light chain kinase in oxLDL-induced human umbilical vein endothelial cell is regulated by the melatonin through the pathway of p38/MAPK

作　　者：肖林林[1] 朱华庆[1] 程筱雯[1] 江志奎[1] 左莉[1] 周青[1] 桂淑玉[1] 汪渊[1] 

机构地区：[1]安徽医科大学分子生物学实验室,生物化学教研室和安徽省/省部共建教育部重要遗传病基因资源利用重点实验室,合肥230032

出　　处：《安徽医科大学学报》2008年第1期16-20,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金资助项目(编号:30570750);安徽省自然科学基金(编号:070413079);安徽省教育厅基金(编号:KJ2007B275)

摘　　要：目的探讨褪黑素(MLT)通过p38/MAPK信号通路调控oxLDL诱导的人脐静脉内皮细胞肌球蛋白轻链激酶(MLCK)的表达。方法体外培养人脐静脉内皮细胞(HU-VEC),分空白对照组、DMSO对照组、DMSO+oxLDL组、DM-SO+oxLDL+SB203580组、DMSO+oxLDL+MLT组、DMSO+oxLDL+MLT+SB203580组。RT-PCR、Western blot、γ-32P-ATP掺入法分别检测各组HUVEC MLCK转录、表达和活性及MLT对p38磷酸化的影响。结果RT-PCR显示oxLDL组MLCK转录增强,MLT组与SB203580组均抑制MLCK的转录;Western blot结果表明oxLDL组MLCK表达、p38磷酸化增强,MLT组与SB203580组均抑制MLCK的表达及p38磷酸化;γ-32P-ATP掺入法检测MLCK活性提示oxLDL组MLCK活性增强,MLT与SB203580降低MLCK活性。结论MLT可能通过p38/MAPK通路调控MLCK的转录、表达和活性。Objective To study the possibility of the expression of myosin light chain kinase in oxLDL-induced human umbilical vein endothelial cell is regulated by the melatonin through the pathway of p38/MAPK. Methods HUVEC were cultured in vitro. Treatments for the 6 groups were ： blank, DMSO, DMSO ＋ oxLDL, DMSO ＋ oxLDL ＋ SB203580, DMSO ＋ oxLDL ＋ MLT, DMSO ＋ oxLDL ＋ MLT ＋ SB203580. RT-PCR, Western blot, γ-^32p-ATPincor poration into myosin light chain were used to evaluate the influence of MLT and p38 inhibitor SB203580 on oxLDL induced HUVEC MLCK expression and p38 phosphorylation. Results RT-PCR revealed that MLCK transcription was significantly increased under oxLDL induction while significantly inhibited under MLT and SB203580, Western blot assay detected increased p38 phosphorylation under induction of oxLDL while decreased under MLT and SB203580,γ-^32p-ATP incorporation into myosin light chain revealed MLCK activity was significantly increased under oxLDL, but decreased under MLT and SB203580. Conclusion MLT may control the expression and actibity of MLCK in oxLDL-induced human umbilical vein endothelial cell via p38/MAPK pathway.

关 键 词：肌球蛋白轻链激酶 褪黑激素 磷酰化 脐静脉/细胞学 

分 类 号：R345[医药卫生—基础医学] R977.1