褪黑素抑制磷酸钙诱导的大鼠主动脉血管平滑肌细胞凋亡  

Melatonin inhibits apoptosis of rat aortic vascular smooth muscle cells induced by CaPO_4

作　　者：刘振华[1] 李步潆 段维勋[1] 金振晓[1] 俞世强[1] 

机构地区：[1]第四军医大学西京医院心脏外科,西安710032

出　　处：《中国体外循环杂志》2016年第2期112-115,119,共5页Chinese Journal of Extracorporeal Circulation

基　　金："十二五"国家科技支撑计划(2011BAI11B20);国家自然科学基金(81570231;81570230;81570232;81470415;81270170;81200151;81470411);陕西省国际科技合作与交流计划项目(2015KW-047);陕西省社会发展公关计划(2015SF104);陕西省科技统筹创新工程计划项目(2013KTCL03-01);陕西省自然科学基金(2014JM4106);西京医院学科助推计划(XJZT14203;XJGX13LC15;XJGX12C11)

摘　　要：目的观察褪黑素(Mel)对磷酸钙诱导的大鼠主动脉血管平滑肌细胞(SMCs)凋亡的影响。方法组织贴块法原代培养的血管SMCs随机分为3组:对照组(PBS)、磷酸钙(CaPO_4)组、磷酸钙+褪黑素(CaPO_4+Mel)组,Mel又按浓度分为0.1 mmol/L,0.5 mmol/L,1 mmol/L,2 mmol/L四组。CCK8法测定细胞增殖变化,TUNEL检测细胞凋亡,Western blot印迹法测定凋亡相关蛋白caspase3、Bcl-2、Bax表达。结果 CCK8法检测显示褪黑素可以抑制CaPO_4诱导的SMCs凋亡,促进其增殖,并具有时间和剂量依赖性。以1 mmol/L作用6 h显著。褪黑素作用6 h后,TUNEL检测显示CaPO_4诱导的SMCs凋亡明显减少(P<0.01),Western blot结果分析显示,与CaPO_4组比较,caspase3表达明显下降,同时,Bcl-2蛋白表达上调,Bax蛋白表达下调(P<0.01)。结论褪黑素可以抑制CaPO_4诱导的SMCs凋亡,其机制可能与下调caspase3和Bax蛋白,上调Bcl-2蛋白表达有关。Objective To investigate the effect of melatonin（Mel） on apoptosis of rat aortic vascular smooth muscle cells induced by CaPO4.Methods VSMCs were primarily cultured by tissue-sticking method and randomly divided into 3 groups： control group, CaPO4 group and CaPO4＋Mel group. According to the concentration of Mel,the CaPO4＋Mel groups were divided into 0.1 mmol/L,0.5 mmol/ L,1 mmol/ L,2 mmol/ L four groups. SMCs proliferation was determined by CCK8. Cells apoptosis were detected by TUNEL. The expressions of caspase 3、 Bcl-2 and Bax protein were investigated by Western blot.Results CCK8 method showed that Mel could inhibit SMCs apoptosis induced by CaPO4 and promoted SMCs proliferation, dependentof time and dose. Treatment with1 mmol / L Mel for 6h had the most significant inhibitive effect. After SMCs were treated with 1mmol/ L Mel for 6h,TUNEL demonstrated that the apoptosis rate of the SMCs significantly decreased（P〈0.01）. Western blot demonstrated caspase3 expression significantly decreased and Bcl-2 expression was up-regulated and Bax expression was down-regulated to compare to CaPO4group（P〈0.01）.Conclusion Mel can inhibit SMCs apoptosis induced by CaPO4,Its mechanisms may be relate to down-regulated the caspase3 and Bax protein expression and up-regulated the Bcl-2 protein expression.

关 键 词：褪黑素 平滑肌细胞 CaPO4诱导 细胞凋亡 

分 类 号：R543.16[医药卫生—心血管疾病]