血清饥饿条件下δ阿片受体激活对心肌细胞存活的影响  

EFFECTS OF δ-OPIOID RECEPTOR STIMULATION ON SURVIVAL OF CULTURED MYOCARDIAL CELLS UPON TO SERUM DEPRIVATION

作　　者：王大鹏[1] 王洪新[1] 唐博[1] 喻晓春[2] 

机构地区：[1]辽宁医学院药理教研室,辽宁锦州121001 [2]中国中医科学院实验中心,北京100700

出　　处：《中国应用生理学杂志》2008年第3期274-278,共5页Chinese Journal of Applied Physiology

基　　金：辽宁省自然科学基金(20042170)

摘　　要：目的:研究血清饥饿对心肌造成损伤过程中,δ阿片受体激活是否对心肌细胞的存活起保护作用。方法:体外原代培养乳大鼠心肌细胞。结晶紫法测定心肌细胞活力;流式细胞仪检测细胞周期S+G2+M百分比;流式细胞仪检测心肌细胞的凋亡指数;Western蛋白印迹法检测心肌细胞胞浆内caspase-3蛋白表达。结果:体外原代培养乳大鼠心肌细胞经无血清培养48小时后,心肌细胞出现明显的凋亡,caspase-3表达增加;当给予不同浓度的δ阿片受体激动剂[D-Ala2,D-Leu5]脑啡肽(DADLE)作用于心肌细胞后,能明显抑制心肌细胞凋亡,并且0.1μmol.L-1DADLE的作用最为显著,表现为,心肌细胞活力增加,心肌细胞的S+G2+M百分比升高;心肌细胞的凋亡指数下降,caspase-3蛋白表达降低;0.1μmol.L-1DADLE对心肌细胞的保护作用能被δ阿片受体拮抗剂naltrindole10μmol.L-1完全阻断,表现为,心肌细胞活力降低,心肌细胞的凋亡指数升高;心肌细胞的S+G2+M百分比降低;caspase-3的蛋白表达明显升高;说明DADLE通过激活δ阿片受体对心肌细胞起保护作用。结论:在血清饥饿条件下,δ阿片受体激活对心肌细胞的存活有保护作用。Ain： To study upon to serum deprivation if δ-opioid receptor activation has direct effect on cultured impaired cardiomyocytes survival. Methods： Myocardial cells of neonatal rats were cultured in vitro. The cell viability was determined with crystal violet staining uptake. The percentage of S ＋ G2 ＋ M in cell cycle was determined by flow cytometry. Apoptosis rates were determined by flow cytometry（FCM）. The expression of Caspase-3 were investigated by Western blotting. Results： Myocardial cells of neonatal rats were cultured of serum-free in vitro,apoptotic index was significantly increased, the expression of Caspase-3 was significantly increased, free-serum induced apoptosis in cardiac myoeytes after 48 h. At concentrations of 10 nmol· L^-1- 10 μmol·L^-1, a delta opoid receptor agonist [ D-Ala2 , D-Leu5 ]-enkephalin DADLE promoted the myocardial cells survival, in a concentration-dependent manner. The optimal response was achieved at 0.1 μmol·L^-1 , which increase survival index of cardiac myocyte, percentage of S ＋ G2 ＋ M in cell cycle,decrease apoptotic index of cardiac myocyte, and the expression activate caspase-3.δ-opioid receptor antagonist nahrindole at 10 μmol·L^-1 inhibited the promoting effects of DADLE, which decrease survival index of cardiac myocyte, and percentage of S ＋ G2 ＋ M in cell cycle, increase apoptotic index of cardiac myocyte and the expression of Caspase-3. Conclusion： The protective of δ-opioid re ceptor activation can promote survival in cultured impaired myocardial cells.

关 键 词：心肌细胞 Δ阿片受体 [D-Ala2 D-Leu5]脑啡肽 NALTRINDOLE 细胞存活 CASPASE-3 

分 类 号：R972[医药卫生—药品]