吗啡对血清饥饿致培养乳大鼠心肌细胞凋亡的影响及机制

目的利用体外原代培养的乳大鼠心肌细胞,研究吗啡对血清饥饿诱导心肌细胞凋亡的影响及其机制。方法体外原代培养乳大鼠心肌细胞,各组分别给药作用48h后,用MTT法测心肌细胞的活力;用Annexin V-FITC/PI双标记法测心肌细胞的凋亡率;用流式细胞仪分析心肌细胞周期;用Westernblot法测PKC和Caspase-3蛋白的表达。结果体外原代培养的乳大鼠心肌细胞经无血清饥饿作用48h后,心肌细胞可出现明显的凋亡现象;给予1μmol·L-1吗啡作用于心肌细胞后,心肌细胞的这种凋亡现象可明显被抑制,表现为:心肌细胞凋亡率降低、Caspase-3蛋白表达减少;但吗啡的这种抑制心肌细胞凋亡作用可被10μmol·L-1 naloxone完全阻断;并且给予10μmol·L-1GF109203X或1μmol·L-1Staurosporine与吗啡共同作用时,吗啡抑制心肌细胞凋亡的作用亦在一定程度上降低,表现为:心肌细胞凋亡率增加、Caspase-3蛋白表达增加、PKC蛋白表达减少。结论吗啡可激活心肌细胞膜上的阿片受体,通过PKC途径对血清饥饿诱导的心肌细胞凋亡具有抑制作用。

Effect of morphine on serum hungry-induced apoptosis in cardiac myocytes

Aim Myocardial cells of neonatal rats were cultured in vitro to study aim of morphine on serum hungry-induced apoptosis in cardiac myocytes and its mechanism.Methods Myocardial cells of neonatalrats were cultured in vitro.48 hours later,different agents were added to cardiac myocytes.The cellular survival was determined with MTT colormeteric assay;apoptosis rates were determined by Annexin V-FITC/PI;cell cycle was determined by flow cytometry;Caspase-3 and PKC were investigated by Western blot.Results Free-serum induced apoptosis in cardiac myocytes was shown after 48 hours;morphine(1 μmol·L-1)could inhibit apoptosis in cardiac myocytes,manifestation,apoptosis rates were decreased,Caspase-3 experssion was decaeased Naloxone at 10 μmol·L-1 inhibited the promoting effects of morphine;GF109203X at 10 μmol·L-1 or Staurosporine at 1 μmol·L-1 could inhibit the promoting effects of morphine,manifestation,apoptosis rates were increased,Caspase-3 experssion was incaeased,PKC expression was decreased.Conclusion morphine inhibited serum hungry-induced apoptosis in cardiac myocytes via PKC signal transduction pathway.