摘要
目的:蛋白激酶C(PKC)活化对L-6TG大鼠肌母细胞缺血/再灌注损伤过程中细胞凋亡的影响。方法:将培养的L-6TG大鼠肌母细胞随机分为3组:①正常对照组(C组);②缺血/再灌注组(I/R组);③PMA+缺血/再灌注组(PMA组)。观测了细胞内SOD、XOD、Ca2+含量的变化;采用MTT法检测线粒体的功能;利用流式细胞仪和细胞DNA电泳结果检测细胞凋亡情况;采用免疫组织化学的方法检测caspase-3的蛋白表达情况,结合自动图像分析系统对其结果进行定量分析。结果:蛋白激酶C活化可显著降低L-6TG大鼠肌母细胞I/R 4 h后细胞内XOD、Ca2+含量及凋亡细胞百分率,增加细胞内SOD活性及线粒体呼吸功能,DNA电泳无梯状条带出现,caspase-3的表达明显下调。结论:蛋白激酶C活化可明显减轻L-6TG大鼠肌母细胞缺血再灌注损伤后的细胞凋亡的发生,其机制可能与减轻氧化损伤、调节细胞内钙稳态、减轻线粒体损伤、减少caspase-3表达有关。
Aim： To study the effects of PKC activition on apoptosis during ischemia/reperfusion in L-6TG rat skeletal myoblasts. Me thods： Cultured L-6TG cells were divided into 3 groups： control group （C）, ischemia/reperfusion group（I/R）, PMA ＋ ischemia/ reperfusion group（PMA）. SOD, XOD and free calcium and mltochondral respiration in L-6TG cell were evaluated in each group. Apoptosis was detected by flow cytometer with PI staining method and agarose gel electrophoresis, the immunohistochemical method was used to determine the expression of caspase-3. Results： Compared with I/R group, in PMA group, XOD , free calcium in L- 6TG cell and apoptotic percentage all decreased significantly, while SOD and mitochondral respiration in L-6TG ceil increased. DNA fragmentation analysis of L 6TG cell showed no laddering pattern. The expression of caspase-3 was down regulated significantly. Conclusion： Activation of PKC can lessen ischemla/reperfusion injury and apoptosis through lessening oxidative injury and mitochondrial inury, adjusting calcium dyshomeostasis and down expression of caspase-3.
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2005年第4期437-440,共4页
Chinese Journal of Applied Physiology
