蒺藜皂苷激活PKCε抗氧化应激诱导心肌细胞凋亡的机制
Mechanisms of gross saponins of Tribulus terrestris via activating PKCε against myocardial apoptosis induced by oxidative stress
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摘要:
观察蒺藜皂苷 (gross saponins of Tribulus terrestris, GSTT) 在过氧化氢 (hydrogen peroxide, H2O2) 诱导新生大鼠心肌细胞凋亡过程中对蛋白激酶C Eplison亚型 (protein kinase C Eplison, PKCε) 及凋亡相关蛋白的影响, 探讨蒺藜皂苷抗心肌细胞凋亡的作用机制。新生大鼠心肌细胞原代培养, H2O2建立心肌细胞凋亡模型, 流式细胞术检测细胞凋亡率; Western blotting法检测phospho-PKCε及Bcl-2、Bax的蛋白含量; 免疫细胞化学技术检测cleaved caspase-3蛋白含量。与模型组比较, GSTT (100 mg·L-1) 预处理可明显降低H2O2诱导的心肌细胞凋亡率 (P < 0.01), 并增加phospho-PKCε及Bcl-2的蛋白含量 (P < 0.01), 降低Bax及cleaved caspase-3的蛋白含量 (P < 0.01); PKC抑制剂白屈菜红碱 (chelerythrine, Che) 可部分阻断蒺藜皂苷抗心肌细胞凋亡作用, 与蒺藜皂苷组比较具有统计学意义 (P < 0.05, P < 0.01)。GSTT抗心肌细胞凋亡的作用机制可能与激活PKCε后抑制线粒体依赖的凋亡有关。
Abstract:This study is to observe the effect of gross saponins of Tribulus terrestris (GSTT) on protein kinase Cε (PKCε) and apoptosis-associated protein in the apoptosis of cultured cardiocyte apoptosis induced by hydrogen peroxide (H2O2), and to explore the mechanisms of GSTT against myocardial apoptosis. Primary cardiocytes were isolated and cultured. Myocardial apoptosis was induced by H2O2 and analyzed with flow cytometry. Protein content of phospho-PKCε, Bcl-2, and Bax were detected with Western blotting analysis. Cleaved caspase-3 protein content was determined with immunocytochemical technique. After the pretreatment of 100 mg·L-1 GSTT, compared with H2O2 group, GSTT could not only decrease the apoptotic percentage in cardiocytes damaged by H2O2 (P < 0.01), but also reduce protein contents of Bax and cleaved caspase-3 (P < 0.01), and increase protein content of phospho-PKCε and Bcl-2 significantly (P < 0.01). PKC inhibitor chelerythrine (Che) could prevent partly the effect of GSTT against myocardial apoptosis (P < 0.05 and P < 0.01). Mechanisms of GSTT against myocardial apoptosis might be associated with inhibition of mitochondrial apoptosis pathway after PKCε activation.
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