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目的:观察注射用血栓通(主要成分为三七总皂苷)对过氧化氢与肿瘤坏死因子(TNF-α)诱导大鼠血管内皮细胞凋亡的保护效应,并对其干预血管内皮细胞的凋亡通路和机制进行研究,为注射用血栓通更广泛的临床应用提供试验依据。方法:针对体外培养的大鼠肺部血管内皮细胞,施加过氧化氢与TNF-α等凋亡诱导因子,建立血管内皮细胞凋亡诱导模型,继而采用50、100μg/mL等浓度的注射用血栓通进行预干预,并采用Annexin V Ca2+依赖性磷脂结合蛋白/PI碘化丙啶染色法检测血管内皮细胞凋亡率以及采用细胞线粒体膜电位变化反映细胞线粒体损伤率,进而检测凋亡相关通路调控基因Caspase-3、Bcl-2和Fas的表达情况。结果:注射用血栓通浓度为100μg/mL时,能使过氧化氢诱导的大鼠血管内皮细胞凋亡率与线粒体损伤率显著降低,但对TNF-α诱导的细胞凋亡无显著的抑制作用,进一步研究发现以100μg/mL的注射用血栓通可明显减低过氧化氢诱导的Caspase-3表达,而且还可明显提高Bcl-2的水平,但对Fas表达的影响不明显。结论:注射用血栓通(三七总皂苷)可通过下调Caspase-3表达与上调Bcl-2基因来抑制氧化应激诱导的血管内皮细胞凋亡。
OBJECTIVE: To observe the protective effect of Xuesaitong injection (the main component of Panax notoginseng saponins) on the apoptosis of vascular endothelial cells induced by hydrogen peroxide and tumor necrosis factor (TNF-α), and to observe its effect on the apoptosis of vascular endothelial cells Death pathways and mechanism of the study for the more general clinical application of thrombosis injection provides the experimental basis. Methods: Apoptosis inducing model of vascular endothelial cells was established by applying H2O2 and TNF-α and other apoptosis-inducing factors to rat pulmonary vascular endothelial cells cultured in vitro, followed by 50, 100μg / mL injection of thrombus Apoptotic rate of vascular endothelial cells was detected by Annexin V Ca2 + -dependent phospholipid binding protein / PI propidium iodide staining and the change of mitochondrial membrane potential was used to detect the mitochondrial damage rate, then the apoptosis-related pathway was detected Gene Caspase-3, Bcl-2 and Fas expression. Results: When Xueshuantong injection was 100μg / mL, hydrogen peroxide could significantly decrease the apoptosis rate and mitochondrial damage rate of vascular endothelial cells, but did not inhibit TNF-α-induced apoptosis significantly Further study found that 100μg / mL injection of thrombus can significantly reduce hydrogen peroxide-induced Caspase-3 expression, but also significantly increased Bcl-2 levels, but the impact of Fas expression was not obvious. Conclusion: Xueshuantong Injection (Panax notoginseng saponins) can inhibit oxidative stress-induced apoptosis of vascular endothelial cells by down-regulating Caspase-3 expression and up-regulating Bcl-2 gene.