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目的:探明白花丹参对脂多糖处理的血管内皮细胞凋亡的影响及作用机制。方法:内毒素处理培养的人脐静脉内皮细胞造成细胞凋亡。培养液中添加系列浓度的白花丹参水溶性成分培养细胞,倒置相差显微镜观测细胞形态变化。吖啶橙染色,共聚焦显微镜观测凋亡细胞核片段化状况;PI染色,流式细胞仪检测细胞周期分布。生化试剂盒检测细胞内脂质过氧化物(MDA)含量,超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)活性。结果:脂多糖处理使血管内皮细胞形成大量凋亡小体,细胞核片段化严重,并明显降低了细胞存活率,改变了细胞周期的正常分布,阻止细胞由G1期进入S期,同时显著上调了细胞中的MDA含量,而下调了其中的SOD、GSH-PX活性明显。白花丹参水溶性部位在一定浓度范围内,剂量依赖性地逆转脂多糖引起的上述变化。结论:白花丹参水溶性部位可能通过调节细胞氧化应激状态抑制脂多糖导致的血管内皮细胞凋亡。
Objective: To investigate the effect of Salvia miltiorrhiza on the apoptosis of vascular endothelial cells treated with lipopolysaccharide and its mechanism. Methods: Endotoxin treatment of cultured human umbilical vein endothelial cells caused apoptosis. A series of water-soluble components of Radix Salviae Miltiorrhizae were added to the culture solution, and the morphological changes of the cells were observed by inverted phase contrast microscope. Acridine orange staining, confocal microscopy of apoptotic nucleus fragmentation status; PI staining, flow cytometry cell cycle distribution. The biochemical kit was used to detect the contents of intracellular lipid peroxide (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX). Results: LPS treatment resulted in the formation of a large number of apoptotic bodies in endothelial cells, severely fragmented nuclei and markedly decreased cell viability, altered the normal distribution of cell cycle, prevented the cells from entering the S phase from G1 phase and significantly upregulated Cell MDA content, which down-regulated SOD, GSH-PX activity significantly. Water-soluble Salvia miltiorrhiza parts in a certain concentration range, dose-dependent reversal of lipopolysaccharide-induced changes. Conclusion: The water-soluble fraction of Salvia miltiorrhiza may inhibit the apoptosis of vascular endothelial cells induced by lipopolysaccharide by regulating the cellular oxidative stress.