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目的:研究西红花苷对培养的牛血管内皮细胞凋亡的影响。方法:采用3β、5α、6β三羟胆固(烷)醇(Triol)诱导内皮细胞凋亡的模型,用比色法测定丙二醛含量,光学显微镜、电子显微镜检测形态和超微结构的变化,DNA电泳检测DNAladder及流式细胞仪分析法检测凋亡率,采用逆转录聚合酶链式反应(RTPCR)法检测BaxmRNA表达的变化。结果:Triol处理后,培养液中MDA含量增加,为1.761nmol·L-1(P<0.01),细胞收缩,核浓缩,深染,线粒体肿胀空化,出现凋亡小体,出现凋亡典型的“DNAladder”,和亚二倍体峰,凋亡率为30.62%,BaxmRNA表达量增加;西红花苷(10-7,10-6mol·L-1)+Triol组,MDA含量减少,内皮细胞形态结构的完整,凋亡率分别为24.4%,6.3%,BaxmRNA表达量降低。结论:西红花苷可能是抗脂质过氧化并通过调节BaxmRNA表达,减少细胞异常凋亡。
Objective: To study the effect of crocin on the apoptosis of cultured bovine vascular endothelial cells. METHODS: Apoptosis model of endothelial cells induced by 3β, 5α and 6β Triol was measured. The content of malondialdehyde was measured by colorimetry. The changes of morphology and ultrastructure were detected by light microscope and electron microscope. Apoptosis rate was detected by DNA ladder and flow cytometry analysis. The expression of Bax mRNA was detected by reverse transcription polymerase chain reaction (RTPCR). RESULTS: After Triol treatment, the content of MDA in the culture fluid increased to 1.761 nmol·L-1 (P<0.01). The cells contracted, the nucleus was concentrated and deep stained, and the mitochondria swelled with cavitation, apoptotic bodies appeared, and typical apoptosis appeared. The “DNA ladder”, and the hypodiploid peak, the apoptotic rate was 30.62%, and the expression of Bax mRNA was increased. The content of MDA was decreased in the crocin group (10-7, 10-6 mol.L-1) + Triol. The integrity of the morphological structure of the cells, the apoptotic rates were 24.4%, 6.3%, and the expression of Bax mRNA was decreased. Conclusion: Crocin may be anti-lipid peroxidation and reduce Bax mRNA expression and reduce abnormal apoptosis.