神经酰胺上调bax表达诱导人内皮细胞凋亡  

Ceramide induces apoptosis of human endothelium via up-regulation of bax expression

作　　者：沈珠甫[1] 余德文[1] 陈代陆[1] 杨宇[1] 谭志胜[1] 王湘平[1] 梁源[1] 刘丹[1] 

机构地区：[1]成都军区昆明总医院干部病房,650032

出　　处：《西南国防医药》2010年第2期117-119,共3页Medical Journal of National Defending Forces in Southwest China

基　　金：云南省自然科学基金资助项目(2006C0059M)

摘　　要：目的观察外源性神经酰胺(C2-ceramide)对内皮细胞凋亡及相关基因bax表达变化的影响。方法原代培养人脐静脉血管内皮细胞,用不同浓度的ceramide干预内皮细胞24h,并用同一浓度的ceramide作用内皮细胞不同时间后,采用TUNEL法检测细胞凋亡,逆转录聚合酶链反应(RT-PCR)、Western blot技术对bax mRNA及蛋白表达分析。结果神经酰胺处理组内皮细胞的凋亡阳性率显著高于对照组(P<0.05),且具有时间和剂量依赖性;对照组和5、12.5、25、50μmol/L神经酰胺处理24h后,各组细胞凋亡率随干预剂量的加大逐渐上升;终浓度25μmol/L的神经酰胺处理6、24、48h后,细胞凋亡率随时间延长而逐渐升高;神经酰胺处理组bax mRNA及蛋白的表达较对照组显著升高(P<0.01～0.05)。结论外源性神经酰胺能通过上调bax表达诱导内皮细胞凋亡。Objective To observe the effects of exogenous ceramide on apoptosis of endothelial cells（EC） and the expression of related bax gene. Methods The vascular endothelial cells of human umbilical vein were primarily cultured, and then they were exposed to different doses of ceramide for 24 h and the same dose of ceramide for different times. The apoptosis indexes were determined by TUNEL method, and bax mRNA and bax protein expression of EC were detected by RT - PCR and Western blot technique, respectively. Results The number of apoptotic cells was significantly higher in ceramide - treated EC than that in control group （P 〈 0.05 ）. In addition, ceramide - induced EC apoptosis was in time - and dose - dependent manner. After ceramide treatment for 24 h, the percentage of the apoptotic EC increased gradually with the increase of ceramide dosage. After 25 μmol/L of ceramide treatment for 6,24 and 48 h, the rate of apoptotic cells increased with the time passing. The expressions of bax mRNA and protein in EC treated with ceramide significantly increased compared with control group （ P 〈 0.01 - 0.05 ）. Conclusion Ceramide may induce apoptosis of endothelial cells via up - regulating the bax mRNA and protein expression.

关 键 词：神经酰胺 内皮细胞 凋亡 BAX 

分 类 号：Q25[生物学—细胞生物学]