摘要
目的建立氧化低密度脂蛋白(ox—LDL)诱导培养人脐静脉内皮细胞(HUVECs)凋亡模型,探讨促红细胞生成素(EPO)对ox—LDL诱导HUVECs凋亡的影响。方法取体外培养3~6代的HUVECs用于实验。实验分为两组:一组细胞予以不同浓度(6.25、50、100kU/L)重组人促红细胞生成素(rhEPO)预处理24h,再加入100mg/LOX—LDL孵育48h;另一组细胞加入反式或顺式LOX-1 mRNA预处理24h,再加入100mg/L的ox—LDL孵育12h。采用存活率、凋亡率和Bcl-2/Bax比值评价细胞凋亡情况。结果与ox—LDL组比较,随rhEPO浓度的递增,细胞存活率增高,细胞凋亡率降低,凋亡蛋白Bcl-2/Bax比值增高(P均〈0.05),呈剂量依赖性。反式LOX-1 mRNA(0.5μmol/L)预处理组凋亡蛋白Bcl-2/Bax比值较OX—LDL组明显增高(P〈0.05);顺式LOX-1 mRNA(0.5μmol/L)预处理组Bcl-2/Bax比值与ox—LDL组则无明显差异。结论ox—LDL可以诱导HUVECs凋亡,并且通过LOX-1 mRNA来调节,rhEPO可增高Bcl-2/Bax比值,抑制ox—LDL诱导的HUVECs凋亡。
Objective To explore the protective effect of erythropoietin (EPO) against oxidized-low density lipoprotein (ox-LDL)-induced apoptosis in human umbilical vein endothelial ceils (HUVECs) in an ox-LDL induced apoptosis model. Methods Third - sixth passage of cultured HUVECs were used, and they were divided into two groups. The cells were pretreated with different concentrations (6.25, 50, 100 kU/L) of recombinant human erythropoietin (rhEPO) for 24 hours, then they were exposed to ox-LDL (100 mg/L) for 48 hours; another group of cells were pretreated with antisense to 0.5 μmol/L LOX-1 mRNA or 0. 5 μmol/L sense for 24 hours, and then HUVECs were exposed to ox-LDL (100 mg/L) for 12 hours. Apoptosis was assessed by the apoptosis ratio, cell viability, and Bcl-2/Bax ratio. Results As compared to untreated controls, pretreatment with rhEPO led to increased cell survival of HUVECs and decreased cell apoptosis in a dose-dependent manner (all P〈0.05). Consistently, the Bcl-2/Bax ratios were also increased in a similar fashion. The ratio of apoptosis protein Bcl-2/Bax was increased in the antisense LOX-1 mRNA group than that of ox-LDL group (P〈0.05), but the one in the sense LOX-1 mRNA group was not significantly different from that of ox-LDL group. Conclusion The ox-LDL can induce apoptosis in HUVECs by regulating LOX-1 mRNA, and rhEPO can increase Bcl-2/Bax ratio and inhibite ox-LDL-induced apoptosis of HUVECs.
出处
《中国危重病急救医学》
CAS
CSCD
北大核心
2009年第11期656-659,共4页
Chinese Critical Care Medicine
基金
甘肃省新药临床前研究重点实验室开放基金项目(GSKFKT-0707)
兰州大学医学科研基金项目(820726)