【作者】 祖秀宏；

【导师】 王宗保； 易光辉；

【作者基本信息】 南华大学 ， 病理学与病理生理学， 2007， 硕士

【摘要】 目的:观察氧化低密度脂蛋白(ox-LDL)和高糖对人脐静脉内皮细胞-12细胞株(HUVEC-12) LOX-1和IL-1β表达的影响,以及PPARγ激动剂ciglitazone对ox-LDL和高糖诱导的LOX-1表达的影响。方法:分别用不同浓度的ox-LDL(0、20、40、80μg/ml)和不同浓度的葡萄糖(0、5.6、11.2、22.4、44.8mmol/L及甘露醇对照组)与HUVEC-12共同孵育24h,然后将ox-LDL和22.4mmol/L葡萄糖与HUVEC-12共孵育24小时,用RT-PCR、流式细胞仪分别检测细胞中LOX-1 mRNA和蛋白质的表达;用RT-PCR检测细胞IL-1βmRNA的表达,酶联免疫吸附实验(ELISA法)检测细胞培养液中IL-1β的分泌量。分别用40μg/ml的ox-LDL和22.4mmol/L高糖加入10μmol/L的PPARγ激动剂ciglitazone共孵育HUVEC-12细胞24小时,用RT-PCR和Western blot分别检测细胞中LOX-1 mRNA和蛋白质的表达。结果:随着ox-LDL和D-葡萄糖浓度的增加,LOX-1 mRNA和蛋白质的表达逐渐增加,IL-1βmRNA的表达和分泌量也逐渐增加。用40μg/ml的ox-LDL和22.4mmol/L的高糖共孵育HUVEC-12细胞24小时后,LOX-1和IL-1β的表达也是上调的。用10 ng/ml的IL-1β处理HUVEC-12细胞24小时后,与空白对照组相比,LOX-1 mRNA和蛋白质的表达也是增加的。分别用40μg/ml的ox-LDL和22.4mmol/L高糖加入10μmol/L的ciglitazone共同孵育HUVEC-12细胞24小时后,同空白对照组相比, ciglitazone可以降低ox-LDL和高糖诱导的LOX-1的表达。结论: Ox-LDL和高糖呈剂量依赖性增加人脐静脉内皮细胞LOX-1和IL-1β的表达水平;IL-1β也可上调人脐静脉内皮细胞LOX-1的表达;PPARγ激动剂ciglitazone可以降低ox-LDL和高糖诱导的LOX-1的表达。更多还原

【Abstract】 OBJECTIVE To investigate the effects of oxidized low density lipoprotein(ox-LDL) and/or high glucose on expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) and interleukin-1β(IL-1β) in human umbilical vein endothelial cell line (HUVEC-12), and the inhibitory effect of PPARγagonist on ox-LDL and/or high glucose- induced expression of LOX-1.METHODS Human umbilical vein endothelial cells-12 were treated with ox-LDL at different concentrations (0,20,40,80μg/ml) and glucose (0,5.6, 11.2, 22.4, 44.8mmol/L) for 24h, respectively. LOX-1 mRNA expression levels were determined by reverse transcription-polymerase chain reaction (RT-PCR),and the protein expression levels were evaluated by Western blot and flow cytometry, respectively. Interleukin-1βmRNA and protein levels were determined by reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells-12 were co-incubated with ox-LDL (40μg/ml), high glucose(22.4 mmol/L), or combination of oxLDL(40μg/ml) and high glucose (22.4 mmol/L) for 24h. Furthermore, the cells as described above were treated with PPARγagonist ciglitazone (10μmol/L) to illustrate the inhibitory effect of PPARγagonist on the induced expression of LOX-1 by ox-LDL or high glucose.RESULTS The results showed that ox-LDL (20, 40, 80μg/ml) and high glucose (22.4, 44.8 mmol/L) increased LOX-1and IL-1βexpression in both protein and mRNA levels in a dose-dependent manner, respectively. LOX-1 expression was also up-regulated by IL-1βas showed in both protein and mRNA levels. PPARγagonist ciglitazone inhibited the upregulation of LOX-1 expression induced by ox-LDL or high glucose as in both protein and mRNA levels. CONCLUSION Ox-LDL and high glucose up-regulate the expression of LOX-1 and IL-1βin a concentration-dependent manner. IL-1βmay up-regulate LOX-1 expression. PPARγagonist ciglitazone may decrease LOX-1 expression level induced by ox-LDL or high glucose.更多还原