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目的:研究ghrelin对ox-LDL诱导的人脐静脉内皮细胞(HUVEC)凋亡的抑制作用。方法:采用Ho-echst 33258荧光染色、Western blot法和硝酸还原酶法研究ghrelin对ox-LDL诱导的人脐静脉内皮细胞凋亡的抑制作用。结果:Hoechst 33258染色荧光检测发现,与正常对照组相比,ox-LDL组HUVEC中出现大量凋亡细胞,凋亡率明显增加,与ox-LDL组比较,ghrelin预处理组HUVEC细胞凋亡率呈剂量依赖性下降;Western blot检测结果表明,与对照组比较,ox-LDL可上调内细胞中Caspase-3表达,而经过ghrelin预处理后能剂量依赖性降低Caspase-3蛋白表达;硝酸还原酶法结果显示,ox-LDL可显著减少内皮细胞培养液中NO含量,经过ghrelin预处理后,ghrelin能剂量依赖性增加NO含量。结论:ghrelin对ox-LDL诱导的人脐静脉内皮细胞凋亡有拮抗作用,可能与其抑制ox-LDL引起的细胞内Caspase-3蛋白水平升高及NO合成下降有关。
AIM: To investigate the inhibitory effect of ghrelin on the apoptosis of human umbilical vein endothelial cells (HUVECs) induced by ox-LDL. Methods: The inhibitory effect of ghrelin on the apoptosis of human umbilical vein endothelial cells induced by ox-LDL was studied by Ho-echst 33258 staining, Western blot and nitrate reductase method. Results: Hoechst 33258 staining showed that compared with the normal control group, a large number of apoptotic cells were found in the HUVECs in the ox-LDL group and the apoptosis rate was significantly increased. Compared with the ox-LDL group, the apoptosis rate of HUVECs in ghrelin preconditioning group The results of Western blot showed that compared with the control group, ox-LDL up-regulated the expression of Caspase-3 in the cells, while the ghrelin pretreatment reduced the expression of Caspase-3 protein in a dose-dependent manner. The activity of nitrate reductase The results showed that ox-LDL could significantly reduce the content of NO in endothelial cell culture medium, ghrelin could increase NO content in a dose-dependent manner after ghrelin pretreatment. CONCLUSION: ghrelin antagonizes ox-LDL-induced apoptosis in human umbilical vein endothelial cells and may be related to the inhibition of ox-LDL-induced increase of intracellular levels of Caspase-3 protein and decrease of NO synthesis.