Ghrelin抑制高糖诱导的血管内皮损伤的作用及其机制

Inhibitory effect of ghrelin on vascular endothelial injury induced by high glucose and its mechanism

目的探究Ghrelin对高糖诱导的内皮损伤的保护作用及机制。方法将人血管内皮细胞(HVECs)分为5mmol/L d-葡萄糖组、30mmol/L d-葡萄糖组、30mmol/L d-葡萄糖+4mmol/L ghrelin组、30mmol/L d-葡萄糖+20mmol/L ghrelin组和30mmol/L d-葡萄糖+100mmol/L ghrelin组。采用流式细胞术检测细胞凋亡,采用MTT法检测细胞活性,采用过氧化物敏感性荧光探针二氯二氢荧光素二乙酸酯(DCFH-DA)检测细胞内ROS水平,采用Western blot检测细胞胞浆中p65、HO-1、Bcl-2、Bax和cleaved caspase 9水平。结果与5mmol/L d-葡萄糖组比较,30mmol/L d-葡萄糖组细胞凋亡明显增加,活性降低,ROS生成增加和cytosolic p65和Bax水平均显著降低,HO-1、Bcl-2和cleaved caspase 9水平显著升高;与30mmol/L d-葡萄糖组比较,ghrelin治疗组的细胞活性明显增加,凋亡率明显降低,ROS产生减少,胞浆p65和Bax水平均显著升高,HO-1、Bcl-2和cleaved caspase 9水平均显著降低。结论ghrelin可抑制高糖诱导的血管内皮损伤,其机制与抗ROS生成、抑制HO-1和NF-κB信号活性相关。

Objective To investigate the protective effect of ghrelin on high glucose-induced endothelial injury and its mechanism.Methods Human vascular endothelial cells(HVECs)were divided into groups as 5 mmol/L d-glucose group,30 mmol/L d-glucose group,30 mmol/L d-glucose+4 mmol/L ghrelin group,30 mmol/L d-glucose+20 mmol/L ghrelin group and 30 mmol/L d-glucose+100 mmol/L ghrelin group.The apoptosis of each group was detected by flow cytometry.The cell viability of each group was detected by MTT assay.The intracellular ROS levels of each group were detected by peroxide sensitive fluorescent probe dichlorodihydrofluorescein diacetate(DCFH-DA).The expression levels of cytosolic p65,HO-1,Bcl-2,Bax and cleaved caspase 9 proteins were detected by Western blot.Results Compared with the 5 mmol/L d-glucose group,the 30 mmol/L d-glucose group showed significantly increased apoptosis rate and decreased cell viability,increased ROS production,but significantly decreased protein expression levels of cytosolic p65 and Bax,and significantly increased expression levels of HO-1,Bcl-2 and cleaved caspase 9.Compared with the 30 mmol/L d-glucose group,ghrelin treatment groups showed significantly increased cell viability,decreased apoptosis rate,decreased ROS production,significantly increased protein expression levels of cytosolic p65 and Bax,and significantly decreased expression levels of HO-1,Bcl-2 and cleaved caspase 9.Conclusion Ghrelin can inhibit endothelial injury induced by high glucose,and its mechanism is related to anti-ROS production and inhibition of HO-1 and NF-κB signaling activity.