氯化锂调节趋化因子Fractalkine表达抑制缺氧复氧诱导的血管内皮细胞损伤

The cytoprotective effects of Lithium chloride on HUVECs through regulating fractalkine during the anoxia/reoxygenation

目的:观察缺氧复氧诱导人脐静脉血管内皮细胞(Human umbillcal vein endothelial cells,HUVECs)损伤和趋化因子(Fractalkine,FKN)表达变化及氯化锂(Lithium chloride,LiCl)的干预效应。方法:体外培养HUVECs并随机分为对照组、缺氧复氧组和LiCl 5、10 mmol/L组。MTT法测定HUVECs的存活率,流式细胞术检测HUVECs凋亡率,免疫细胞荧光技术检测HUVECs的FKN蛋白表达,RT-PCR技术检测HUVECs的FKN mRNA表达。结果:与对照组比较,缺氧复氧组HUVECs存活率显著降低但凋亡率显著增高(P<0.01)且FKN mRNA和蛋白质表达也显著增高(P<0.01);与缺氧复氧组比较,LiCl 5、10 mmol/L组HUVECs存活率显著增高但凋亡率显著降低(P<0.01)且FKNmRNA和蛋白质表达也明显降低(P<0.05)。结论:缺氧复氧上调FKN表达诱导HUVECs损伤(存活率降低、凋亡率增加);LiCl预处理能够下调FKN表达,显著减轻缺氧复氧导致的HUVECs损伤。

Objective：To investigate the cytoprotective effects of lithium chloride（LiCl） on human umbillcal vein endothelial cells（HUVECs） through regulating Fractalkine（FKN） during the anoxia/reoxygenation.Methods：HUVECs were cultivated and divided into four groups randomly（control group,anoxia/reoxygenation group,LiCl 5 mmol/L group,and LiCl 10 mmol/L group）,which were used to establish the anoxia/reoxygenation models.The cell viability was detected by MTT method.The apoptosis rate of HUVECs was examined by Flow cytometry.The mRNA expression of FKN was assessed by RT-PCR.The intracellular distribution and expression of FKN were determined by immunofluorescent staining.Results：Compared with control group,the apoptotic rate of HUVECs increased but their viability rate reduced significantly in anoxia/reoxygenation group（all P0.01）.The mRNA and protein expression of FKN were increased remarkably in anoxia/reoxygenation group compared to control group（P0.01）.Compared with anoxia/reoxygenation group,the survival rate of HUVECs was increased but their apoptotic rate as well as the expression of FKN mRNA and FKN protein reduced significantly in LiCl 5 mmol/L and 10 mmol/L groups（P0.01 or P0.05）.Conclusions：The anoxia/reoxygenation induced the damage of HUVECs through upregulating FKN expression.LiCl pretreatment can protect HUVECs against anoxia/reoxygenation injury via inhibiting FKN expression.