山姜素下调微小RNA146a-5p表达减轻血管内皮细胞氧化应激损伤

Alpinetindown regulates miR146a-5p expression and alleviates the endothelium oxidative stress injury induced by endotoxin

目的:探讨山姜素调控微小RNA(miR)146a-5p表达对内毒素诱导的血管内皮细胞氧化应激损伤的影响。方法:采用1μg/mL的脂多糖处理人脐静脉血管内皮细胞(HUVEC)构建细胞损伤模型。将HUVEC分为对照组、LPS组、LPS+山姜素低、中、高剂量组、LPS+anti-miR阴性对照组、LPS+anti-miR-146a-5p组、LPS+山姜素+miR-NC阴性对照组、LPS+山姜素+miR-146a-5p组。采用生化法检测超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性以及丙二醛(MDA)水平。采用流式细胞术检测HUVEC的凋亡率。采用实时定量聚合酶链式反应(RT-qPCR)检测miR-146a-5p表达。结果:与Con组比较,LPS组细胞凋亡率、MDA水平、miR-146a-5p表达显著升高,SOD和GSH-Px活性显著降低。与LPS组比较,LPS+山姜素低、中、高剂量组细胞凋亡率、MDA水平、miR-146a-5p表达显著降低,而SOD和GSH-Px活性显著升高(P均<0.05)。与LPS+anti-miR-阴性对照组比较,LPS+anti-miR-146a-5p组细胞凋亡率、MDA水平显著降低,而SOD和GSH-Px活性显著升高(P均<0.05)。与LPS+山姜素+miR-阴性对照组比较,LPS+山姜素+miR-146a-5p组细胞凋亡率、MDA水平显著升高,而SOD和GSH-Px活性显著降低(P均<0.05)。结论:山姜素通过下调miR-146a-5p表达减轻内毒素诱导的血管内皮细胞氧化应激损伤。

Objective:To investigate the effect of alpinetin on endotoxin-induced oxidative stress injury in vascular endothelial cells by regulating miR146a5p expression.Methods:Human umbilical vein endothelial cells(HUVECs)were treated with 1μg/mL lipopolysaccharide(LPS)to construct a cell injury model.HUVECs were divided into control(Con)group,LPS group,LPS+alpinetin(low dose)group,LPS+alpinetin(medium dose)group,LPS+alpinetin(high dose)group,LPS+antimiRNC group,LPS+antimiR146a5p group,LPS+alpinetin+miRNC group,LPS+alpinetin+miR146a5p group.Biochemical methods were applied to detect the activities of superoxide dismutase(SOD)and glutathione peroxidase(GSHPx),and the level of malondialdehyde(MDA).Flow cytometry was used to detect the apoptosis rate of HUVECs.Real-time quantitative PCR(RTqPCR)was employed to detect the miR146a5p expression.Results:As compared with the Con group,the apoptosis rate,MDA level,and miR146a5p expression in the LPS group were significantly increased(P<0.05),and the SOD and GSHPx activities were significantly decreased(P<0.05).As compared with the LPS group,the apoptosis rate,MDA level and miR146a5p expression in the LPS+alpinetin(low dose)group,LPS+alpinetin(medium dose)group and LPS+alpinetin(high dose)group were significantly reduced(P<0.05),SOD and GSHPx activities were significantly increased(P<0.05).As compared with the LPS+antimiRNC group,the apoptosis rate and MDA level in the LPS+antimiR146a5p group were significantly reduced(P<0.05),and the activities of SOD and GSHPx were significantly increased(P<0.05).As compared with the LPS+alpinetin+miRNC group,the apoptosis rate and MDA level in the LPS+alpinetin+miR146a5p group were significantly increased(P<0.05),and SOD and GSHPx activities were significantly reduced(P<0.05).Conclusion:Alpinetin could reduce endotoxin-induced oxidative stress damage to vascular endothelial cells by down-regulating the miR146a5p expression.