lncRNA DINO在同型半胱氨酸致内皮细胞凋亡中的作用

The Study of the Role of LncRNA DINO in Endothelial Cell Apoptosis Induced by Homocysteine

目的探讨DNA损伤诱导的长链非编码RNA(long noncoding RNA,lncRNA)DINO在同型半胱氨酸(homocysteine,Hcy)致内皮细胞凋亡中的作用。方法以人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)为研究对象,MTT法检测不同浓度的Hcy对内皮细胞增殖的影响,筛选合适浓度;最适浓度Hcy干预细胞后分为对照组(0μmol·L^(-1) Hcy)和实验组(100μmol·L^(-1) Hcy),用细胞活力染色及流式细胞术检测细胞凋亡情况,实时荧光定量PCR(RT-qPCR)检测lncRNA DINO的表达;细胞活力染色、流式细胞术、蛋白免疫印迹(Western blot)分别检测Hcy作用于转染lncRNA DINO干扰片段的HUVECs细胞的凋亡情况及凋亡相关蛋白Bax、Bcl-2的表达。结果100μmol·L^(-1) Hcy对内皮细胞的增殖有抑制作用(P<0.001);与对照组相比,实验组细胞凋亡数增加,且lncRNA DINO表达增加(P均<0.01);转染干扰片段si-DINO、lncRNA DINO表达下降(P<0.01);Hcy作用于转染lncRNA DINO干扰片段的HUVECs细胞的凋亡数下降,Bax/Bcl-2比值下降(P均<0.01)。结论lncRNA DINO能够促进Hcy诱导的内皮细胞凋亡。

Objective To investigate the role of non-coding RNA(lncRNA)DINOinduced by DNA damage in endothelial cell apoptosis induced by homocysteine(Hcy).Methods Human umbilical vein endothelial cells were used,and the effects of different concentrations of Hcy on endothelial cell proliferation were detected by MTT method,and the appropriate concentration was screened.After treated with the optimal concentration of Hcy,the cells were divided into control group(0μmol·L^(-1) Hcy)and experimental group(100μmol·L^(-1) Hcy).Apoptosis was detected by cell viability staining and flow cytometry,and the expression of lncRNA DINO was detected by qPCR.After transfection of lncRNA DINO interference fragment,apoptosis was detected by cell viability staining,flow cytometry and the expression of apoptosis-related proteins Bax and Bcl-2 were detected by Western blot,respectively.Results The results showed that Hcy had a significant inhibitory effect on endothelial cells at 100μmol·L^(-1)(P<0.001).Compared with the control group,the number of apoptosis and the expression of lncRNA DINO were significantly increased in the experimental group(P all<0.01),and the expression of lncRNA DINO was decreased by transfection interference fragment si-DINO(P all<0.01).The apoptosis number of HUVECs cells decreased after transfection of interference fragments and Hcy intervention,and the ratio of Bax/Bcl-2 was decreased(P all<0.01).Conclusion lncRNA DINO can promote endothelial cell apoptosis induced by Hcy.