人剪切修复基因D介导ox-LDL促进HUVEC凋亡作用

目的探讨人剪切修复基因D(XPD)是否介导氧化型低密度脂蛋白(ox-LDL)促进人脐静脉内皮细胞(HUVEC)凋亡。方法以ox-LDL建立HUVEC凋亡模型。用脂质体将XPD-siRNA转染HUVEC,给予ox-LDL处理。实验分6组:空白对照组;阴性对照siRNA组;XPD-siRNA组;ox-LDL组;ox-LDL+阴性对照siRNA组;ox-LDL+XPDsiRNA组。MTT测定细胞活力;流式细胞仪检测细胞凋亡率和细胞周期;用RT-PCR和Western blot检测XPD、Bax和Bcl-2的表达。结果建立HUVEC凋亡模型ox-LDL的最佳浓度为100 mg/L。与阴性对照siRNA组相比,XPDsiRNA组的细胞凋亡率明显下降(P0.05),存活率明显增加(P0.01),G0/G1期细胞减少(P0.05)、S期细胞增加(P0.05),XPD、Bax表达降低(P均0.05),Bcl-2表达增高(P0.05);与空白对照组相比,ox-LDL组细胞凋亡率明显增加(P0.01),细胞存活率下降(P0.05),G0/G1期细胞增加(P0.05)、S期细胞明显减少(P0.01),XPD、Bax表达升高(P均0.05),Bcl-2表达降低(P0.05);与ox-LDL+阴性对照siRNA组相比,ox-LDL+XPD-siRNA组细胞凋亡率下降(P0.05),细胞存活率明显增加(P0.01),G0/G1期细胞减少(P0.05)、S期细胞增加(P0.05),XPD、Bax表达降低(P均0.05),Bcl-2表达增高(P0.05)。结论 XPD能介导ox-LDL促进HUVEC凋亡作用。

Hxeroderma pigmentosum mediates the effect of ox-LDL on promoting the apoptosis of HUVEC

Aim To explore whether the hxeroderma pigmentosum D(XPD) could mediate the apoptosis of human umbilical vein endothelial cells (HUVEC) promoted by oxidized low density lipoprotein (ox-LDL). Methods The model of apoptosis of HUVEC was established by ox-LDL. XPD-siRNA was transfected into HUVEC with liposome,followed by treatment with ox-LDL. This experiment was divided into six groups:blank control group; negative control siRNA group; XPD-siRNA group; ox-LDL group; ox-LDL+negative control siRNA group; ox-LDL+XPD-siRNA group. Cell vitality was detected by MTT; Cell apoptosis rate and cell cycle were assessed with flow cytometry; The expressions of the XPD, Bax and Bcl-2 were detected by RT-PCR and Western blot. Results The optimal concentration of ox-LDL to establish the model of apoptosis of HUVEC was 100 mg/L. Compared with negative control siRNA group, the cell apoptosis rate of XPD-siRNA group was significantly decreased (P<0.05), the survival rate was significantly increased (P<0.01), the cell number in G0/G1 phase decreased (P<0.05), while increased in S phase (P<0.05), the expressions of XPD and Bax were declined (all P<0.05), while the expression of Bcl-2 was elevated (P<0.05); Compared with blank control group, the cell apoptosis rate of ox-LDL group increased significantly (P<0.01), the survival rate was decreased (P<0.05), the cell number in G0/G1 phase increased (P<0.05),while decreased significantly in S phase (P<0.01), the expressions of XPD and Bax were elevated (all P<0.05),while the expression of Bcl-2 was declined (P<0.05). Compared with ox-LDL+negative control siRNA group,the cell apoptosis rate of ox-LDL+XPD-siRNA group was reduced (P<0.05),the survival rate was increased(P<0.01), the cell number in G0/G1 phase decreased (P<0.05),while increased in S phase (P<0.05), the expressions of XPD and Bax were declined (all P<0.05), while the expression of Bcl-2 was elevated (P<0.05). Conclusion XPD can mediate the effect of ox-LDL on promoting the apoptosis of HUVEC.