剪切修复基因XPD抑制Ox-LDL诱导人脐动脉平滑肌细胞的增殖

发布时间：2018-06-13 11:28

本文选题：着色性干皮病D组基因 + 氧化低密度脂蛋白　；参考：《中国病理生理杂志》2017年12期

【摘要】：目的:探讨剪切修复基因——着色性干皮病D组基因(XPD)在氧化低密度脂蛋白(Ox-LDL)促血管平滑肌细胞增殖中的作用及机制。方法:将重组质粒pEGFP-N2/XPD利用脂质体转染人脐动脉平滑肌细胞(HUASMCs),实验分为空白对照组、空载质粒pEGFP-N2组、重组质粒pEGFP-N2/XPD组、Ox-LDL组、Ox-LDL+pEGFP-N2组和Ox-LDL+pEGFP-N2/XPD组。用MTT法和Ed U法测定各组细胞的增殖率;流式细胞术检测各组细胞周期分布;利用Western blot法检测XPD、caspase-3、Bcl-2和Bax的蛋白水平。结果:Western blot实验结果发现,与空白对照组相比,pEGFP-N2/XPD组的XPD表达增加(P0.05),表明转染成功;MTT和Ed U检测结果显示,pEGFP-N2/XPD组的细胞增殖率较空白对照组降低(P0.05);与Ox-LDL组比较,Ox-LDL+pEGFP-N2/XPD组细胞增殖明显被抑制(P0.05)。流式细胞术的检测结果显示,与空白对照组比较,pEGFP-N2/XPD组的S期细胞比例明显减少(P0.05),G0/G1期细胞比例明显增多(P0.05);与Ox-LDL组比较,Ox-LDL+pEGFP-N2/XPD组的S期细胞比例减少(P0.05),G0/G1期细胞比例明显增多(P0.05)。Western blot结果显示,与对照组比较,pEGFP-N2/XPD组的cleaved caspase-3和Bax蛋白水平增加(P0.05),Bcl-2蛋白表达降低(P0.05);与Ox-LDL组比较,Ox-LDL+pEGFP-N2/XPD组的cleaved caspase-3和Bax蛋白水平增加(P0.01),Bcl-2蛋白表达降低(P0.05)。结论:XPD能抑制HUASMCs的增殖并促其凋亡,还能抑制Ox-LDL的促HUASMCs增殖作用,有可能成为抗动脉粥样硬化治疗的靶点。
[Abstract]:Aim: to investigate the role and mechanism of shearing repair gene (XPD) in proliferation of vascular smooth muscle cells (VSMCs) induced by oxidized low density lipoprotein (Ox-LDL). Methods: the recombinant plasmid pEGFP-N2 / XPD was transfected into human umbilical artery smooth muscle cells (HUASMCs) by liposome. The cells were divided into blank control group, pEGFP-N2 / XPD group, Ox-LDL pEGFP-N2 group and Ox-LDL pEGFP-N2 / XPD group. The cell proliferation rate was measured by MTT assay and Ed U method, the cell cycle distribution was detected by flow cytometry, and the protein levels of Bcl-2 and Bax of XPD-caspase-3 were detected by Western blot. Results: Western blot experiment found that, Compared with the control group, the expression of XPD increased in pEGFP-N2 / XPD group, which indicated that the proliferation rate of pEGFP-N _ 2 / XPD group was lower than that of the blank control group, and that of Ox-LDL pEGFP-N _ 2 / XPD group was significantly inhibited by P0.05. Flow cytometry showed that, Compared with the control group, the percentage of S phase cells in pEGFP-N _ 2 / XPD group was significantly lower than that in the control group. The proportion of cells in the G _ 0 / G _ 1 phase of P0.05 / G _ 1 phase was significantly increased. Compared with the Ox-LDL group, the proportion of S phase cells in the Ox-LDL pEGFP-N _ 2 / XPD group was significantly lower than that in the Ox-LDL group, and the proportion of cells in the G _ 0 / G _ 1 phase of the Ox-LDL pEGFP-N _ 2 / XPD group was significantly higher than that in the Ox-LDL group. Compared with the control group, the expression of cleaved caspase-3 and Bax protein in pEGFP-N _ 2 / XPD group was increased, and the expression of Bcl-2 protein was decreased in the pEGFP-N _ 2 / XPD group, and the expression of cleaved caspase-3 and Bax protein was decreased in the Ox-LDL pEGFP-N _ 2 / XPD group compared with that in the Ox-LDL group, and the expression of Bcl-2 protein was decreased in the pEGFP-N _ 2 / XPD group. Conclusion: XPD can inhibit the proliferation and apoptosis of HUASMCs, and inhibit the proliferation of HUASMCs induced by Ox-LDL, which may be the target of anti-atherosclerosis therapy.
【作者单位】：南昌大学第二附属医院心内科;
【基金】：江西省自然科学基金资助项目(No.2010GZY0254)
【分类号】：R543.5

【相似文献】