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剪切修复基因D通过下调mTOR/LOX-1抑制ox-LDL诱导的人脐静脉平滑肌细胞增殖
作者:
作者单位:

(南昌大学第二附属医院心内科,江西省南昌市 330006)

作者简介:

余卫英,硕士研究生,研究方向为动脉粥样硬化和内科护理,E-mail:425599293@qq.com。通信作者夏子荣,博士,副主任医师,研究方向为动脉粥样硬化和心律失常,E-mail:xiazirong@sina.com。

基金项目:

江西省自然科学基金资助项目(2019GZY0254)；江西省卫生健康委科技计划(20204238)


Xeroderma pigmentosum group D gene inhibits the proliferation of human umbilical vein smooth muscle cell induced by ox-LDLvia down-regulating mTOR/LOX-1 pathway
Author:
Affiliation:

Department of Cardiovascular Medicine, the Second Affiliated Hospital of Nanchang Universty, Nanchang, Jiangxi 330006, China)

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    摘要:

    目的]探讨剪切修复基因D(XPD)在氧化型低密度脂蛋白(ox-LDL)促血管平滑肌细胞增殖中的作用及分子机制。 [方法]将重组质粒pEGFP-N2/XPD利用脂质体转染人脐静脉平滑肌细胞(HUVSMC),沉默哺乳动物雷帕霉素靶蛋白(mTOR)基因,用MTT、EdU法测定各组细胞的增殖；流式细胞仪检测各组凋亡率；利用Western blot检测XPD、血凝素样氧化型低密度脂蛋白受体1(LOX-1)、mTOR、p-mTOR、Bcl-2及Bax的表达。 [结果]与对照组比较,ox-LDL组XPD表达明显下调(P＜0.05),Bax蛋白表达下调(P＜0.05),Bcl-2蛋白、Bcl-2/Bax、mTOR磷酸化活性及LOX-1蛋白表达升高(P＜0.05)；MTT、BdU结果显示,ox-LDL组细胞增殖较对照组上调(P＜0.05)。转染pEGFP-N2/XPD重组质粒能上调Bax蛋白表达(P＜0.05)并抑制Bcl-2蛋白、mTOR磷酸化活性及LOX-1蛋白表达(P＜0.05),Bcl-2/Bax比值下调(P＜0.05)；流式细胞仪检测结果显示,转染pEGFP-N2/XPD重组质粒后,细胞凋亡率较对照组增加(P＜0.05)；MTT、BdU结果显示,转染pEGFP-N2/XPD重组质粒后,细胞增殖较对照组下调(P＜0.05)。沉默mTOR基因后,与对照组相比,siRNA mTOR组HUVSMC中LOX-1蛋白表达被抑制(P＜0.05)。 [结论]XPD能抑制HUVSMC的增殖并且促进其凋亡,下调mTOR和LOX-1蛋白表达,抑制ox-LDL的促HUVSMC增殖和抗凋亡作用,有可能成为抗动脉粥样硬化治疗的靶点。

    Abstract:

    Aim To investigate the mechanism and signal pathways of xeroderma pigmentosum group D(XPD) gene on the proliferation of human umbilical vein smooth muscle cell (HUVSMC) induced by ox-LDL. Methods HUVSMCs were transfected with the plasmids of pEGFP-N2/XPD using Lipofectamine 2000, and subsequently silent mTOR gene. MTT and EdU assay was used to detect the cell proliferation. Flow cytometry was used to examine the cell apoptosis. The expression of XPD, lectin-like oxidized low-density lipoprotein receptor 1(LOX-1), mTOR, phospho-mTOR, Bcl-2 and Bax was measured by Western blot. Results The expression of XPD and Bax protein was down-regulated in ox-LDL group (P＜0.05), while the expression of LOX-1, mTOR, Bcl-2 protein and the ratio of Bcl-2/Bax was significantly up-regulated (P＜0.05), compared with control group. Cell proliferation of ox-LDL group increased obviously (P＜0.05). After transfected with the pEGFP-N2/XPD plasmid, the expression of Bax was significantly up-regulated, while the expression of LOX-1, mTOR, Bcl-2 and the ratio of Bcl-2/Bax were significantly down-regulated (P＜0.05). Flow cytometry showed that overexpression of XPD increased the apoptosis rate of HUVSMC (P＜0.05). MTT and BdU showed that cell proliferation of pEGFP-N2/XPD group reduced compared with control group (P＜0.05). Compared with control group, the expression of LOX-1 was significantly down-regulated in siRNA mTOR group (P＜0.05). Conclusion XPD can inhibit HUVSMC proliferation and promote its apoptosis, and reduce the effect of ox-LDL promoting proliferation of HUVSMC via the mTOR/LOX-1 pathway. XPD may be the target of treatment of atherosclerosis.

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余卫英,夏子荣,李青,夏珍,李菊香.剪切修复基因D通过下调mTOR/LOX-1抑制ox-LDL诱导的人脐静脉平滑肌细胞增殖[J].中国动脉硬化杂志,2023,31(11):929~937.

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  • 收稿日期:2023-04-03
  • 最后修改日期:2023-07-02
  • 在线发布日期: 2023-12-05

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