G9a组蛋白甲基转移酶抑制剂BIX01294抑制血管平滑肌细胞增殖的机制研究  

Mechanism of G9a histone methyltransferase inhibitor BIX01294 inhibiting the proliferation of vascular smooth muscle cell

作　　者：王泽澜 罗稳健 赵俊勇 罗小林 秦浙学 WANG Ze-lan;LUO Wen-jian;ZHAO Jun-yong;LUO Xiao-lin;QIN Zhe-xue(Department of Cardiovasology,the Second Affiliated Hospital of Army Medical University,Chongqing 400037,China)

机构地区：[1]陆军军医大学第二附属医院心血管内科,重庆400037

出　　处：《局解手术学杂志》2025年第4期279-283,共5页Journal of Regional Anatomy and Operative Surgery

基　　金：重庆市自然科学基金面上项目(cstc2021jcyj-msxmX0716)。

摘　　要：目的探讨G9a组蛋白甲基转移酶抑制剂BIX01294对血管平滑肌细胞(VSMC)增殖的影响及其潜在的作用机制。方法将12只SD大鼠分为对照组和治疗组,每组6只。大鼠使用戊巴比妥钠腹腔注射麻醉,消毒备皮后,暴露颈总动脉、颈内外动脉,结扎颈内外静脉远心端,夹闭颈总动脉近心端,剪开颈外动脉,插入球囊导管并加压阻断血流,抽出球囊后结扎颈外动脉近心端。治疗组抽出球囊后维持颈总动脉夹闭状态,使用100μmol/L BIX01294溶液灌注处理30 s后恢复血流;对照组使用PBS灌注30 s后恢复血流。将VSMC分为正常组与BIX01294组,正常组细胞在含2%胎牛血清的低糖培养基中培养处理;BIX01294组细胞在正常组基础上分别添加2.5、5.0、7.5、10.0μmol/L BIX01294共处理。分别通过CCK-8实验、EDU实验检测细胞活性、增殖能力,筛选合适浓度的BIX01294。TUNEL实验检测正常组和BIX01294组、对照组和治疗组细胞凋亡情况。Western blot检测正常组和BIX01294组细胞自噬与凋亡相关蛋白表达水平。结果与0μmol/L相比,经不同浓度BIX01294处理后,VSMC的活性与增殖能力均呈浓度依赖性降低(P<0.05)。与正常组比较,BIX01294组VSMC凋亡水平增加(P<0.05),VSMC自噬及凋亡相关蛋白LC3Ⅰ/Ⅱ、Bax表达上调(P<0.05),Bcl-2、p62蛋白表达下调(P<0.05)。在体试验结果表明,BIX01294局部灌注加重了颈动脉VSMC的凋亡。结论BIX01294激活VSMC自噬与凋亡并抑制了VSMC的增殖。Objective To investigate the effect of G9a histone methyltransferase inhibitor BIX01294 on the proliferation of vascular smooth muscle cell(VSMC)and its underlying mechanism.Methods Twelve SD rats were divided into the control group and the treatment group,with 6 rats in each group.The rats were anesthetized by intraperitoneal injection with sodium pentobarbital,then exposed the common carotid artery and the internal and external carotid artery after disinfection and skin preparation,ligated the distal end of the internal and external jugular veins,clamped the proximal end of the common carotid artery,cut the external carotid artery,inserted the balloon catheter,blocked the blood flow by compression,and ligated the proximal end of the external carotid artery after withdrawing the balloon.The common carotid artery clamping state of rats in the treatment group was maintained after withdrawing the balloon,then restored the blood flow after perfusion with 100μmol/L BIX01294 solution for 30 seconds;rats in the control group were restored the blood flow after perfusion with PBS for 30 seconds.VSMC were divided into the normal group and the BIX01294 groups.Cells in the normal group were cultured in low glucose medium containing 2%fetal bovine serum,and cells in the BIX01294 groups were co-treated with 2.5,5.0,7.5 and 10.0μmol/L BIX01294 on the basis of the normal group,respectively.CCK-8 assay and EDU assay were used to detect the cell activity and proliferative ability respectively,to screen appropriate concentration of BIX01294.The cell apoptosis was detected by TUNEL assay between the normal group and the BIX01294 group,and the control group and the treatment group.Western blot was used to detect the expression levels of autophagy-and apoptosis-related proteins in the normal group and BIX01294 group.Results Compared with 0μmol/L,the activity and proliferative ability of VSMC decreased in a concentration-dependent manner after treatment with BIX01294 at different concentrations(P<0.05).Compared with the normal grou

关 键 词：G9a 血管平滑肌细胞 自噬 细胞凋亡 血管再狭窄 

分 类 号：R714.252[医药卫生—妇产科学]