吉林大学学报(医学版)

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阿托伐他汀对同型半胱氨酸作用下人脐静脉内皮细胞Bcl-2启动子区甲基化水平的影响及其抗动脉粥样硬化机制

李录1,侯建军2,邱荣荣1,贾绍斌2,丛广志2,孙 娜3   

  1. 1.南京医科大学附属无锡市第二人民医院心内科,江苏 无锡 214002;2.宁夏医科大学总医院心脏中心心内科,宁夏 银川 750004;3.陕西省渭南市中心医院心内科,陕西 渭南 714000
  • 收稿日期:2013-09-04 出版日期:2014-09-28 发布日期:2014-11-24
  • 通讯作者: 贾绍斌 (Tel:0951-6743232,E-mail:jsbxn@163.com) E-mail:jsbxn@163.com
  • 作者简介:李 录(1978-),男,河北省张家口市人,医学硕士,主要从事冠状动脉粥样硬化发病机制的研究。
  • 基金资助:

    宁夏回族自治区科技厅自然科学基金资助课题(NZ11266)

Influence of atorvastatin in Bcl-2 methylation in cultured human umbilical endothelial cells treated with homocysteineand its mechanism of anti-arteriosclerosis

LI Lu1,HOU Jian-jun2,QIU Rong-rong1,JIA Shao-bin2,CONG Guang-zhi2,SUN Na3   

  1. 1. Department of Cardiology,Wuxi Second People’s Hospital,Nanjing Medical University,Wuxi 214002,China;2.Center of Cardiology, General Hospital,Ningxia Medical University,Yingchuan 750004,China;3.Department of Cardiology,Weinan Central Hospital,Shaanxi Province,Weina
    n 714000,China
  • Received:2013-09-04 Online:2014-09-28 Published:2014-11-24

摘要:

目的:探讨阿托伐他汀对同型半胱氨酸(Hcy)作用下人脐静脉内皮细胞(HUVECs)Bcl-2基因启动子区甲基化水平及其表达的影响,阐明阿托伐他汀抗动脉粥样硬化(AS)的可能机制。方法:HUVECs分别采用含有0、2、4、8、16和32  mmol/L Hcy的RPMI 1640培养液作用48 h,采用MTT实验检测细胞增殖抑制率及半数抑制浓度(IC50),根据IC50结果将本实验分为对照组(0 mmol/L Hcy)、Hcy组(9.00 mmol/L Hcy)、阿托伐他汀组(9.00 mmol/L Hcy+1×10-3 mmol/L阿托伐他汀)。各组细胞培养48 h后,流式细胞术检测细胞凋亡率,荧光定量PCR法检测Bcl-2 基因的mRNA表达,Western blotting 法检测Bcl-2蛋白表达水平,甲基化特异性PCR(MSP)联合巢式降落式PCR法检测Bcl-2启动子区甲基化水平。结果:与对照组比较,Hcy组细胞凋亡率上升(P<0.01), Bcl-2基因mRNA及蛋白表达水平下降(P<0.01),Bcl-2基因启动子区甲基化水平降低(P<0.01);与Hcy组比较,阿托伐他汀组内皮细胞凋亡率下降(P<0.01), Bcl-2基因mRNA及蛋白表达水平增加(P<0.05),Bcl-2基因启动子区甲基化水平升高(P<0.05)。结论:阿托伐他汀可通过调节Hcy作用下的HUVECs Bcl-2基因甲基化水平抑制细胞凋亡。

关键词: 阿托伐他汀, 动脉粥样硬化, 同型半胱氨酸, Bcl-2蛋白, DNA甲基化, 细胞凋亡

Abstract:

Abstract:Objective To investigate the influence of atorvastatin in methylation and expression level of Bcl-2 in human umbilical endothelial cells(HUVECs) treated with homocysteine(Hcy) and to expound potential mechanism of atorvastatin resisting arteriosclerosis.Methods After HUVECs were treated with 0,2,4,8,16,and 32 mmol/L  Hcy for 48 h,MTT was used to measure the inhibitory rates of HUVECs and the half inhibitory concentration (IC50).According to the experimental results,the HUVECs cultured in vitro were divided into control group(0.00 mmol/L Hcy),Hcy group(9.00 mmol/L Hcy),and atorvastatin group(9.00 mmol/L Hcy+1×10-3 mmol/L atorvastatin).After treated for 48 h,flow cytometry was used to detect the apoptotic rate of cells,the mRNA expression of Bcl-2 was analyzed by fluorescence quantitative PCR,the protein expression of Bcl-2 was detected by Western blotting method,and the methylation level of Bcl-2 promoter region was determined by nest touch-down PCR combined with methylation specific PCR(MSP).Results Compared with control group,the apoptotic rate of HUVECs in Hcy group was increased(P<0.01),the mRNA and protein expression levels of Bcl-2 were significantly decreased(P<0.01),and the Bcl-2 promoter region methylation level was also decreased(P<0.01).Compared with Hcy group,the apoptotic rate of HUVECs in atorvastatin group was decreased(P<0.01),the mRNA and protein expression levels of Bcl-2 gene were increased(P<0.05),and the Bcl-2 promoter region methylation level was also increased(P<0.05).Conclusion Atorvastatin can prevent the apoptosis of HUVECs induced by Hcy through regulating Bcl-2 methylation.

Key words: atorvastatin, atherosclerosis, homocysteine, Bcl-2 protein, DNA methylation, apoptosis

中图分类号: 

  • R543.5