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As2O3对大鼠血管平滑肌细胞凋亡、PCNA表达以及表型的影响
Effect of As2O3 on Apoptosis, PCNA expression and Phenotype of Rat Vascular Smooth Muscle Cells
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DOI:
作者:
*田文杰 沈雳 杨巍 龚飞荣 陆浩 孙爱军 邹云增 葛均波
TIAN Wen-jie, SHEN Li, YANG Wei, GONG Fei-rong, LU Hao,SUN Ai-jun, ZOU Yun-zeng,GE Jun-bo
作者单位:
610072 成都,四川省医学科学院•四川省人民医院心内科(田文杰);200032 上海,复旦大学附属中山医院心内科 上海市心血管病研究所(葛均波 沈雳 陆浩 孙爱军 邹云增);150001 哈尔滨,哈尔滨医科大学附属第一医院心内科 黑龙江省心血管病研究所(杨巍);4200237华东理工大学高分子材料研究所(龚飞荣)
Department of Cardiology, Sichuan Academy of Medical sciences & Sichuan provincial people’s hospital, Chengdu, 610072,China(TIAN Wen-jie); Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China(SHEN Li,LU Hao,SHUN Ai-jun,ZOU Yun-zeng,GE Jun-bo);Key Laboratory for Ultrafine Materials of Ministry of Education, School of Materials Science and Engineering, East China University of Science and Technology, Shanghai 200237, China(GONG Fei-rong);Department of Cardiology, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, China(YANG Wei)
关键词:
三氧化二砷;凋亡;增殖细胞核抗原;表型;血管平滑肌细胞
Arsenic trioxide (As2O3); Apoptosis; Proliferating Cell Nuclear Antigen(PCNA); Phenotype; Vascular smooth muscle cells (VSMCs)
摘要:
【摘要】目的 观察As2O3对大鼠血管平滑肌细胞凋亡、PCNA表达以及表型转化的影响,为As2O3洗脱支架的应用提供生物学基础。方法 贴壁法培养大鼠胸主动脉平滑肌细胞,不同剂量As2O3作用48h后,以流式细胞仪分析平滑肌细胞凋亡率,Hoechest染色观察5umol/L As2O3作用不同时间细胞凋亡情况,以Western blot方法检测As2O3作用后PCNA蛋白的表达。采用胎牛血清和PDGF诱导血管平滑肌细胞由收缩型向合成型转化,观察As2O3对该表型转化过程的影响。结果 1、3、5umol/LAs2O3干预组的平滑肌细胞凋亡率分别为(15.1±3.3)%、(26.5±4.5)%、(39.3±6.6)%,较未干预组(5.2±1.1)%明显增加(P<0.01);Hoechest染色显示随As2O3干预时间的延长,细胞凋亡逐渐增多。Western blot检测表明As2O3使血管平滑肌细胞PCNA蛋白表达减少。胎牛血清和PDGF诱导VSMCs由收缩型向合成型转化,使收缩型平滑肌细胞表面标志α-actin表达减少,As2O3对这一转化过程没有影响,不能增加α-actin的表达量(P>0.05)。结论 As2O3剂量和时间依赖性的诱导大鼠VSMCs凋亡,能明显降低PCNA表达,但对血管平滑肌细胞表型转换无明显影响。
ABSTRACT Objective To investigate the effect of As2O3 on apoptosis,Proliferating Cell Nuclear Antigen(PCNA) expression and phenotype of rat Vascular Smooth Muscle Cells (VSMCs). Methods The ratio of apoptotic cells was detected by flow cytometry and the expression of PCNA was determined by Western blot after VSMCs was treated by As2O3 for 48 hours, and the apoptotic cells were abserved by Hoechest dying at different time point with the treatment of As2O3 at the concentration of 5umol/L. The phenotypic transformation from contractile type to synthetic type was induced by fetal bovine serum and PDGF, and the effect of As2O3 on α-actin protein was analyzed by Western blot. Results The ratio of apoptotic cells was (15.1±3.3)%, (26.5±4.5)%, (39.3±6.6)% respectively at the As2O3 concentration of 1, 3, 5umol/L, which was higher than that of As2O3 concentration of 0umol/L(p<0.01). Hoechest dying showed that apoptotic cells significantly increased with prolonging of treatment time at the As2O3 concentration of 5umol/L. PCNA expression obviously was inhibited with the concentration increase of As2O3. The decreasing α-actin expression was induced by fetal bovine serum and PDGF, and As2O3 had no effect on this process(p>0.05). Conclusions As2O3 dose- and time-dependently induced apoptosis of VSMCs with downreguration of PCNA expression, while didn’t increase α-actin expression which was inhibited by fetal bovine serum and PDGF, therefore had no effect on the phenotypic transformation of VSMCs.
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