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山东大学学报(医学版) ›› 2010, Vol. 48 ›› Issue (7): 37-41.

• 论文 • 上一篇    下一篇

内皮细胞类表皮生长因子域7对血管平滑肌细胞凋亡的影响

蒋卫东1,曾季平2,王欣1,鹿庆华1,秦爱琼1,刘玉胜1,吴伟芳2,葛志明3   

  1. 山东大学 1.第二医院心内科, 济南 250033;2.医学院生物化学与分子生物学研究所, 济南 250012;
    3.齐鲁医院心内科, 济南 250012
  • 收稿日期:2010-01-14 出版日期:2010-07-16 发布日期:2010-07-16
  • 基金资助:

    山东省自然科学基金资助项目(Y2006C59);山东省优秀中青年科学家科研奖励基金资助项目(2007BS03046)

Epidermal growth factor-like domain 7 regulating apoptosis of human aortic smooth muscle cells

JIANG Weidong1, ZENG Jiping2, WANG Xin1, LU Qinghua1, QIN Aiqiong1, LIU Yusheng1, WU Weifang2, GE Zhiming3   

  1. 1. Department of Cardiology, The Second Hospital of Shandong University, Jinan 250033, China;
    2. Institute of Biochemistry and Molecular Biology, School of Medicine, Shandong University, Jinan 250012, China;
    3. Department of Cardiology, Qilu Hospital of Shandong University, Jinan 250012, China
  • Received:2010-01-14 Online:2010-07-16 Published:2010-07-16

摘要:

目的   探讨人脐静脉内皮细胞株(HUVEC)中类表皮生长因子功能域7(EGFL7)对共培养的人主动脉平滑肌细胞(AoSMC)凋亡的影响以及可能的信号转导通路。方法   利用细胞共培养池行HUVEC与AoSMC共培养,将EGFL7的特异siRNA转染HUVEC细胞,利用MTS法检测HUVEC中EGFL7基因的有效沉默对共培养的AoSMC细胞存活率的影响;并合成无关siRNA作为阴性对照组(neg组),以未转染siRNA的细胞为空白对照组(con组)。同时利用分光光度法检测其对AoSMC细胞乳酸脱氢酶(LDH)、三磷酸腺苷(ATP)以及细胞凋亡蛋白酶(Caspase-3 )表达的影响;应用RT-PCR检测共培养AoSMC 的bcl-2及bax mRNA表达水平的改变。结果  与neg组及con组相比,干扰HUVEC细胞EGFL7表达12h后,AoSMC存活率无明显变化(P>0.05),而干扰24、36、48h后,存活率显著降低(P<0.05);干扰12、24、36、48h后,AoSMC细胞线粒体ATP释放量减少、培养基中LDH含量及Caspase-3表达量均增加(P<0.05)。RT-PCR检测结果显示,与neg组及con组相比,干扰siRNA转染HUVEC细胞24、36、48h,共培养的AoSMC凋亡基因bcl2表达水平轻度降低(P<0.05);但Bax表达水平在相应干预时间点均无明显改变(P>0.05)。结论  内皮细胞株EGFL7基因沉默时,可导致平滑肌细胞凋亡;EGFL7通过bcl-2相关信号通路调节SMC凋亡,而与Bax基因表达无关。

关键词: 类表皮生长因子功能域7;RNA干扰;平滑肌细胞;细胞凋亡;动脉硬化

Abstract:

Objective   To investigate the effect of epidermal growth factor (EGF)-like domain 7 (EGFL7) of human umbilical vein endothelial cells (HUVEC) on apoptosis of cocultured human aortic smooth muscle cells (AoSMC) and its signal transduction pathway. Methods   HUVEC and AoSMC were co-cultured in cell co-culture pools. The special siRNA targeting EGFL7 was transfected into HUVEC by lipofectamine, irrelevant siRNA as the negative group(neg) and siRNA-nontransfected cells as the control group(con).When the Egfl7 gene was silenced in HUVEC, AoSMC cell survival rates were determined by MTS, LDH, ATP and Caspase-3 release amount by spectrophotometry, and Bcl-2 and bax expressions in AoSMC by RT-PCR. Results   AoSMC survival rates decreased significantly after 24,36 and 48 hours(P<0.05). There were significant differences in ATP release amount, LDH content in medium and Caspase-3amount at 12, 24, 36 and 48 hours(P<0.05). The bcl-2 expression was gently reduced and there were significant differences at 24, 36 and 48 hours compared with the control and negative group(P<0.05). While there was no significant difference in the bax expression(P>0.05). Conclusion   The silence of EGFL7 in HUVEC could promote apoptosis of AoSMC. EGFL7 may regulate apoptosis of SMC via Bcl-2-related signal pathway.

Key words: Epidermal growth factorlike domain 7; RNA interference; Smooth Muscle Cell; Apoptosis; Atherosclerosis

中图分类号: 

  • 541.4
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