网膜素改善氧化应激对人动脉血管平滑肌细胞Ⅰ、Ⅳ型胶原表达的抑制作用
作者:
作者单位:

(1.泰达国际心血管病医院内分泌科,;2.天津国际生物医药联合研究院,天津市300457)

作者简介:

郑辉,硕士,副主任医师,从事糖尿病与动脉粥样硬化发生机制的研究,E-mail为zhui0123@163.com。刘慧娟,硕士,主要从事生物化学研究。

基金项目:

天津市滨海新区卫生局科技项目(2013BWK2006)


Omentin Attenues Oxidative Stress-induced Down-regulation of Ⅰ/Ⅳ Collagen in the Human Aortic Smooth Muscle Cells
Author:
Affiliation:

1.Department of Endocrine, TEDA International Cardiovascular Disease Hospital,Tianjin 300457,China;2.Tianjin International Joint Academy of Biotechnology & Medicine,Tianjin 300457,China)

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    摘要:

    目的 研究网膜素对氧化应激作用下的人主动脉平滑肌细胞胶原Ⅰ、Ⅳ表达的影响。方法 将人主动脉平滑肌细胞系培养,细胞密度到达90%以上后,随机分为5组:对照组、氧化应激组、网膜素组、细胞外调节蛋白激酶(ERK)抑制剂组和丝裂原活化蛋白激酶(p38MAPK)抑制剂组。其中,对照组不增加任何处理,氧化应激组加入叔丁基氢过氧化物(t-BHP,87.1 μmol/L),网膜素组(87.1 μmol/L t-BHP+600 μg/L 网膜素)、ERK抑制剂组、p38MAPK抑制剂组为在网膜素组基础上分别加入ERK抑制剂(PD98059,60 μmol/L)和p38MAPK抑制剂(SB203580,100 μmol/L),应用Western blot法检测细胞中胶原Ⅰ、Ⅳ表达量。结果 应用MTT法,筛选t-BHP最佳作用浓度为87.1μmol/L;与对照组相比,氧化应激组人动脉平滑肌细胞内Ⅰ、Ⅳ型胶原表达显著下降(P<0.01);与氧化应激组比较,网膜素组Ⅰ、Ⅳ型胶原表达显著升高(P<0.01);与网膜素组比较,ERK抑制剂组和p38MAPK抑制剂组Ⅰ、Ⅳ型胶原表达均显著下降(P<0.05)。结论 网膜素能改善氧化应激对人动脉平滑肌细胞Ⅰ、Ⅳ型胶原表达抑制的作用,可能通过此机制促进动脉粥样硬化斑块的稳定,ERK/p38MAPK途径可能参与网膜素的信号传导。

    Abstract:

    Aim To study the influence of omentin on the expression of collagen Ⅰ/Ⅳin the human aortic smooth muscle cells subjected to oxidative stress. Methods Human aortic smooth muscle cells were cultured in vitro.When cell density reached above 90%, cells were randomly divided into 5 groups:control group, oxidative stress group, omentin group, ERK(extracellular regulated protein kinases) inhibitor group and p38MAPK(p38 mitogen-activated protein kinases) inhibitor group. Among them, the control group did not receive any treatment, oxidative stress group were treated with tert-butyl hydroperoxide(t-BHP, 87.1 μmol/L) to simulate the state of oxidative stress; omentin group were treated,except for 87.1 μmol/L t-BHP,with 600 μg/L omentin; except for t-BHP(87.1 μmol/L) and omentin(600 μg/L),ERK inhibitors(PD98059,60 μmol/L) and p38MAPK inhibitors(SB203580,100 μmol/L )were used in ERK inhibitor group and p38MAPK inhibitor group respectively. The expression of collagen Ⅰ/Ⅳ were detected by Western blot hybridization. Results MMT method was used to screen the optimal concentration and the best action time of t-BHP.(1)The optimal concentration was 87.1 μmol/L;(2)Compared with the control group, the expression of collagen Ⅰ/Ⅳin human aortic smooth muscle cells in oxidative stress group decreased significantly(P<0.01);(3)Compared with the oxidative stress group, the expression of type Ⅰ/Ⅳcollagen in human aortic smooth muscle cells increased significantly in omentin group(P<0.01);(4)Compared with the omentin group, the expression of collagen Ⅰ/Ⅳin ERK inhibitor group and p38MAPK inhibitor group were decreased significantly(P<0.05). Conclusion Omentin could improve oxidative stress-induced down-regulation of Ⅰ/Ⅳcollagen in the human aortic smooth muscle cells,and probably promote the stabilization of atherosclerotic plaque through the mechanism. ERK/p38MAPK signaling pathway may be involved in omentin effect in vivo.

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引用本文

郑辉,刘慧娟,葛焕琦,张春风,胡睿,李雪粉.网膜素改善氧化应激对人动脉血管平滑肌细胞Ⅰ、Ⅳ型胶原表达的抑制作用[J].中国动脉硬化杂志,2016,24(2):141~144.

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  • 收稿日期:2015-02-26
  • 最后修改日期:2015-05-11
  • 在线发布日期: 2016-06-30