组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2015, Vol. 11 ›› Issue (2): 95-109.doi: 10.3969/j.issn.1673-0364.2015.02.012

• 论著 • 上一篇    下一篇

坏死细胞对周围正常血管平滑肌细胞增殖迁移的影响及作用机制研究

孟亮，高远，陈宾，马坚，刘永，王微，赵雅琳   

  1. 沈阳医学院奉天医院；中国医科大学附属第一医院；沈阳市第四人民医院；沈阳医学院
  • 发布日期:2020-07-23

Effect of Necrotic Cells on Proliferation and Migration of Surrounding Normal Vascular Smooth Muscle Cells and the Underlying Mechanisms

MENG Liang,GAO Yuan,CHEN Bin,MA Jian,LIU Yong,WANG Wei,ZHAO Yalin   

  1. Fengtian hospital, Shenyang medical college;The First Hospital of China Medical University;Shenyang The Fourth Hospital of People;Shenyang medical college;
  • Published:2020-07-23
  • Contact: 辽宁省科学技术计划项目（2011225020）

摘要： 目的观察坏死的血管平滑肌细胞对周围正常细胞增殖和迁移能力的影响,同时探讨其可能的作用机制。方法采用无血清无糖低氧条件制备血管平滑肌细胞坏死模型,收集坏死细胞培养上清液干预正常细胞。实验设置无血清低糖DMEM培养的对照组、坏死上清(NCS)干预组、坏死上清与IL-1拮抗剂联合干预组、坏死上清与IL-6拮抗剂联合干预组、IL-1α干预组以及IL-6干预组。CCK-8方法检测细胞增殖能力变化,Transwell方法检测细胞迁移能力变化,RT-PCR方法检测各组细胞中IL-6和CRP m RNA表达情况,Western blot方法检测各组细胞CRP、Akt、p-Akt蛋白的表达。结果与对照组相比,NCS组、IL-1α组和IL-6组细胞的增殖能力和迁移能力均显著性增加(P<0.05),同时,NCS和IL-1α组血管平滑肌细胞IL-6表达升高(P<0.05),NCS组、IL-1α组和IL-6组细胞CRP和p-Akt的表达也呈增加趋势(P<0.05);而NCS+IL-1 RA组和NCS+IL-6 RA组细胞增殖、迁移能力较NCS组有所下降(P<0.05),CRP和pAkt的表达也相应有所降低(P<0.05)。结论坏死的血管平滑肌细胞可能通过释放IL-1α促进周围正常细胞IL-6的表达,进而上调CRP和p-Akt的表达,从而促进细胞增殖迁移。

关键词: 血管平滑肌细胞, 坏死, 增殖, 迁移

Abstract: Objective To observe the effect of necrotic vascular smooth muscle cells (VSMCs) on proliferation and migration of surrounding normal cells and to explore the possible mechanism. Methods VSMCs was cultured in oxygen-and glucose deprived medium and the supernatant was collected to intervene normally growing cells. The control group, NCS group, NCS+IL-1 RA group, NCS+IL-6 RA group, IL-1αgroup and IL-6 group were set in this experiment. CCK-8 and Transwell assays were used to determine the proliferation and migration of VSMCs. The mRNA expression of IL-6 and CRP were detected by RT-PCR, and the protein expression of CRP, Akt and p-Akt were evaluated by Western blot. Results Compared with the control group, the proliferation and migration ability of NCS group, IL-1αgroup and IL-6 group were significantly higher than that in the control group (P<0.05). Meanwhile, the expression of IL-6 were obviously increased in NCS group and IL-1αgroup (P<0.05), and the expression of CRP and p-Akt were also elevated in the three groups (P<0.05). Otherwise, the proliferation and migration ability of NCS+IL-1 RA group and NCS+IL-6 RA group were decreased and the expression of CRP and p-Akt were also declined (P<0.05). Conclusion Necrotic vascular smooth muscle cells may promoting the proliferation and migration of surrounding normal cells by inducing the expression of IL-6 and upregulating the expression of CRP and p-Akt.

Key words: Vascular smooth muscle cells, Necrosis, Proliferation,Migration

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