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中国应用生理学杂志 ›› 2018, Vol. 34 ›› Issue (5): 404-407.doi: 10.12047/j.cjap.5624.2018.092

• 研究论文 • 上一篇    下一篇

剪切应力环境下的内皮祖细胞对肝星状细胞增殖和生物功能的影响

陈信信1, 张晓芸2, 丁玉真1, 李鑫2, 官秀梅2, 李宏2, 成敏2, 崔晓栋2   

  1. 1. 潍坊医学院 生物科学与技术学院, 山东 潍坊 261053;
    2. 潍坊医学院 临床医学院, 山东 潍坊 261053
  • 收稿日期:2017-08-28 修回日期:2018-02-02 出版日期:2018-09-28 发布日期:2019-02-21
  • 通讯作者: 成敏,Tel:0536-8462268;E-mail:xiaodongcui@wfmc.edu.cn;崔晓栋,Tel:0536-8462022;E-mail:mincheng@wfmc.edu.cn E-mail:xiaodongcui@wfmc.edu.cn;mincheng@wfmc.edu.cn
  • 基金资助:
    国家自然科学基金(31570941,81700406,31270993);山东省自然科学基金(ZR2014JL018,ZR2016CM20);山东省高等学校科技计划项目(J15LK08,J14LK59,J14LK12);国家大学生创新计划项目(201510438013);潍坊医学院大学生科技创新基金(KX2016021,KX2015013);山东省中医药科技发展计划项目(2015-239);山东省医药卫生科技发展计划项目资助(2016WS0667)

Effects of endothelial progenitor cells on proliferation and biological function of hepatic stellate cells under shear stress

CHEN Xin-xin1, ZHANG Xiao-yun2, DING Yu-zhen1, LI Xin2, GUAN Xiu-mei2, LI Hong2, CHENG Min2, CUI Xiao-dong2   

  1. 1. College of Biological Science and Technology, Weifang Medical University, Weifang 261053, China;
    2. Clinical Medical College, Weifang Medical University, Weifang 261053, China
  • Received:2017-08-28 Revised:2018-02-02 Online:2018-09-28 Published:2019-02-21
  • Supported by:
    国家自然科学基金(31570941,81700406,31270993);山东省自然科学基金(ZR2014JL018,ZR2016CM20);山东省高等学校科技计划项目(J15LK08,J14LK59,J14LK12);国家大学生创新计划项目(201510438013);潍坊医学院大学生科技创新基金(KX2016021,KX2015013);山东省中医药科技发展计划项目(2015-239);山东省医药卫生科技发展计划项目资助(2016WS0667)

摘要: 目的:研究流体剪切应力条件下的内皮祖细胞(EPCs)对肝星状细胞(HSCs)增殖、粘附、迁移、凋亡等生物学功能以及成纤维化因子α-平滑肌肌动蛋白(α-SMA)、胶原I (Col-I)、胶原III (Col-III)表达的影响。方法:将HSCs与EPCs分别接种于共培养小室的上层和下层,共培养24 h后,给EPCs细胞施加12 dyne/cm2剪切应力,持续24 h。消化细胞,采用CCK-8法检测HSCs的增殖;流式细胞术检测HSCs的凋亡率;细胞贴壁法检测HSCs的粘附功能;Boyden小室检测HSCs的迁移;荧光定量PCR法及Western blot分别检测HSCs的α-SMA、Col-I、Col-III mRNA和蛋白质的表达情况。结果:在剪切应力条件下,EPCs生态小境能明显抑制HSCs的增殖、粘附和迁移能力,促进HSCs凋亡,下调HSCs中Col-I、Col-III mRNA和蛋白质的表达。结论:在剪切应力条件下,EPCs生态小境对HSCs纤维化的发展具有一定抑制作用。

关键词: 内皮祖细胞, 肝星状细胞, 剪切应力, 共培养, 生态小境

Abstract: Objective: To investigate the effects of endothelial progenitor cells (EPCs) under shear stress on the biological function such as proliferation, adhesion, migration, apoptosis and expression of α-smooth muscle actin (α-SMA), collagen-I and collagen-Ⅲ of hepatic stellate cells (HSCs). Methods: HSCs and EPCs were inoculated into the upper and lower layers of the co-culture chamber respectively and co-incubated for 24 hours. Then, 12 dyne/cm2 shear stress was applied to EPCs cells for another 24 hours. After that, proliferation, adhesion, migration and apoptosis of HSCs were detected by cell counting kit-8 (CCK-8) kit, cell adherent assay, Boyden cell migration assay and flow cytometry respectively. Fluorescence quantitative PCR and Western blot were used to detect the mRNA and protein expression of alpha -SMA, collagen I and collagen-Ⅲ in HSCs. Results: Under shear stress, EPCs ecological niche could obviously inhibit the proliferation, adhesion and migration of HSCs, promote the apoptosis of HSCs, and down-regulate the mRNA and protein expression of collagen-I, collagen-Ⅲ in HSC cells. Conclusion: Under shear stress, EPCs ecological niche could inhibit the fibrosis development of HSCs to a certain extent.

Key words: endothelial progenitor cells, hepatic stellate cells, shear stress, co-culture, ecological niche

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