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

doi:10.12047/j.cjap.5624.2018.092

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

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. 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. 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/Abstract

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

陈信信, 张晓芸, 丁玉真, 李鑫, 官秀梅, 李宏, 成敏, 崔晓栋. 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[J]. 中国应用生理学杂志, 2018, 34(5): 404-407.

CHEN Xin-xin, ZHANG Xiao-yun, DING Yu-zhen, LI Xin, GUAN Xiu-mei, LI Hong, CHENG Min, CUI Xiao-dong. Effects of endothelial progenitor cells on proliferation and biological function of hepatic stellate cells under shear stress[J]. CJAP, 2018, 34(5): 404-407.

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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

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

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