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/cm^2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。
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/cm^2 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.
作者
陈信信
张晓芸
丁玉真
李鑫
官秀梅
李宏
成敏
崔晓栋
CHEN Xin-xin;ZHANG Xiao-yun;DING Yu-zhen;LI Xin;GUAN Xiu-mei;LI Hong;CHENG Min;CUI Xiao-dong(College of Biological Science and Technology,Weifang Medical University,Weifang 261053,China;Clinical Medical College,Weifang Medical University,Weifang 261053,China)
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2018年第5期404-407,共4页
Chinese Journal of Applied Physiology
基金
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)
关键词
内皮祖细胞
肝星状细胞
剪切应力
共培养
生态小境
endothelial progenitor cells
hepatic stellate cells
shear stress
co-culture
ecological niche
作者简介
通讯作者:成敏,E-mail:mincheng@wfmc.edu.cn;通讯作者:崔晓栋,E-mail:xiaodongcui@wfmc.edu.cn
