CAJ | 학술논문

目的:肝星形细胞(hepatic stellate cells,HSCs)是参与肝纤维化和肝硬化发展过程的主要细胞.在肝纤维化过程中,肝星形细胞增殖并发生表型转化,从静止状态向肌纤维母细胞样转化.后者的归宿可为凋亡,也可重归静止状态.目前转化肌纤维母细胞样细胞的凋亡机制尚未明确.本文研究细胞外信号调节激酶(extracellular signalregulated kinases,ERKs)磷酸化状态对十字孢碱诱导HSCs凋亡的影响.方法:采用Western印迹和流式细胞术检测4种肝星形细胞株(CFSC-8B,-2G,-3H和-5H)的ERKs表达水平和细胞凋亡状态.结果:4种肝星形细胞株各具有形态特异性,并与其内α-SMA表达水平相符,其中CFSC-8B细胞株α-SMA表达水平为最高.虽然ERK1/2总蛋白表达水平在4种细胞株相似,但磷酸化ERK1/2在CFSC-8B和CFSC-2G两个细胞株中表达明显高于CFSC-3H和CFSC-5H细胞株.进一步采用CFSC-8B细胞(ERK1/2高磷酸化水平)和CFSC-5H细胞(ERK 1/2低磷酸化水平),通过十字孢碱诱导细胞凋亡.结果显示CFSC-8B细胞对十字孢碱诱导的细胞凋亡敏感性明显增加.同时,十字孢碱还可进一步增加这2株细胞内ERK1/2的磷酸化程度.结论:HSCs中ERK1/2的磷酸化程度决定细胞对十字孢碱所致细胞凋亡的敏感性.
목적:간성형세포(hepatic stellate cells,HSCs)시삼여간섬유화화간경화발전과정적주요세포.재간섬유화과정중,간성형세포증식병발생표형전화,종정지상태향기섬유모세포양전화.후자적귀숙가위조망,야가중귀정지상태.목전전화기섬유모세포양세포적조망궤제상미명학.본문연구세포외신호조절격매(extracellular signalregulated kinases,ERKs)린산화상태대십자포감유도HSCs조망적영향.방법:채용Western인적화류식세포술검측4충간성형세포주(CFSC-8B,-2G,-3H화-5H)적ERKs표체수평화세포조망상태.결과:4충간성형세포주각구유형태특이성,병여기내α-SMA표체수평상부,기중CFSC-8B세포주α-SMA표체수평위최고.수연ERK1/2총단백표체수평재4충세포주상사,단린산화ERK1/2재CFSC-8B화CFSC-2G량개세포주중표체명현고우CFSC-3H화CFSC-5H세포주.진일보채용CFSC-8B세포(ERK1/2고린산화수평)화CFSC-5H세포(ERK 1/2저린산화수평),통과십자포감유도세포조망.결과현시CFSC-8B세포대십자포감유도적세포조망민감성명현증가.동시,십자포감환가진일보증가저2주세포내ERK1/2적린산화정도.결론:HSCs중ERK1/2적린산화정도결정세포대십자포감소치세포조망적민감성.
Objective:Hepatic stellate cells (HSCs) are the principal cells responsible for the development of hepatic fibrosis and cirrhosis.During the fibrotic process,HSCs undergo proliferation and transdifferentiation from a quiescent to myofibroblast-like phenotype.The fate of myofibroblastlike HSCs includes apoptosis or reversion back to a quiescent phenotype.The mechanisms involved in the apoptotic process of HSCs have yet to be determined.The purpose of the present study is to determine the effects of extracellular signal-regulated kinases (ERKs) phosphorylation on the apoptosis of HSCs induced by staurosporine.Methods:We used Western blot and flow cytometry to detect the expression level of ERK and cell apoptosis status in four rat hepatic stellate cell lines (CFSC-8B,-2G,-3H and -5H).Results:Each hepatic stellate cell line had a distinct morphology consistent with their expression level of α-SMA and that CFSC-8B cells had the highest α-SMA expression.Although all four cell types expressed similar levels of ERK1/2,phosphorylation levels were significantly higher in CFSC8B and CFSC-2G than in CFSC-3H and CFSC-5H cells.When CFSC-8B ceils (high ERK1/2 phosphorylation) and CFSC-5H cells (low ERK1/2 phosphorylation) were employed to examine staurosporine-induced apoptosis,CFSC-8B cells were significantly more sensitive.Staurosporine further increased ERK1/2 phosphorylation in both cell lines.Conclusion:ERK1/2 phosphorylation in HSCs determines the sensitivity of HSCs to staurosporine-induced apoptosis.