CAJ | 학술논문

目的利用体外脂肪变性的模型,探讨不同自由脂肪酸(FFA)的细胞毒性和诱导细胞凋亡的机制.方法分别采用DAPI核染剂和借助半胱氨酸天冬氨酸蛋白酶3(Caspase3)荧光分析试剂盒检测人肝肿瘤细胞株HepG2在不同脂肪酸在不同时间后(12 h、24 h、48 h)TG和细胞凋亡率和Cagpage3活性的变化,以及Z-VAD-FMK在不同时间对棕榈酸(PA)诱导的细胞凋亡和Caspagc3的活性的抑制作用.结果亚油酸(LA)组、油酸(OA)组在各时间段的TG的积累高于PA组,但凋亡率和Cagpage3活性均低于PA组,并呈时间依赖性.加入Z-VAD-FMK后PA导致的凋亡率和Cmpage3活性均显著性降低.结论棕榈酸以Caspage3蛋白酶依赖的方式诱导脂凋亡.油酸和亚油酸通过形成甘油三酯,防止肝细胞脂凋亡.
목적 이용체외지방변성적모형,탐토불동자유지방산(FFA)적세포독성화유도세포조망적궤제.방법 분별채용DAPI핵염제화차조반광안산천동안산단백매3(Caspase3)형광분석시제합검측인간종류세포주HepG2재불동지방산재불동시간후(12 h、24 h、48 h)TG화세포조망솔화Cagpage3활성적변화,이급Z-VAD-FMK재불동시간대종려산(PA)유도적세포조망화Caspagc3적활성적억제작용.결과 아유산(LA)조、유산(OA)조재각시간단적TG적적루고우PA조,단조망솔화Cagpage3활성균저우PA조,병정시간의뢰성.가입Z-VAD-FMK후PA도치적조망솔화Cmpage3활성균현저성강저.결론 종려산이Caspage3단백매의뢰적방식유도지조망.유산화아유산통과형성감유삼지,방지간세포지조망.
Objective To investigate the cytotoxicity of different kind of free fatty acids FFA and the possible mechanism induced apoptosis by liver tumor HepG2 cell line through in vitro Steatosis model. Methods A HepC2 model of NASH was incubated with palmtic acid PA, linoleic acid LA or oleic acid OA for a period of 12h, 24h or 48h. Concentration of TG in the supernatant was measured by an enzymatic assay kit from Stanbio. Nuclear staining with 4, 6-diamidino-2-phenylindole DAPI and fluorescence microscopy were used to quantitate apoptotic cells. Caspase 3 activity was evaluated by active caspase 3 apoptosis kit with flow cytometry. At the same time, apop-totic cells and caspase 3 activity were measured by adding the pancaspase inhibitor Z-VAD-fmk. Results TG of OA and LA was more than PA, while apoptotic and caspase3 activity of OA and LA was lower than PA in a time-dependent manner. Z-VAD-fmk attenuated apoptosis and caspase 3 activity by PA in a time-dependent manner. Conclusion The results demonstrated that palmtic acid mediated lipoapoptosis occurs by a caspase-dependent mechanism. It showed that transformation OA and LA to TG reduced lipoapoptosis.