摘要
目的通过油酸(oleic acid,OA)诱导人肝癌细胞系BEL-7402建立非酒精性脂肪性肝炎(nonalcoholic steatohepatitis,NASH)的细胞模型,探讨肝刺激因子(hepatic stimulator substance,HSS)对NASH的保护作用。方法采用25、50、100、200、400μmol/L浓度OA处理BEL-7402细胞24h后,再采用噻唑蓝[3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide,MTT]法检测OA对细胞存活率的影响,使用脂质检测试剂盒测定细胞中三酰甘油(triglyceride,TG)的含量,以最适实验条件作用于转染空载体和稳定转染HSS的BEL-7402细胞,检测细胞内TG的含量及细胞上清中丙氨酸转氨酶(alanine aminotransferase,ALT)含量,油红O染色观察细胞内脂质堆积情况以及使用caspase-3/7活性测定试剂盒检测细胞凋亡情况,综合观察稳定转染HSS保护肝细胞免受损伤的能力。结果用200和400μmol/L浓度的OA作用于BEL-7402细胞24h可以使细胞活力显著降低。OA浓度为100μmol/L时,BEL-7402细胞内的TG含量最多。与转染空载体的细胞相比,稳定转染HSS细胞内的TG含量和胞外ALT含量均显著下降,用油红O染色可以观察到细胞内红色脂滴沉积减少。结论 HSS对OA诱导的NASH肝损伤起保护作用。
Objective To investigate the protective effects of hepatic stimulator substance（HSS）on non-alcoholic steatohepatitis（NASH）by establishing a BEL-7402 cell model of NASH induced by oleic acid（OA）.Methods After OA treatment at different concentrations（25,50,100,200,400μmol/L）for 24 h,BEL-7402 cells viability effects were measured using 3-（4,5）-dimethylthiahiazo（-z-y1）-3,5-di- phenytetrazoliumromide（MTT）.And the cellular triglyceride（TG）contents were determined by assay kits.The HSS-expressing BEL-7402 cells and vectortransfected BEL-7402 cells were treated with 100μmol/L OA for 24 h.The cellular TG contents,alanine aminotransferase（ALT）and caspase-3/7activity were determined respectively.The lipid drops were assessed by oil red O staining.Results The cell viability was significantly reduced by OA treatment 200 and 400μmol/L for 24 hin BEL-7402 cells.When the OA concentration was100μmol/L,the TG contents of BEL-7402 cells were maximum.The TG contents,lipid accumulation and the cellular supernatant levels of ALT in HSS-expressing BEL-7402 cells were less than those in the control vector-transfected BEL-7402 cells.Conclusion HSS can protect hepatocyte from NASH induced by OA treatment to BEL-7402 cells.
出处
《河北医科大学学报》
CAS
2015年第5期501-504,共4页
Journal of Hebei Medical University
基金
国家自然科学基金资助项目(31371169)
关键词
脂肪肝
肝刺激因子
油酸
fatty liver
hepatic stimulator substance
oleic acid
