【摘要】 目的探讨氧化性低密度脂蛋白（oxidized low density lipoprotein,ox-LDL）促进肝癌HepG2细胞中胆固醇蓄积的最适浓度和最适作用时间。方法将肝癌Hep G2细胞接种于含10%的胎牛血清和1%双抗（青霉素、链霉素）的高糖DMEM培养基中,置于5%浓度的CO₂培养箱中培养,每3天进行一次细胞传代,取对数增长期的细胞用于实验研究。实验分为5组,空白对照组和ox-LDL组（75 mg/L、50 mg/L、25 mg/L、12.5 mg/L）。每组9个样,分3个小组,每个小组3个样,3个小组分别培养24 h、48 h和72 h后测定细胞内胆固醇的积累并用油红O染色法定性观察。应用统计学软件SPSS Statistics 17.0进行t检验分析,数据以均数±标准差表示,对胆固醇定量检测结果进行统计分析,当P<0.05时认为差异具有统计学意义。结果与空白对照组比较,经过ox-LDL不同浓度和不同时间诱导后,总胆固醇含量（totalcholesterol,TC）、游离胆固醇含量（free cholesterol,FC）及胆固醇酯与总胆固醇比值（the ratio of cholesterol ester and totalcholesterol,CE/TC）均有一定程度的升高。尤以50 mg/Lox-LDL诱导48 h时[空白对照组:TC（98.21±2.85）mmol/L、FC（56.25±0.55）mmol/L、CE/TC 42.64±1.50;ox-LDL组:TC（146.7±3.59）mmol/L、FC（79.9±0.55）mmol/L、CE/TC 49.9±3.49]变化最明显。与空白对照组比较,ox-LDL组胞浆内有不同程度红色脂滴存在,且发现ox-LDL 50 mg/L作用48 h时,因含有过多脂滴而形成了红色聚集的脂团,而未加ox-LDL的空白对照组细胞内无明显脂滴颗粒。结论 ox-LDL不同诱导浓度和不同诱导时间对肝癌HepG2细胞内胆固醇（TC、FC、CE/TC）积累都有促进作用,而ox-LDL对肝癌HepG2细胞内胆固醇积累的影响最适浓度和时间为50 mg/L诱导48 h。更多还原

【Abstract】 Objective To investigate the oxidized low density lipoprotein（ox-LDL） promote hepatocellular carcinomaHepG2 cells accumulation of cholesterol in the optimum concentration and optimal time. Methods HepG2 cells wereinoculated with 10% fetal bovine serum and 1% dual-resistant（penicillin, streptomycin）high sugar DMEM medium, and werecultured in CO₂ culture tank of 5% concentration. Every 3 days for cells to extend, the exponential growth of the cells used inexperimental study. The experiment were divided into 5 groups,blank control group and ox-LDL groups（75 mg/L, 50 mg/L, 25 mg/L, 12.5 mg/L）. Each group of 9 samples were divided into 3 subgroups, 3 samples of each subgroup, 3 subgroups werecultured for 24 h, 48 h and 72 h respectively, and the accumulation of intracellular cholesterol was determined, and the color ofred O staining was observed. Application of statistical software Statistics 17.0 for t test analysis. Data formed as mean ±standard deviation, quantitative statistical was analyzed of test on cholesterol when P<0.05 for the difference was statisticallysignificance. Results Compared with the blank control group,after the ox-LDL induction of different concentration anddifferent time, total cholesterol（TC）, free cholesterol（FC） and the ratio of cholesterol ester and total cholesterol（CE/TC） had acertain degree of rise,especially the most obvious changes[Blank control group: TC（98.21±2.85）mmol/L, FC（56.25±0.55）mmol/L, CE/TC 42.64±1.50; Ox-LDL group: TC（146.7±3.59）mmol/L, FC（79.9±0.55）mmol/L, CE/TC 49.9±3.49] in the 50 mg/L ox-LDL cholesterol when induced 48 h. Compared with blank control group, ox-LDL group cytoplasm with red lipiddroplets of different levels, and found that it had much more fat droplets after 50 mg/L ox-LDL induced 48 h, and no significantlipid droplets were observed in the cell of the blank control group. Conclusion The different induction concentration of ox-LDL and the different induction time have the effect on the accumulation of cholesterol（TC, FC, CE/TC） in hepatocellular HepG2 cells,but the optimum concentration and optimal time of oxidized low density lipoprotein（ox-LDL） promoting theaccumulation of cholesterol is 50 mg/L, 48 hours..更多还原