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氧化性低密度脂蛋白诱导浓度和时间对肝癌HepG2细胞胆固醇积累的影响

The impact that the induced concentration and time of ox-LDL on the accumulation of cholesterol in HepG2 cells
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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CO_2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。 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 CO_2 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 P0.05 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 chole
作者 李玉平 齐建彤 郑曙光 林昶 朱璨 柴艺汇 徐昌君 王和生 杨长福 LI Yuping, QI Jiantong, ZHENG Shuguang, LIN Chang, ZHU Can, CAI Yihui, XU Changjun, WANG Hesheng, YANG Changfu ( Guiyang College of Traditional Chinese Medicine, Guiyang, Guizhou 550002, China)
机构地区 贵阳中医学院
出处 《中国热带医学》 CAS 2018年第2期111-115,共5页 China Tropical Medicine
基金 贵阳中医学院2015年院内项目(No.042150032)
关键词 HEPG2细胞 氧化性低密度脂蛋白 胆固醇积累 动脉粥样硬化 HepG2 cells oxidized LDL cholesterol accumulation atherosclerosis
作者简介 李玉平(1990-),男,本科,助理实验师,研究方向:中药治疗肝代谢病。;杨长福,E-mail:yangchangfu@126.com
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