橄榄苦苷对HepG2细胞脂肪变性的作用  

Role of oleuropein in the steatosis of HepG2 cells

作　　者：丁利敏 朴莲淑[1] 张孟 刘城 于庆功 DING Limin;PIAO Lianshu;ZHANG Meng;LIU Cheng;YU Qinggong(Department of Gastroenterology,Zhongshan Hospital Affiliated to Dalian University,Dalian 116001,China)

机构地区：[1]大连大学附属中山医院消化科,辽宁大连116001

出　　处：《中华实用诊断与治疗杂志》2018年第9期841-844,共4页Journal of Chinese Practical Diagnosis and Therapy

基　　金：国家自然科学基金(81301979)

摘　　要：目的探讨橄榄苦苷对HepG2细胞脂肪变性的影响。方法人肝癌HepG2细胞株分别用浓度0、0.5、1、1.5、2mmol/L游离脂肪酸(free fatty acid,FFA)培养24h,MTS法测定细胞生存率,尼罗红染色观察细胞内脂滴颗粒堆积情况,ELISA法检测细胞三酰甘油(triacylglycerol,TG)水平,选择1mmol/L FFA进行后续实验。取人肝癌HepG2细胞株随机分为空白组(培养液),FFA组(培养液+1 mmol/L FFA),1μmol/L橄榄苦苷组(培养液+1 mmol/L FFA+1μmol/L橄榄苦苷)、10μmol/L橄榄苦苷组(培养液+1mmol/L FFA+10μmol/L橄榄苦苷)、100μmol/L橄榄苦苷组(培养液+1mmol/L FFA+100μmol/L橄榄苦苷)、1 000μmol/L橄榄苦苷组(培养液+1mmol/L FFA+1 000μmol/L橄榄苦苷),培养24h采用MTS法测定细胞生存率,尼罗红染色观察细胞内脂滴颗粒堆积情况,ELISA法检测细胞TG水平。结果 MTS结果显示,FFA浓度为1.5、2mmol/L时细胞生存率[(66.50±5.83)%、(57.63±11.63)%]低于0mmol/L时(100%)(P<0.05),0.5、1mmol/L时细胞生存率[(95.96±3.50)%、(95.49±7.75)%]与0mmol/L比较差异无统计学意义(P>0.05);FFA浓度为0.5、1mmol/L时细胞中TG水平[(0.91±0.02)、(0.96±0.04)mmol/L]均高于0 mmol/L[(0.37±0.05)mmol/L](P<0.05);1、10、100μmol/L橄榄苦苷组细胞生存率[(96.22±8.05)%、(94.94±4.51)%、(97.10±2.53)%]与空白组(100%)、FFA组[(95.42±9.42)%]比较差异无统计学意义(P>0.05);1 000μmol/L橄榄苦苷组细胞生存率[(41.35±4.84)%]明显低于空白组、FFA组(P<0.05);10、100μmol/L橄榄苦苷组细胞中TG水平[(0.64±0.07)、(0.57±0.02)mmol/L]低于FFA组[(0.96±0.02)mmol/L](P<0.05),1μmol/L橄榄苦苷组细胞中TG水平[(0.92±0.05)mmol/L]与FFA组比较差异无统计学意义(P>0.05);显微镜下10、100μmol/L橄榄苦苷组脂肪颗粒较FFA组明显减少。结论 10、100μmol/L橄榄苦苷可减少HepG2细胞内TG生成,防止肝细胞脂肪变性。Objective To investigate the effect of oleuropein on the steatosis of HepG2 cells. Methods Human hepatoma HepG2 cells were cultured for 24 h with free fatty acid （FFA） at concentration of 0, 0. 5, 1, 1. 5 and 2 mmol/L, respectively. The cell survival rate was detected by MTS assay, and intracellular lipid droplets were observed by Nile red staining. The level of triacylglyeerol （TG） was determined by ELISA, and 1 mmol/L FFA was selected for subsequent experiments. Human hepatoma HepG2 cell line was randomly divided into blank group （culture medium）, FFA group （culture medium ＋ 1 mmol/L FFA）, 1μmol/L oleuropein group （culture medium - 1 mmol/L FFA＋ 1 μmol/L oleuropein）, 10 μmol/L oleuropein group （culture medium ＋ 1 mmol/L FFA ＋ 10 μmol/L oleuropein）, 100 μmol/L oleuropein group （culture solution ＋ 1 mmol/L FFA ＋100 μmol/L oleuropein）, 1 000 μmol/L oleuropein group （culture medium ＋ 1 mmol/L FFA ＋ 1 000 μmol/L oleuropein）, cultured for 24 h. The cell survival rate was determined by MTS method. The accumulation of internal lipid droplets was observed. The level of triacylglycerol （TG） was detected by ELISA. Results MTS results showed that the cell survival rate was significantly lower when the FFA concentration was 1. 5 mmol/L （（66. 50 ~ 5. 83）%） and 2 mmol/L （（57. 63 _＋ 11. 63）%） than that when FFA concentration was 0 mmol/L （100%） （P〈0.05）, and showed no significant difference when the FFA concentration was 0.5 mmol/L （（95. 96± 3. 50）%） and 1 mmol/L （（95. 49±7. 75）%） in comparison with that when the FFA concentration was 0 mmol/L （P〉0.05）. The TG level was significantly higher when FFA concentration was 0.5 mmol/L （（0.91±0.02） mmol/L） and 1 mmol/L （（0.96±0.04） mmol/L） than that when FFA concentration was 0 mmol/L （（0.37±0.05） mmol/L） （P〈0.05）. There were no significant differences in the cell survival rates in 1, 10 and 100 μmol/L oleuropein groups （（96.22±8.05）

关 键 词：非酒精性脂肪性肝病 橄榄苦苷 HEPG2细胞 三酰甘油 

分 类 号：R575.5[医药卫生—消化系统]