油酸钠致L02肝细胞脂肪变性和炎症效应  

Steatosis and inflammation of L02 hepatocytes induced by sodium oleate

作　　者：陈晨 马妍 公维一 张虎 王晶波 杨倬 秦文 李岩 卓勤 贾旭东 霍军生 Chen Chen;Ma Yan;Gong Weiyi;Zhang Hu;Wang Jingbo;Yang Zhuo;Qin Wen;Li Yan;Zhuo Qin;Jia Xudong;Huo Junsheng(Key Laboratory of Trace Element Nutrition of National Health Commission(NHC),National Institute for Nutrition and Health,Chinese Center for Disease Control and Prevention,Beijing 100050,China;The People’s Hospital of Xintai,Tai'an 271200,China;Key Laboratory of Food Safety Risk Assessment of National Health Commission,China National Center for Food Safety Risk Assessment,Beijing 100021,China)

机构地区：[1]中国疾病预防控制中心营养与健康所国家卫生健康委员会微量元素营养重点实验室,北京100050 [2]山东省新泰市人民医院,泰安271200 [3]国家食品安全风险评估中心,国家卫生健康委员会食品安全风险评估重点实验室,北京100021

出　　处：《卫生研究》2020年第3期381-385,共5页Journal of Hygiene Research

基　　金：国家重点研发计划(No.2018YFC1602104)。

摘　　要：目的观察油酸钠导致的L02肝细胞脂肪变性和炎症效应。方法不同浓度油酸钠孵育L02肝细胞24 h,检测细胞活性。选定75、150和300μmol/L油酸钠分别与L02肝细胞培养24 h,油红染色观察细胞脂质蓄积情况,检测细胞中甘油三酯含量和上清液中白介素6(interleukin 6,IL-6)含量,免疫印迹法检测沉默信息调节因子1(silent information regulator 1,SIRT1)和核因子-κB(nuclear factorκB,NF-κB)的表达,流式细胞仪检测细胞表面Toll样受体2(Toll-like receptor,TLR2)和TLR4的表达。结果随着油酸钠浓度增加,细胞活性下降,细胞生长抑制率升高。75、150和300μmol/L油酸钠干预L02肝细胞甘油三酯含量明显高于对照组(P<0.01、P<0.001和P<0.001),上清液中IL-6含量显著升高(P<0.05、P<0.01和P<0.001),细胞内脂质明显蓄积。150和300μmol/L油酸钠干预L02肝细胞TLR2表达显著高于对照组(P<0.05和P<0.001),TLR4表达未见显著上升。油酸钠各剂量组SIRT1蛋白表达低于对照组,NF-κB p65表达高于对照组。结论油酸钠引起L02肝细胞脂肪变性可能与TLR2/NF-κB介导的炎症通路有关。OBJECTIVE To observe the steatosis and inflammatory effects of L02 hepatocytes induced by different concentrations of sodium oleate.METHODS L02 cells were incubated with sodium oleate in different concentration for 24 h,and the cell viability was detected.L02 cells were respectively cultured with 75,150,300μmol/L sodium oleate for 24 h.The lipid accumulation of the cells was observed by oil red staining.The content of triglyceride in the cells was detected,the IL-6 content in the cell supernatant was detected.The expression of SIRT1 and nuclear factor-κB(NF-κB)was detected by Western Blot.The expression of Toll-like receptor 2 and Toll-like receptor 4(TLR2 and TLR4)on the cell surface was detected by flow cytometry.RESULTS With the increase of sodium oleate concentration,the cell viability decreased,the cell growth inhibition rate increased.The content of triglyceride in L02 cells treated with 75,150 and 300μmol/L sodium oleate was significantly higher than that in the control group(P<0.01,P<0.001,P<0.001),the IL-6 in the supernate was significantly higher than that in the control group(P<0.05,P<0.01,P<0.001).The result of oil red staining showed that the lipids in the cells were obviously accumulated after sodium oleate treatment.The expression of TLR2 in L02 cells treated with 150 and 300μmol/L sodium oleate was significantly higher than that in the control group(P<0.05,P<0.001).There was no significant increase in TLR4 expression after sodium oleate intervention in L02 cells.The expression of SIRT1 protein in the sodium oleate group was lower than that in the control group,and the expression of NF-κB p65 was higher than that in the control group through Western Blot result.CONCLUSION L02 hepatocyte steatosis caused by sodium oleate may be associated with TLR2/NF-κB mediated inflammatory pathway.

关 键 词：油酸钠 L02肝细胞 TOLL样受体2 核因子-ΚB 

分 类 号：R589.2[医药卫生—内分泌] Q593.1[医药卫生—内科学]