甘草甜素对体外诱导的酒精性脂肪肝细胞的影响及机制研究
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摘要
目的探讨甘草甜素对体外诱导的酒精性脂肪肝(AFL)肝细胞模型的影响及其可能的作用机制。方法将体外诱导的AFL肝细胞分为对照组、脱离乙醇组、甘草甜素组和继续诱导组,检测细胞内三酰甘油(TG)水平,丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)的泄漏量、细胞周期变化,以及核受体超家族转录因子γ(PPAR-γ)、含有"碱性螺旋-环-螺旋-亮氨酸拉链"结构的核转录因子1(SREBP-1)、固醇调节元件结合蛋白裂解激活蛋白(SCAP)的mRNA与蛋白表达。结果甘草甜素可以明显减少细胞凋亡的发生;与对照组相比,甘草甜素组细胞内TG水平及ALT、AST的泄漏量、PPAR-γ、SREBP-1和SCAP蛋白及mRNA表达均明显降低(P<0.05)。结论甘草甜素可能通过下调PPAR-γ、SREBP-1、SCAP的表达从而减轻体外诱导AFL肝细胞脂肪改变。
Objective To investigate the effects of glycyrrhizin on the cell model of alcoholic fatty liver(AFL)induced in vitro and its possible mechanism.Methods The in vitro induced AFL cells were divided into the control group,separating ethanol group,glycyrrhizin group and continuous induction group.The levels of intracellular triglyceride(TG)and leakage amounts of ALT and AST,cell cycle change and protein and mRNA expressions of PPAR-γ,SREBP-1 and SCAP were measured.Results Glycyrrhizin could obviously reduce the apoptosis occurrence;compared with the control group,the level of intracellular TG and the leakage amounts of ALT and AST,and the protein and mRNA expressions of PPAR-γ,SREBP-1 and SCAP were significantly decreased(P<0.05).Conclusion Glycyrrhizin may alleviate the fat change of in vitro induced AFL cells by down-regulating expressions of PPAR-γ,SREBP-1 and SCAP.
Objective To investigate the effects of glycyrrhizin on the cell model of alcoholic fatty liver(AFL)induced in vitro and its possible mechanism.Methods The in vitro induced AFL cells were divided into the control group,separating ethanol group,glycyrrhizin group and continuous induction group.The levels of intracellular triglyceride(TG)and leakage amounts of ALT and AST,cell cycle change and protein and mRNA expressions of PPAR-γ,SREBP-1 and SCAP were measured.Results Glycyrrhizin could obviously reduce the apoptosis occurrence;compared with the control group,the level of intracellular TG and the leakage amounts of ALT and AST,and the protein and mRNA expressions of PPAR-γ,SREBP-1 and SCAP were significantly decreased(P<0.05).Conclusion Glycyrrhizin may alleviate the fat change of in vitro induced AFL cells by down-regulating expressions of PPAR-γ,SREBP-1 and SCAP.
引文
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[14]DA SILVA R P,KELLY K B,LEONARD K A,et al.Creatine reduces hepatic TG accumulation in hepatocytes by stimulating fatty acid oxidation[J].Biochim Biophys Acta,2014,1841(11):1639-1646.
[15]TOMIZAWA M,KAWANABE Y,SHINOZAKI F,et al.Elevated levels of alanine transaminase and triglycerides within normal limits are associated with fatty liver[J].Exp Ther Med,2014,8(3):759-762.
[16]SHI H,LUO J,ZHU J,et al.PPARγregulates genes involved in triacylglycerol synthesis and secretion in mammary gland epithelial cells of dairy goats[J].PPAR Res,2013,2013(6):310948.
[17]YOU M,JOGASURIA A,TAYLOR C,et al.Sirtuin 1signaling and alcoholic fatty liver disease[J].Hepatobiliary Surg Nutr,2015,4(2):88-100.
[2]PARK J H,LEE D H,PARK M S,et al.CCR5deficiency exacerbates alcoholic fatty liver disease through pro-inflammatory cytokines and chemokines-induced hepatic inflammation[J].J Gastroenterol Hepatol,2016,32(6):1258-1264.
[3]王宏英,陈林.甘草甜素抑制小鼠慢性支气管炎模型肺组织中炎性细胞因子表达的作用机制[J].中国实验诊断学,2016,20(3):372-374.
[4]李永伟,卢建溪,杨宏志,等.甘草甜素对HepG2.2.15细胞株CCL20的影响[J].中国实验方剂学杂志,2011,17(1):148-150.
[5]刘协,顾呈华,胡启之,等.甘草甜素脂质体对小鼠急性酒精性肝损伤的保护作用[J].中国临床康复,2004,8(36):8260-8261.
[6]张建,唐红敏,刘厚钰,等.甘草甜素多相脂质体对非酒精性脂肪肝肝功能的影响[J].肝脏,2003,8(3):30-32.
[7]马中春.甘草甜素对四氯化碳诱发大鼠肝脏损伤的作用及其机制[J].毒理学杂志,2005,19(3):250-251.
[8]李龙辉,李龙江,汤为学,等.甘利欣作用前后体外酒精性脂肪肝细胞模型甘油三酯水平变化及其可能作用机制[J].重庆医科大学学报,2010,35(2):179-182.
[9]张红锋,杨慧萍,王耀发.乙醇和软脂酸诱导的脂肪肝离体细胞模型[J].华东师范大学学报(自然科学版),2002(4):88-95.
[10]阮雪青.体外酒精性脂肪肝模型的建立和评价[D].杭州:浙江大学,2010.
[11]SHI D,ZHAN X,YU X,et al.Inhibiting CB1receptors improves lipogenesis in an in vitro non-alcoholic fatty liver disease model[J].Lipids Health Dis,2014,13(1):173.
[12]KAPIL S,DUSEJA A,SHARMA B K,et al.Genetic polymorphism in CD14gene,a co-receptor of TLR4associated with non-alcoholic fatty liver disease[J].World J Gastroenterol,2016,22(42):9346-9355.
[13]BOONVISUT S,NAKAYAMA K,MAKISHIMA S,et al.Replication analysis of genetic association of the NCAN-CILP2region with plasma lipid levels and non-alcoholic fatty liver disease in Asian and Pacific ethnic groups[J].Lipids Health Dis,2016,15(1):8.
[14]DA SILVA R P,KELLY K B,LEONARD K A,et al.Creatine reduces hepatic TG accumulation in hepatocytes by stimulating fatty acid oxidation[J].Biochim Biophys Acta,2014,1841(11):1639-1646.
[15]TOMIZAWA M,KAWANABE Y,SHINOZAKI F,et al.Elevated levels of alanine transaminase and triglycerides within normal limits are associated with fatty liver[J].Exp Ther Med,2014,8(3):759-762.
[16]SHI H,LUO J,ZHU J,et al.PPARγregulates genes involved in triacylglycerol synthesis and secretion in mammary gland epithelial cells of dairy goats[J].PPAR Res,2013,2013(6):310948.
[17]YOU M,JOGASURIA A,TAYLOR C,et al.Sirtuin 1signaling and alcoholic fatty liver disease[J].Hepatobiliary Surg Nutr,2015,4(2):88-100.