18α-甘草次酸对斑马鱼急性毒性和对棕榈酸诱导的肝细胞脂质沉积的影响

Acute toxicity of 18α-glycyrrhetinic acid on zebrafish and effects of palmitic acid induced lipid deposition in hepatocytes

目的:基于模式生物斑马鱼,考察18α-甘草次酸(GA)的急性毒性。评价18α-GA对棕榈酸(PA)诱导的肝细胞脂质沉积的影响。方法:选取发育正常斑马鱼进行18α-GA急性毒性评价,绘制“浓度-死亡率”曲线,计算半数致死浓度(LC_(50))、10%致死浓度(LC_(10))和最大非致死浓度(MNLC)。考察不同浓度下,18α-GA对斑马鱼器官形态学结构畸形变化、运动情况异常变化和死亡数量等毒性表型的影响。采用PA诱导AML-12、HepG2及L02细胞,考察18α-GA对肝细胞脂质沉积的影响。结果:18α-GA对斑马鱼的LC_(50)、LC_(10)和MNLC分别为37、15、11μmol/L。一定浓度的18α-GA可诱发斑马鱼心脏、肝脏、肾脏毒性、躯干/尾/脊索弯曲以及肠道毒性。与PA(300μmol/L)组比较,18α-GA两浓度能够显著改善PA诱导的肝细胞胞质中脂质过量蓄积(P<0.05,P<0.01),10μmol/L的18α-GA能显著改善LDL、T-CHO及TG水平的异常升高(除HepG2细胞的T-CHO外)和HDL水平的异常降低(除HepG2细胞中HDL外)(P<0.01)。结论:斑马鱼急性毒性研究中,18α-GA具有潜在心脏、肝脏、肾脏和肠道组织毒性。18α-GA能够改善非酒精性脂肪肝病细胞模型脂质沉积。

Objective:To investigate the acute toxicity of 18α-glycyrrhetinic acid(GA)based on the model organism zebrafish.Meanwhile,the effect of 18α-GA on palmitic acid-induced lipid deposition in hepatocytes was evaluated.Methods:The normal developing zebrafish were selected to evaluate the acute toxicity of 18α-GA,and the‘concentration-mortality’curve was drawn to calculate the half lethal concentration(LC_(50)),10%lethal concentration(LC_(10))and maximum non-lethal concentration(MNLC)of zebrafish.The effects of 18α-GA on the morphological changes of zebrafish organs,abnormal changes in movement,number of deaths and other toxic phenotypes were investigated at different concentrations.AML-12,HepG2 and L02 cells were induced by palmitic acid(PA),and the effects of 18α-GA on the lipid deposition of hepatocytes were investigated.Results:The LC_(50),LC_(10) and MNLC of 18α-GA on zebrafish were 37,15 and 11μmol/L,respectively.Certain concentration of 18α-GA could induce cardiotoxicity,hepatotoxicity,nephrotoxicity,trunk curvature and enterotoxicity in zebrafish.Compared with the PA(300μmol/L)group,two concentration of 18α-GA could significantly reduce the PA-induced excess lipid accumulation in the cytoplasm of hepatocytes(P<0.05,P<0.01),10μmol/L 18α-GA could significantly improve the abnormal increase of LDL,T-CHO and TG levels(except the T-CHO in HepG2 cell)and the abnormal decrease of HDL level(except the HDL in HepG2 cell)(P<0.01).Conclusion:In the acute toxicity study of zebrafish,18α-GA may have potential hepatotoxicity,cardiotoxicity,nephrotoxicity and intestinal toxicity.18α-GA could improve lipid deposition in in vitro cell models of non-alcoholic fatty liver disease.