不同来源肝细胞在体外降脂药物评价中药效反应的对比

目的:通过对比不同来源的人肝癌细胞系HepG2和原代大鼠肝细胞在体外降脂药物评价中药效反应,指导两种肝细胞在体外降脂药物评价中的实际应用。方法:用游离脂肪酸(油酸/棕榈酸,2:1)诱导HepG2细胞、原代大鼠肝细胞脂肪变性,并用100μmol·L-1苯扎贝特干预,检测细胞内甘油三酯(TG)、总胆固醇(TC)、活性氧(ROS)含量,细胞内脂滴数目、并检测细胞上清液中丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性。结果:FFA刺激使HepG2细胞和原代大鼠肝细胞脂质沉积(TG、脂滴)和氧化应激(ROS、MDA、SOD)水平上升。苯扎贝特对HepG2细胞1 mmol·L-1FFA造模组和原代大鼠肝细胞0.5 mmol·L-1FFA造模组脂质沉积和氧化应激水平改善显著;而HepG2细胞0.5 mmol·L-1FFA造模组和原代大鼠肝细胞1 mmol·L-1FFA造模组脂质沉积和氧化应激水平在苯扎贝特干预后变化不明显。结论:在相同FFA造模浓度,原代大鼠肝细胞病理特征变化更为明显;苯扎贝特对两种肝细胞在脂质沉积和氧化应激水平的作用也不完全相同。因而HepG2细胞和原代大鼠肝细胞在体外降脂药物评价中药效反应是不完全相同的。

In-vitro Evaluation and Comparison of the Lipid-lowering Drugs Pharmacological Response on Liver Cells of Different Origin

Objective： To guide the application of the two kinds of liver cells in vitro evaluation of lipid-lowering drugs through evaluate the lipid lowering effects ofbezafibrate on liver cells from different origin with flee fatty acids model in-vitro. Methods： Hepatic steatosis was induced in HepG2 cells and primary rat hepatocytes by using free fatty acids （oleic acid / palmitic acid, 2：1） model. Lipid lowering effects of bezafibrate （100μmol. L-1） were analyzed by detecting various parameters： intracellular triglyceride （TG）, total chole- sterol （TC） and the lipid contents within cells. Further, Reactive oxygen Species （ROS）, malondialdehyde （MDA） and antioxidant enzymes （SOD） were examined as oxidative stress markers. Results： Hepatic steatosis （TG, lipid droplet） and Oxidative Stress （ROS, MDA, SOD） were increased with FFA treatment. Bezafibrate significantly ameliorate the hepatic steatosis and Oxidative Stress in HepG2 cells induced by 1 mmol- L-l FFA and primary rat hepatocytes induced by 0.5 mmol. L-1 FFA model. Howerever, bezafibrate have no significant amelio- rative effect on HepG2 cells induced by 0.5 mmol L-1 FFA and primary rat hepatocytes induced by 1 mmol. L-1 FFA model. Conclusions： In FFA induced hepatic steatosis model, disease pathological features are clearer in rat primary hepatocytes as compared to HepG2 cells. Moreover, pharmacological effects of bezafibrate are more significantly expressed at 1 retool. L-1 FFA induction in HepG2 and 0.5 mmol .L-1 FFA induction in rat primary hepatocytes. In-Conclusion pharmacological activity of lipid lowering drugs may differ in response when tested on HepG2 cells and primary rat hepatocytes at least in FFA model.