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目的通过体外实验研究β-谷甾醇与豆甾醇对非酒精性脂肪肝(NAFLD)发挥预防作用的量效关系。方法利用1.5μmol油酸处理LO2人肝细胞建立NAFLD细胞模型,药物组首先分别用10、20、50μmol/Lβ-谷甾醇或豆甾醇孵育细胞,之后用1.5μmol油酸处理LO2人肝细胞;实验检测细胞活力、甘油三酯(TG)、丙二醛(MDA)和谷胱甘肽(GSH)水平,并观察细胞形态的变化。结果总体上,β-谷甾醇或豆甾醇可显著降低脂肪肝细胞TG含量及脂质过氧化物MDA水平,提高抗氧化剂GSH含量,其中20μmol/L豆甾醇处理后细胞TG、MDA分别较模型组降低了66.6%和81.7%,而GSH含量增加了73.6%;10μmol/L和50μmol/Lβ-谷甾醇处理后,TG含量均比模型组低55.6%,MDA含量则分别降低了59.3%和62.9%,GSH含量分别增加了77.4%和72.6%。此外,两种植物甾醇对肝细胞增殖的抑制作用均随浓度升高而增强,但相同浓度豆甾醇对细胞活力的影响明显弱于β-谷甾醇。结论豆甾醇与β-谷甾醇可改善NAFLD肝细胞脂肪变性程度,减轻氧化应激反应,但作用效果呈不同量效关系。
Objective To study the dose-response relationship between β-sitosterol and stigmasterol in preventing non-alcoholic fatty liver disease (NAFLD) in vitro. Methods NAFLD cell model was established by treating LO2 human hepatocytes with oleic acid (1.5 μmol). The cells were incubated with 10, 20 and 50 μmol / L β-sitosterol or stigmasterol respectively, then treated with 1.5 μmol oleic acid for LO2 human hepatocytes. Cell viability, triglyceride (TG), malondialdehyde (MDA) and glutathione (GSH) levels were measured and the changes of cell morphology were observed. Results In general, β-sitosterol or stigmasterol significantly reduced the content of TG and lipid peroxidation in fatty liver cells, and increased the content of antioxidant GSH. The TG and MDA in 20μmol / L stigmasterol-treated group were significantly lower than those in model group The content of GSH was increased by 73.6%, the content of TG was 55.6% and the content of MDA was reduced by 59.3% and 62.9% respectively when treated with 10μmol / L and 50μmol / L β-sitosterol, , GSH content increased by 77.4% and 72.6% respectively. In addition, the inhibitory effects of both phytosterols on hepatocyte proliferation increased with increasing concentrations, but the effect of stigmasterol at the same concentration on cell viability was weaker than that of β-sitosterol. Conclusion Stigmasterol and β-sitosterol can improve hepatic steatosis and reduce oxidative stress in NAFLD patients, but the effect is dose-response relationship.