采用Changliver细胞制备肝细胞脂肪变性模型的方法探索

To explore the method of establishing hepatocellular steatosis models by Changliver cell

目的探索油酸(OA)诱导建立人正常肝细胞Changliver脂肪变性的离体细胞模型的实验条件。方法选用不同浓度油酸作用不同时间点诱导Changliver细胞,并用四甲基偶氮唑盐比色(MTT)法检测细胞活性,油红O染色观察细胞内脂滴数量、甘油-3-磷酸氧化酶法检测细胞内三酰甘油(TG)含量。结果采用0.2mmol/L OA诱导24h能建立较好的Changliver细胞脂肪变性模型,模型组细胞内TG水平为(379.98±23.19)mg/g,空白组细胞内TG水平为(185.03±12.68)mg/g,两组比较差异有统计意义(P<0.01)。结论 0.2mmol/L OA油酸诱导的Changliver细胞24h可建立比较稳定的脂肪变性模型。该模型为非酒精性脂肪性肝病的研究提供可靠的新途径。

Objective To explore the experimental condition for hepatocellular steatosis models of Changliver cell induced by o‐leic acid （lleic acid ,OA） .Methods Changliver cells were induced by different concentration of oleic acid for different periods .MTT was used to detect hepatic cell activity ,oil red 0 staining was used to observe intracellular lipid droplets acumulation ,glycerin 3 phosphate oxidase method was applied to detect the contents of triglyceride （TG） in the Changliver cell .Results Hepatocellular steatosis models of Changliver cell can be established successfully by 0 .2 mmol/L OA inducing for 24 hours .TG content in model cells was （379 .98 ± 23 .19）mg/g ,however ,it was （185 .03 ± 12 .68）mg/g in control cells ,the difference was statistically significant （P〈 0 .01） .Conclusion The proper condition for establishing hepatocellular steatosis models is 0 .2 mmol/L OA inducing Changliver cells for 24 h .This model is the reliable choice for nonalcoholic fatty liver disease research .