目的:研究泛素连接酶gp78在体外肝细胞脂肪变性发生过程中的作用。方法:在小鼠肝细胞内转染携带gp78的质粒DNA,400μM油酸刺激肝细胞,油红O染色观察细胞内脂滴形成的情况;测定肝细胞内甘油三酯含量;western blot和RT-PCR检测细胞内gp78的表达水平。结果:与对照组相比,过表达gp78肝细胞内脂滴数目增多,体积增大,甘油三酯含量(4.6±1.56)mol/L升高;而沉默gp78肝细胞内脂滴数目减少,体积减小,甘油三酯含量(0.3±1.37)mol/L明显降低(P〈0.05)。结论:在肝细胞发生脂肪变性过程中,gp78可能发挥了重要的作用;而其发挥作用的机制还有待进一步研究。
Objective: To study the role of gp78 in the process of fatty degeneration ofliver cells in vitro. Methods: Using oil red O staining to observe the formation of lipid droplets in mouse liver cells, determine triglyceride in liver cells. Expression levels ofgp78 in liver cells were detected by Western blot and RT-PCR. Results: As compared to the control group, Overexpression ofgp78 could increase the number of lipid droplets and upregulate triglyceride levels. While depletion of gp78 could decrease lipid droplets numbers, and also could downregulate the content of triglyceride. Conclusions: Our results strongly suggests that gp78 might play an important role during the course of hepatic steatosis.