SR-BI过表达对J774细胞胆固醇转运的影响

目的用构建的人SR-BI（hSR-BI）真核表达载体转染J774细胞,观察细胞瞬时过表达SR-BI对J774细胞胆固醇转运功能的影响。方法用hSR-BI cDNA转染法转染J774细胞,用RT-PCR和Western blot检测各组细胞的SR-BI表达,优化转染条件;用液体闪烁计数法检测流入和流出细胞的3H]胆固醇量,分析过表达SR-BIJ774细胞的胆固醇转运功能。结果与对照组和空载体转染的J774细胞相比,pcDNA3.1（-）＋hSR-BI转染的J774细胞SR-BI表达升高;与空载体转染的J774细胞相比,pcDNA3.1（-）＋hSR-BI转染的J774细胞的胆固醇双向转运功能增强。结论用hSR-BI真核表达载体pcDNA3.1（-）＋hSR-BI成功转染了J774细胞,使细胞过表达SR-BI,过表达的hSR-BI具有促进胆固醇转运的作用。

Effect of Overexpression of SR-BI on Cholesterol Trafficking in J774 Cells

Objective To investigate the effect of overexpression of SR-BI on the cholesterol trafficking in J774 cells. Methods J774 cells were transiently transfected by cloned hSR-BI to express hSR-BI, an 82 kDa protein. The expression of SR-BI were detected by RT-PCR and Western blot. The influx and efflux of 3 H ] cholesterol from cells were determined by liquid scintillation counting. Results The human scavenger receptor class B type I （hSR-BI） cDNA was successfully transfected into J774 cells. The expression level of SR-BI was elevated in J774 cells transfected with pcDNA3.1 （ - ） ＋ hSR-BI compared to control and vector （ P 〈 0.05 ）. The function of cholesterol trafficking in J774 cells transfected with pcDNA3.1 （ - ） ＋ hSR-BI was reinforced compared to control and vector. Conclusion Overexpression of SR-BI can promote cholesterol trafficking in J774 cells.