人血清白蛋白融合干扰素α2b体外抑制乙肝病毒的实验研究

Experimental study on inhibition of hepatitis B virus with human serum albumin interferon α2b in vitro

目的:研究长效干扰素人血清白蛋白融合干扰素(HSA-IFNα2b)对HepG2.2.15细胞增殖、凋亡及其HBsAg与HBeAg表达的影响,探讨其体外抑制乙肝病毒的机制。方法:采用SRB法分析HSA-IFNα2b作用HepG2.2.15细胞3,6 d后对其增殖的影响,酶联免疫分析不同剂量HSA-IFNα2b作用不同时间后HepG2.2.15细胞HBsAg与HBeAg的表达水平,Hochest33342、PI双染法观察细胞的凋亡和死亡,Western Blot印迹检测Bcl-2、Bax的表达。结果:HepG2.2.15细胞的增殖在加HSA-IFNα2b作用3,6 d后都有所抑制,细胞培养上清中HBsAg表达明显下降,并与浓度相关,HSA-IFNα2b浓度为10 000 U.mL-1时,培养6 d后HBsAg的抑制率达87%以上。Hochest33342、PI双染结果显示当HSA-IFNα2b浓度在1 250U.mL-1时可见少量的凋亡细胞,凋亡细胞数随着药物浓度的增大而增多,并有少量死亡细胞出现。Western Blot印迹检测随药物浓度的增加Bcl-2的表达降低而Bax的表达增加。结论:HSA-IFNα2b体外可明显抑制HepG2.2.15细胞的增殖及HB-sAg的表达,并可引起HepG2.2.15细胞的凋亡和死亡。

OBJECTIVE To evaluate the effects of a long-acting interferon （HSA-IFNa2b） on the proliferation, apoptosis and expression of HBsAg and HBeAg in HepG 2.2. 15 cells, and to investigate the mechanism of inhibition of hepatitis B virus in vitro. METHODS The cells proliferation was determined by SRB method. The expression levels of HBsAg and HBeAg in supernatant were measured using enzyme-linked dimmunosorbent assay （ELISA）. Apoptosis and death were observed by Hochest33342 and PI double staining. Expression of Bcl-2 and Bax proteins were determined with Western blot. RESULTS The results demonstrated that HSA-IFNa2b could inhibit the proliferation of HepG 2. 2. 15 cells. The expression of HBsAg in HepG 2. 2.15 cells was inhibited by HSA-IFNa2b in a time and dose-dependent manner. Treated with 10 000 u.mL^-1 HSAIFNa2b for six days, the inhibitiory rates of HBsAg was above 87%. The Results of Hochest33342 and PI double staining demonstrated HSA-IFNa2b could induce the apoptosis of HepG 2. 2. 15 cells in a dose-dependent manner. The cell death was also occurred at the higher concentration of HSA-IFNa2b. The results of Western blot analysis showed that HSA-IFNa2b could inhibit the expression of Bcl-2, while enhance the expression of Bax. CONCLUSION HSA-IFNa2b could not only significantly inhibit the HBsAg expression and the proliferation of HepG 2. 2. 15 cells, but also induced the apoptosis and death.