华支睾吸虫severin蛋白体外对HBV复制的影响

Effect of Clonorchis sinensis severin on HBV replication in vitro

目的探索华支睾吸虫severin(Cs severin)蛋白体外对乙肝患者外周血单核细胞(PBMC)乙肝病毒(HBV)复制水平的影响及对干扰素-α(IFN⁃α)清除HepG2.2.15细胞内HBV的影响。方法采用逆转录荧光定量PCR法(RT⁃qPCR)检测不同浓度(0、60、80、100μg/mL)Cs severin作用乙肝患者PBMC 48 h后HBV DNA拷贝数;采用RT⁃qPCR法检测不同浓度(0、60、80、100μg/mL)Cs severin作用HepG2.2.15细胞以及不同浓度(0、60、80、100μg/mL)Cs severin与IFN⁃α同时作用下的HBV DNA拷贝数;RT⁃qPCR检测HepG2.2.15细胞中STAT1 mRNA水平。结果Cs severin单独作用时不影响PBMC的HBV DNA拷贝,组间差异无统计学意义(P>0.05)。在HepG2.2.15细胞中,Cs severin的存在增强了IFN⁃α清除HBV的作用,且在60~100μg/mL范围内,随着Cs severin浓度的升高,HBV DNA拷贝数下降(P<0.05)。STAT1 mRNA水平在Cs severin存在的情况下高于对照组,各组间差异有统计学意义(P<0.05)。且随着Cs severin浓度的提高,STAT1 mRNA水平也随之上升。结论Cs severin不能直接影响HBV复制水平,但可能通过影响STAT1基因的表达而提高IFN⁃α清除HBV的能力。

Objective To explore the effect of Clonorchis sinensis severin(Cs severin)on hepatitis B virus(HBV)replication in peripheral blood mononuclear cell(PBMC)and HepG2.2.15 cells treated with interferon⁃α(IFN⁃α).Methods HBV DNA copies in the supernatant of PBMC were detected by reverse transcript real⁃time PCR(RT⁃qPCR)after incubation with different concentrations(0,60,80,100μg/mL)of severin for 48 h.HBV DNA copies in the supernatant of HepG2.2.15 cells were detected by RT⁃qPCR after administration with different concentrations(0,60,80,100μg/mL)of Cs severin with or without IFN⁃αfor 48 h.STAT1 mRNA level of HepG2.2.15 cells in each group was detected by RT⁃qPCR.Results There was no effect of Cs severin on HBV replication in PBMC(P>0.05).In HepG2.2.15 cells,the presence of Cs severin enhanced the effect of IFN⁃αon the elimination of HBV.In the range of 60⁃100μg/mL,with the increase of Cs severin concentration,HBV DNA copies decreased(P<0.05).In HepG2.2.15 cells,STAT1 mRNA levels were statistically higher in the presence of Cs severin than that in the control group(P<0.05).With the increase of Cs severin concentration,STAT1 mRNA levels also increased.Conclusion Cs severin could not directly affect the level of HBV replication but could improve the ability of IFN⁃αto eliminate HBV probably by affecting the expression of STAT1 gene.