摘要
目的 探讨商陆抗病毒蛋白(PAP)对丙型肝炎病毒(HCV)感染细胞模型中HCV的抑制作用。方法 用HCV阳性血清感染人肝癌细胞株HepG2细胞。制成HCV感染细胞模型，用不同浓度的PAP干预该模型；利用荧光定量PCR法分别于药物干预后48h、96h、144h检测培养细胞及上清液中HCVRNA含量，同时以不同浓度干扰素(IFN)处理该模型作为对照。结果 PAP处理HepG2感染HCV细胞模型后。细胞内HCVRNA含量在48h、96h、144h各组问与对照组相比有显著差异(P<0．01)。在第48h，HCVRNA含量100μm／ml组和10μm／ml组<lμm／ml组<0．1μm／mll<0．01μm／ml组及对照组；在第96h，HCVRNA含量100μm／ml组和10μm／ml组<lμm／ml组<0．1μm／ml组和0．01μm／ml组<对照组；在第144h，HCVRNA含量100μm／ml组及10μm／ml组<1μm／ml组<0．1μm／ml组、0．01μm／ml组及对照组。培养上清液中HCVRNA含量各组问在48h时与对照组相比差异无显著性(P>0．05)，第96h，培养上清液中HCVRNA含量各组间差异显著(P<0．05)，100μm／ml组及10μm／ml组<1μm／ml组<0．1μm／ml组、0．01μm／ml组及对照组；第144h时。培养上清液中HCVRNA含量各组问差异显著(P<0．01)，100μm／ml组、10μm／ml组及1μm／ml组<0．1μm／ml组、0．01μm／ml组及对照组。PAP对HCV的抑制作用随PAP浓度的增加丽增强，以100μm／ml组对HCV的抑饲作用最强。IFN干预该模型亦得出相似结果，以3．0×10^4IU／ml组对HCV抑制作用最强。PAP与IFN均以第48h的抑饲效果最好，在48h、96h及144h时。PAP对HCV的抑饲作用显著高于IFN(P<0．01)。实验浓度的PAP未引起细胞死亡或脱壁，对细胞的形态、生长也无明显影响。结论 PAP对HCV复制有明显的抑制作用,且作用强于IFN。实验浓度的PAP对细胞形态及生长无明显影响。
Objective To investigate the inhibitory ef- fect of pokeweed antiviral protein (PAP) on the replication of HCV.Methods HCV RNA positive serum of a patient with HCV was used to infect human liver cancer cell line HepG2 to establish HCV infection cell model.Fluorescent quantified PCR assay was used to detect the level of HCV RNA in culturing cells and supernatant at 48h,96h,and 144h after the model was treated with various concentrations of PAP.Interferon (IFN) was used as a control.Results Compared with the con- trol,the level of HCV RNA in cells significantly decreased at 48h,96h and 144h after HepG2 model was treated with PAP. At 48h,the decrease of HCV RNA in various PAP concentra- tion group was following:100μtg/ml,10μtg/ml<1μg/ml group,1μg/ml<0.1μg/ml and 0.1μg/ml<0.01μg/ml and control.At 96h,100μg/ml,10μg/ml<1μg/ml;1μg/ml< 0.1μg/ml,0.01μg/ml;0.01μg/ml<control.At 144h, 100μg/ml,10μg/ml<1μg/ml;1μg/ml<0.1μg/ml,0.01μg/ ml and control. Compared with the control,the level of HCV RNA in su- pernatant did not significantly decreased at 48h after HepG2 model was treated with PAP.However,the decrease of HCV RNA at 96h and 144h had significant difference.At 96h,It was 100μg/ml,10μg/ml<1μg/ml<0.1μg/ml,0.01μg/ml and control,respectively.At 144h,it was 100μg/ml,10μg/ml and lμg/ml<0.1μg/ml,0.01μg/ml and control,respectively. The repression of PAP on HCV replication became strong when the concentration of PAP was high,and highest in 100μg/ml group.IFN was similar to PAP,and the inhibitory effect was the highest in 3.0×10~4IU/ml group.Both of PAP and IFN had strong inhibitory effect on HCV replication at 48h,while PAP had stronger repression than IFN.Conclusion PAP has significant inhibitory effect on HCV replication,and stronger than IFN.Experimental concentration of PAP had no influence on cell conformation and growth.
出处
《传染病信息》
2003年第3期134-134,共1页
Infectious Disease Information
