大青龙汤颗粒剂体外抗甲型H1N1流感病毒实验研究

Research on Anti-influenza A H1N1 Virus in Vitro and Antipyretic Experiments of Major Decoction of Green Dragon Granule

目的:探讨大青龙汤颗粒剂体外抗甲型H1N1流感病毒的药效及对脂多糖所致大鼠发热的解热作用。方法:以甲型H1N1流感病毒感染狗肾细胞(MDCK),以细胞病变效应法(CPE)与噻唑蓝比色法(MTT)相结合,探讨大青龙汤颗粒剂对甲型H1N1流感病毒生物合成的抑制作用、对病毒吸附、穿入细胞的阻断作用及对病毒的直接杀伤作用。以脂多糖复制大鼠发热模型,分别测定大鼠不同时间点的肛温,绘制体温曲线,对解热作用进行分析。结果:大青龙颗粒剂对甲型H1N1流感病毒直接杀伤的半数有效浓度(IC50)为12.40 g·L-1,治疗指数(TI)为1.8。利巴韦林注射液的IC50为0.45 g·L-1,TI为1.0。模型组在造模后1 h即升温,5 h到达峰值,持续8 h。与模型组比较,大青龙汤颗粒剂高、低剂量组在造模后4～8 h有显著性差异。与阿司匹林组比较,大青龙汤颗粒剂高、低剂量组在1～3 h降温作用稍弱,第4小时、第5小时即与之相当,而第6小时至第8小时优于阿司匹林。大青龙汤颗粒剂高、低剂量组间无显著性差异,但低剂量组效果稍优于高剂量组。结论:大青龙颗粒剂在细胞水平表现出一定的抗甲型H1N1流感病毒作用,作用形式为直接灭活,且呈剂量依赖性;但其抑制流感病毒生物合成及阻断吸附、穿入细胞的作用不明显。其对脂多糖所致大鼠发热的解热作用显著,且药效持续时间长。

Objective： To discuss the effect of Major Decoction of Green Dragon Granule in anti-influenza A H1N1 virus in Vitro and its antipyretic influences to fever rats caused by lipopolysaccharide.Methods： MDCK infected by anti-influenza A H1N1 virus,and combined cytopathic effect（CPE） with MTT colorimetric assay,and the drug effect of Major Decoction of Green Dragon Granule inhibiting the biosynthesis of A H1N1 influenza virus,the absorption to influenza virus,the blocking of the accession to cells and killing influenza virus were inspected.Lipopolysaccharide duplicated fever models of rats were used,and rectal temperatures of rats of different groups in different times were measured,curves were drawn,and antipyretic effects were analyzed.Results： The half maximal inhibitory concentration（IC50） of Major Decoction of Green Dragon Granule killing influenza AH1N1 was 12.40 mg·mL^-1,the therapeutic index（TI） was 1.8.The IC50 of Ribavirin Injection in the same administration route was 0.45 mg·mL^-1 and TI was 1.0.The temperature of the model group had already rised at 1 h after establishing models,reached peak at 5 h,and sustained for 8 h.Compared to the model group,the Major Decoction of Green Dragon Granule of high-dose group and low-dose group had significant differences at 4-8 hour.Compared to aspirin group,the Major Decoction of Green Dragon Granule of high-dose group and low-dose group,the range of cooling the temperature was smaller at 1-3 hours,and was equal at 4-5 hour,but was larger than that of aspirin group at 6-8 hour.The difference between Major Decoction of Green Dragon Granule of high-dose group and low-dose group was not significant.Conclusion： Major Decoction of Green Dragon Granule shows certain effect of anti-influenza A H1N1 virus in cell level,and the function form is direct inactivation in a dose dependent manner;but the effect of Major Decoction of Green Dragon Granule in inhibiting the biosynthesis of influenza virus,absorbing to influenza virus,and blocking the accession to cells are not significant.Major Decoction of Green Dragon Granule has a significant antipyresis effect of fever rats caused by lipopolysaccharide,and the drug duration is longer.