退热解毒灵水煎剂抗甲型H1N1流感病毒体内外实验研究

The Virostatic Action of Tuire Jieduling Decoction on H1N1 Type A Influenza Virus:In Vivo and in Vitro

目的观察退热解毒灵水煎剂对甲型H1N1流感病毒的抑制作用。方法体外抗病毒实验:将MDCK细胞悬液以105/ml接种于96孔板,用细胞维持液连续2倍系列稀释退热解毒灵水煎剂成5000、2500、1250、625、312.5、156.25、78、39、19.5mg/L9个稀释度,另将病毒唑注射液稀释成125、62.5、31.3、15.6、7.8、3.9、2.0、1.0、0.5mg/L9个稀释度,观察细胞病变效应,计算病毒抑制率。体内抗病毒实验:将80只BALB/c小鼠随机分为对照组、模型组、达菲组、退热解毒灵组,每组20只。除对照组外,其他各组小鼠均以10倍半数致死量剂量鼻腔接种甲型H1N1流感病毒50μl。感染当天开始给药,退热解毒灵组灌胃给予退热解毒灵水煎剂每天4133g/kg;达菲组灌胃给予达菲胶囊每天20mg/kg。每组取10只连续观察小鼠存活率;另10只于首次给药后4天、8天各处死5只,测定鼠肺湿质量及肺内病毒半数组织培养感染量(TCID50)。结果体外抗病毒实验:退热解毒灵水煎剂浓度为5000、2500、1250、625、312.5mg/L组和病毒唑注射液浓度为125、62.5、31.3、15.6mg/L组病毒抑制率均在50%以上。体内抗病毒实验:模型组小鼠存活率为0,达菲组与退热解毒灵组存活率分别为90%、60%。染毒后第4天、第8天,达菲组、退热解毒灵组肺湿质量及肺内病毒TCID50与模型组比较均明显下降(P<0.05或P<0.01)。结论退热解毒灵水煎剂对体内、体外甲型H1N1流感病毒均有一定的抑制作用。

Objective To observe the virostatic action of Tuire Jieduling Decoction（Decoction for effective defervescence and detoxification） on the H1N1 Type A influenza virus.Methods In vitro experiment：the MDCK cell suspension 105/ml was vaccinated onto the 96-hole plate,using the cell maintenance liquid to dilute continuously twice as much the Tuire Jieduling Decoction to 5000,2500,1250,625,312.5,156.25,78,39,and 19.5mg/L in concentration in series,and dilute twice as much the virazole injection to 125,62.5,31.3,15.6,7.8,3.9,2.0,1.0,and 0.5mg/L in concentration in series.The lesion effect of cells was observed and the virostatic rate was calculated.In vivo experiment：eighty BALB/c mice were randomized into control group,model group,Tamiflu group,and Tuire Jieduling group,with 20 in each.Except for the control group,mice of the other groups were vaccinated nasally with the H1N1 Type A influenza virus 50μl,a dose ten times of the half lethal dose（LD50）.The medication was given since the day of viral vaccination.Tuire Jieduling group was administered Tuire Jieduling Decoction 4133g/kg each day and the Tamiflu group was applied Tamiflu 20mg/kg each day through intragastric feeding.Ten mice of each group were observed on the survival rate,five were executed on the 4th day,and the other five on the 8th day after the first medication to measure the lung wet weight and intrapulmonary virus TCID50.Results In vitro experiment：the inhibiting rate of the 5000,2500,1250,625,and 312.5mg/L Tuire Jieduling Decoction subgroups and the 125,62.5,31.3,15.6mg/L virazole subgroups was all over 50%.In vivo experiment：the survival rate of the model group was 0,and that of the Tamiflu group and Tuire Jieduling Decoction group was 90% and 60% respectively.On the 4th day and 8th day after viral infection,the lung wet weight and intrapulmonary virus TCID50 of the Tamiflu group and Tuire Jieduling Decoction group were all significantly dropped in comparing with the control group,and the difference was statistically significant（P0.05 or P0.01）.Conclusion Tuire Jieduling Decoction is virostatic on the H1N1 Type A influenza virus both in vivo and in vitro.