寒喘祖帕颗粒对甲型流感病毒鼠肺适应株FM1感染肺炎小鼠的保护作用

Protective effect of Hanchuan Zupa granule on mice infected with influenza A virus FM1

目的研究寒喘祖帕颗粒对甲型流感病毒鼠肺适应株FM1感染小鼠的保护作用,并初步探讨其作用机制。方法①ICR小鼠60只,随机分为正常组、模型组、达菲组及寒喘祖帕颗粒高、中、低剂量组,每组10只。除正常组外,其余各组小鼠用甲型流感病毒鼠肺适应株FM1溶液(病毒原液稀释4.0×10~3倍)滴鼻感染小鼠造模。达菲组给予达菲47 mg/kg,寒喘祖帕颗粒高、中、低剂量组分别给予寒喘祖帕颗粒浸膏1.08 g/kg、0.54 g/kg、0.27 g/kg,正常组和模型组给予同体积蒸馏水。各组均连续干预5 d,于6 d时称取小鼠体质量,摘取全肺,计算其肺指数及抑制率;末次给药24 h后ELISA法测定小鼠血清肿瘤坏死因子(TNF-α)、γ干扰素(IFN-γ)水平;HE染色观察小鼠肺组织病理学改变并进行评分。②ICR小鼠160只,随机分为正常组、模型组、达菲组及寒喘祖帕颗粒高、中、低剂量组,正常组10只,其余各组每组30只。除正常组外,其余各组小鼠用流感病毒鼠肺适应株FM1溶液(病毒原液稀释3.5×10~3倍)滴鼻感染小鼠造模。造模当天开始各组分别灌胃给药,达菲组给予达菲47 mg/kg,寒喘祖帕颗粒高、中、低剂量组分别给予寒喘祖帕颗粒浸膏2.16 g/kg、1.08 g/kg、0.54 g/kg,正常组和模型组给予同体积的蒸馏水。各组均连续干预5 d,观察14 d内小鼠死亡情况,记录小鼠存活时间并计算死亡率。结果①与正常组比较,模型组小鼠体质量降低(P<0.01),肺指数增加(P<0.01),血清TNF-α水平升高(P<0.01),肺组织病理得分增加(P<0.01);与模型组比较,达菲组和寒喘祖帕颗粒高、中剂量组小鼠体质量增加(P<0.05,P<0.01),肺指数降低(P<0.05,P<0.01),血清TNF-α水平降低(P<0.01),肺组织病理得分降低(P<0.01);达菲组与寒喘祖帕颗粒各剂量组比较、寒喘祖帕颗粒各剂量组组间比较,上述指标差异均无统计学意义(P>0.05)。②与正常组比较,模型组小鼠存活时间减少(P<0.01),死亡率增加(P<0.01);与模型组比较,达菲组和寒喘祖帕颗粒高、中、低剂量组存活时间增加(P<0.01),达菲组寒喘祖帕颗粒高剂量组死亡率降低(P<0.01)。与达菲组比较,寒喘祖帕颗粒中、低剂量组存活时间减少(P<0.05),寒喘祖帕颗粒高、中、低剂量组死亡率升高(P<0.01)。与寒喘祖帕颗粒高剂量组比较,寒喘祖帕颗粒中、低剂量组存活时间减少、死亡率升高(P<0.05)。结论寒喘祖帕颗粒对甲型流感病毒鼠肺适应株FM1感染小鼠有一定的保护作用,该作用可能与抗炎、降低血清TNF-α含量有关。

Objective To study the protective effect of Hanchuan Zupa granule on mice infected by influenza A virus lung adaptive strain FM1,and to carry out a preliminary discussion of its mechanism.Methods①A total of 60 ICR mice were randomly divided into 6 groups:normal group,model group,Tamiflu group,and low,medium and high dose groups of Hanchuan Zupa granule,10 rats in each group.In addition to the normal group,mice in other groups were infected with nasal drops of influenza virus mouse lung adapted strain FM1 solution(virus original solution was diluted 4.0×10~3 times).Tamiflu group was given Tamiflu 47 mg/kg,and the high,medium and low dose groups of Hanchuan Zupa granule were given 1.08 g/kg,0.54 g/kg and 0.27 g/kg of Hanchuan Zupa granule extractum respectively.The normal group and the model group were given the same volume of distilled water.Each group was continuously intervened for 5 days,and the body weight of the mice was weighed on the 6 th day.The whole lung was extracted and the lung index and inhibition rate were calculated;the serum levels of tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ)were measured by ELISA 24 hours after the last administration;and pathological changes in lung tissue of mice were observed by HE staining and scored.(2)A total of160 ICR mice were randomly divided into 6 groups:normal group,model group,Tamiflu group,and low,medium and high dose groups of Hanchuan Zupa granule,10 rats in normal groups,and 30 rats each in other groups.In addition to the normal group,mice in the other groups were infected with nasal drops of influenza virus mouse lung adapted strain FM1 solution(virus original solution was diluted 3.5×103 times).Each group was administrated by gavage on the day of modeling,and the Tamiflu group was given 47 mg/kg Tamiflu,and the high,medium and low dose groups of Hanchuan Zupa granule were given 2.16 g/kg,1.08 g/kg and 0.54 g/kg of Hanchuan Zupa granule extractum respectively.The normal group and the model group were given the same volume of distilled water.Each group was continuously intervened for 5 days,and the death of the mice was observed within 14 days.The survival time of the mice was recorded and the mortality was calculated.Results(1)Compared with the normal group,the body weight of mice in the model group decreased(P<0.01),the lung index increased(P<0.01),serum TNF-αlevel increased(P<0.01),and the pathological score of lung tissue increased(P<0.01).Compared with the model group,the body weight of mice in the Tamiflu group and the high dose and medium dose groups of Hanchuan Zuopa granule increased(P<0.05,P<0.01),the lung index decreased(P<0.05,P<0.01),the serum TNF-αlevel decreased(P<0.01),and lung tissue pathological score decreased(P<0.01).There was no significant difference among Tamiflu group and Hanchuan Zuopa groups in the above indicators(P!0.05).(2)Compared with the normal group,the survival time of mice in the model group decreased(P<0.01)and the mortality increased(P<0.01).Compared with the model group,the survival time of mice in the Tamiflu group and the high dose,medium dose and low dose groups of Hanchuan Zupa granule increased(P<0.01),and the mortality of mice in the Tamiflu group and the high dose group of Hanchuan Zupa granule decreased(P<0.01).Compared with the Tamiflu group,the survival time of mice in the medium dose and low dose groups decreased(P<0.05),the mortality of mice in the high dose,medium dose and low dose groups of Huanchuan Zupa granule increased(P<0.01).Compared with the high dose group of Hanchuan Zupa granule,the survival time of mice in the medium dose and low dose groups decreased and the mortality of mice increased(P<0.05).Conclusion Hanchuan Zupa granule has certain protective effect on mice infected by influenza A virus lung adaptive strain FM1,and the effect may be related to anti-inflammation response and decrease of the TNF-αcontent in the serum of mice.