喘可治注射液对甲型H1N1流感病毒FM1株致小鼠肺炎的影响

Effect of Chuankezhi Injection on Prevention and Treatment of H1N1 Influenza Virus in Mice

目的:评价喘可治注射液对甲型H1N1流感病毒FM1株致小鼠肺炎模型的药效作用。方法:将ICR小鼠随机分为正常组、模型组、达菲组(27.5 mg·kg^(-1)·d^(-1))、喘可治注射液组(1.5 m L·kg^(-1)·d^(-1))。除正常组外,其余各组采用甲型H1N1流感病毒FM1株滴鼻感染ICR小鼠造成病毒性肺炎模型,分别进行死亡保护实验、治疗性给药实验和预防性给药实验。死亡保护实验中每组20只小鼠,以2倍半数致死量(LD_(50))流感病毒液滴鼻感染,感染后当天开始给药,每天1次,连续4 d,观察感染后14 d内动物的死亡情况,并计算死亡率、死亡保护率;治疗性给药实验中每组10只小鼠,以0.8倍LD_(50)流感病毒液滴鼻感染,感染后当天开始给药,每天1次,连续4 d,于实验第5天处死小鼠,采用酶联免疫吸附测定(ELISA)检测肺组织中白细胞介素-8(IL-8)含量及脑组织内发热介质前列腺素E_2(PGE_2),精氨酸加压素(AVP)的含量,实时荧光定量聚合酶链式反应(Real-time PCR)检测肺组织内相对病毒载量;预防性给药实验中每组10只小鼠,分组当天开始给药,每天1次,连续5 d,末次给药1 h后,除正常组外,其余各组以0.8倍LD_(50)流感病毒滴鼻感染ICR小鼠,感染后第4天称重后处死小鼠,计算各组肺指数、脾指数、胸腺指数及肺指数抑制率,Real-time PCR检测肺组织内相对病毒载量。结果:与正常组比较,模型组小鼠IL-8,PGE2含量,肺指数及肺组织内流感病毒株病毒载量均显著升高(P<0.01);与模型组比较,治疗性给予喘可治注射液可明显降低感染后小鼠的死亡率、肺组织内IL-8含量、脑组织内PGE_2含量及肺组织中流感病毒病毒载量(P<0.05,P<0.01);预防性给予喘可治注射液可显著提高感染小鼠的胸腺指数(P<0.01),明显降低感染小鼠肺指数及肺组织中流感病毒病毒载量(P<0.05);两种给药方式均可明显减轻支气管、血管周围及肺泡间质等组织炎性细胞浸润,减少管腔内红细胞渗出。结论:喘可治注射液对甲型H1N1流感病毒感染所致病毒性肺炎小鼠模型有较好的治疗和预防作用,其作用机制可能与减轻炎症反应和抑制病毒复制有关。

Objective: To evaluate the effect of Chuankezhi injection on mouse model of pneumonia induced by influenza A( H1N1) FM1 strain. Method: ICR mice were randomly divided into normal group,model group,tamiflu control group( 27. 5 mg·kg^-1·d^-1) and Chuankezhi injection group( 1. 5 m L·kg^-1·d^-1). In the death protection experiment,mice were infected with 2 × half lethal dose( LD50) of influenza virus FM1. The Chuankezhi injection was given once a day for 4 days. The number of death animal within 14 days was counted.The mortality and the death protection rate were calculated. In the treatment experiment,mice were infected with0. 8 × LD50 of influenza virus,and the Chuankezhi injection was given once a day for 4 days. On the 5thday after the infection,the levels of interleukin-8( IL-8) in lung,prostaglandin E2( PGE2) and vasopressin( AVP) in brain were tested by enzyme-linked immunosorbent assay( ELISA). The viral load of influenza virus in lung was tested by Real-time PCR. In the pre-treatment experiment,mice were given Chuankezhi injection once a day for 5 days. 1 hour after the last treatment,mice were infected with 0. 8 × LD50 influenza virus. 4 days after the infection,the lung index,spleen index,thymus index,and viral load in lung tissue were calculated. Result: Compared with normal group,the IL-8,PGE2 content,lung index and viral load in the lung tissue of model group were significantly increased( P < 0. 01). Compared with model group,Treatment of Chuankezhi injection could reduce the rate of deaths. Significantly inhibit the level of IL-8,PGE2,and the viral load of influenza( P < 0. 05,P <0. 01). Pre-treatment of Chuankezhi injection could significantly increase the thymus index of infected mice( P <0. 01),reduce lung index and the viral load of influenza( P < 0. 05). Both administration methods can significantly reduce the infiltration of inflammatory cells in the bronchial,perivascular and alveolar interstitium and reduce the exudation of red blood cells in the lumen. Conclusion: Chuankezhi injection could effectively prevent the mouse model of pneumonia induced by influenza A( H1N1) virus. The mechanism might be related to the reduction of inflammation and inhibiting viral replication.