Study On The Anti-influenza Virus Effect And Mechanism Of Schizonepeta Volatile Oil,Menthone And Pulegone

Objective:To study the anti-influenza virus activity of the volatile oil of Schizonepetae, menthone and pulegone and reveal its mechanisms by measuring the levels of cytokines relative to the acticity. Expecting to illustrate the scientific meaning of "relieving the exterior syndrome" and provide certain reference value for the interior discipline of "eliminating the exogenous pathogens" of warm-pungent diaphoretic drugs.Methods:The chick embryos and mice were infected by the mouse-adapted influenza virus A/PR/8/34strain, and to establish the mice viral pneumonia model.①Anti-influenza virus activity in chick embryos:the embryos were treated with different dosage of drugs, which were injected into the allantoic cavity and vitelline sac respectively in preventive and therapeutic administration way. Allantoic fluid was collected after incubating for72h, and the allantoic fluid HA on the chick embryo level was used as a measurement indicator.②Anti-influenza virus activity in mice: the mice which be infected with influenza virus were treated respectively with drugs in preventive and therapeutic administration way, and the death protection rate, lung index, lung tissue HA and pathomorphological change were adapted to evaluate the antiviral activity in mice.③The mechanism of anti-influenza virus:ELISA tests were conducted to determine the serum levels of IFN-a, IFN-β, IL-2, IL-6and TNF-a in virus infected mice.Results:①Allantoic fluid HA was significantly decreased after the chick embryos were treated with the volatile oil of Schizonepetae (0.05x2-15ml/egg) by the allantoic cavity injection in the therapeutic administration way, and with the volatile oil of Schizonepetae (0.1×2-12ml/egg,0.05×2-12ml/egg) by the vitelline sac injection both in the preventive and therapeutic administration way, meanwhile, with the volatile oil of Schizonepetae (0.025x2-12ml/egg) by the vitelline sac injection in the preventive administration way. In addition, allantoic fluid HA was significantly decreased after the chick embryos were treated with menthone (0.05×2-6ml/egg) by the vitelline sac injection in the therapeutic administration way, and with pulegone(0.1x2-8ml/egg) by the allantoic cavity injection in the preventive administration way and by the vitelline sac injection both in the preventive and therapeutic administration way, meanwhile, with the pulegone(0.05x2-8ml/egg,0.025×2-8ml/egg) by the vitelline sac injection both in the preventive and therapeutic administration way.②The volatile oil of Schizonepetae (0.452ml/kg,0.226ml/kg) has a significant effect on increasing death protection rate in preventive administration way, while the dosage of0.226ml/kg has a significant effect on increasing death protection rate, decreasing lung index and lung tissue HA, reducing the severity of pulmonary inflammation in therapeutic administration way. Methone(1ml/kg,0.5ml/kg) has a significant effect on increasing death protection rate in preventive administration way, while the dosage of0.5ml/kg has a significant effect on decreasing lung index and lung tissue HA in therapeutic administration way. In addition, Pulegone(0.19ml/kg) has a significant effect on increasing death protection rate in preventive administration way and decreasing lung index and lung tissue HA in therapeutic administration way.③The volatile oil of Schizonepetae(0.452ml/kg,0.226ml/kg) has a significant effect on Decreasing serum level of TNF-a in virus infected mice, while the dosage of0.226ml/kg has a significant effect on increasing serum levels of IFN-a、IL-2. Methone(0.5ml/kg) has a significant effect on increasing serum level of IFN-B and decreasing level of IL-6. In addition, pulegone (0.19ml/kg) has a significant effect on decreasing serum levels of IL-6and TNF-a.Conclusions:The experimental results showed that the volatile oil of Schizonepetae, menthone and pulegone have a good effect on anti-influenza virus both in vitro and in vivo, and the preventive administration way was inferior to therapeutic way. The mechanisms were related to the activity of increasing serum IFN-a, IFN-B, IL-2levels and decreasing IL-6,TNF-a levels which can improve immunoregulatory function and alleviate lung injury induced by influenza virus infection.