Study On Anti - Virus Effect And Mechanism

Objective:To investigate the anti-influenza virus of Ganduqing granule both in vivo and in vitro, and inspect the effect of Ganduqing granule on the induced interferon and Toll-like receptors signal pathway.Method:1. The survival rate of Ganduqing granule on the mice viral pneumonia model with influenza A/Sichuan/SWL/2009(H1N1) was inspected, the lung index, lung pathomorphological and pulmonary virus titer on the third day’s and sixth day’s were detected respectively, all that is to evaluate the anti-influenza virus effect in vivo of Ganduqing granule.2. To investigate anti-virus effect in vitro of ganduqing granule by means of detecting TCID50with cytopathic effect (CPE) of the MDCK infected model with influenza virus (A/Sichuan/SWL/2009(H1N1), A/Fujian Tong’an/196/2009(H3N2), B/Chongqing yuzhong/1384/2009, B/Sichuan anyue/139/2011) respectively.3. The influenza viral pneumonia model was built and the lung tissue was got on the third and sixth day with aseptic technique. The content of IFN-α、 IFN-β、IFN-γ in the lung homogenates was tested via ELSA method and the key targets, TLR3、TLR7、LLR8、TLR9、TRAF6、IRF3、IRF7、IRF8、c-Jun、NF-κB expression in Toll-like receptors signaling pathway were measured by Western Blot.Results:1. Ganduqing granule can enhance the survival rates, prolonged survival time, decreased or reversed the weight loss and disease incidence and severity of influenza viral pneumonic mice model. It decreased lung index, relieved pulmonary inflammation and fibrous degeneration on the third day’s and the sixth day’s and defered the course of desease, significantly reduced viral contents and virus duplicating speed in lung.2. The TCID50logarithm of influenza virus A/Sichuan/SWL/2009(H1N1), A/Fujian tong’an/196/2009(H3N2), B/Chongqing yuzhong/1384/2009, B/Sichuan anyue/139/2011affected MDCK were7.07,8.07,5.91,6.41respectively, the maximum non-toxic concentration of Ganduqing granule affected MDCK was530μg/ml. Ganduqing granule solution with maximum non-toxic concentration has no inhibitory action on the influenza virus A/Sichuan/SWL/2009(H1N1), A/Fujian tong’an/196/2009(H3N2), B/Chongqing yuzhong/1384/2009, B/Sichuan anyue/139/2011, and also no variance between the mouse serum which oral administered ganduqing granule and the control group.3. The protein level of TLR3, TLR7, TLR8were obviously higher than control group and were statistically significant (P<0.05). While there’s no statistically significant (P>0.05) with TLR9and IRF8. The protein levle of TRAF6, IRF3, IRF7, c-Jun and NF-κB p65were higher than control group and were statistically significant (P<0.05). It certified that influenza virus activated the Toll-like receptor signaling pasthway. Compared with the placebo, IFN-β of all the treated groups and IFN-α of Ganduqing granule’s middle dose group on the third day was significantly higher. There’s no statistical significance with IFN-γ. The protein level of TLR3, TLR7, TLR8, TRAF6, IRF3, c-Jun and NF-κB p65of Ganduqing granule groups were dose-dependent, obviously less than the placebo and were statistically significant (P<.05).Conclusion:Ganduqing granule showed conspiciously anti-virus effect in vito, but have no anti-virus effect in vitro, and the anti-virus effect is relating with inducing IFN-α, IFN/-β in early stage and antagonistating/inhibiting a few of proteins in Toll-like receptor signaling pathway.