Qingkailing Injection On Experimental Studies Of Influenza Virus Infection In The Signal Transduction Pathways

Influenza A virus is the causative agent of the most common and highly infectious human disease.This cosmopolitan pathogen is responsible for a significant morbidity and mortality in humans as well as animals.Despite extensive clinical and pathological studies on influenza,our understanding of the mechanisms of disease development is still incomplete.The mammalian innate immune system recognizes the presence of invading pathogens by a family of receptors belonging to the Toll-like receptors(TLRs).Viral infection triggers an array of immune responses that effectively limit the replication and spread of viral infection.Recent studies have demonstrated that influenza viral infection up-regulates the expression of inflammatory cytokines,such as tumor necrosis factorα(TNF-α) mediated by NF-K B signal pathway.The host cleared the virus by the innate immune response,and trigger the subsequent specific immune response.If the pathway lost their normal control,the inflammatory hurt will be happened.The pathogenesis of influenza belongs to the theory "toxin hurts collaterals" of our Chinese medicine,and the toxin often result in the infectious lung hurt.The development of the influenza is process of the war between the inflammatory and anti-inflammatory.In this work,we investigated the antiviral ways and effect of the drug and the related signal pathway;We try to discover the antiviral molecular target of the Qingkailing injection combine with the Chinese medical theory.Objective:We try to find out the down -regulating effect by the NF-K B singal pathway of different drug-added Qingkailing Injection against influenza virus FM1 strain;At the same time Qingkailing Injection inhibite the expression of inflammatory cytokines, such as tumor necrosis factorα(TNF-α) in the mRNA level via the NF-K B singal pathway.we investigate the molecular mechanism that the drug inhibits the influenza virus so as to find the antiviral molecular targets of the Chinese medicine.Method:1.The luciferase assay result demonstrate the down -regulating effect by the NF- K B singal pathway of different drug-added Qingkailing Injection against influenza virus FM1 strain.2.Using RT-PCR detected the influence of the Qingkailing Injection on the antiviral signal transduction pathway mediated by the NF-K B in different concentration for 24 hours.To observe the change of the mRNA level of the inflammatory cytokine TNF-αResult:1.The luciferase assay result demonstrate that Qingkailing Injection can down regulate NF-K B singal pathway against influenza virus FM1 strain in dose-dependent manner.2.Qingkailing Injection can down-regulate the expression of mRNA of the signal proteins such asTNF-αin the signal transduction pathway mediated by the NF-K B.The down-regulation is significantly and shows dose dependent relationship.Conclusions:1.Qingkailing Injection can down regulate NF- K B singal pathway against influenza virus FM1 strain in dose-dependent manner.2.Qingkailing Injection can inhibite the expression of TNF-αin mRNA level and shows dose dependent relationship significantly through the NF-K B signal transduction pathway.