Experimental Research On The Effects Of Three Therapeutic Principles In Treating With Mice Infected By Influenza A (H3N2) Virus Under Different Environmental Conditions

The influenza virus is one of the serious pathogens which threat health of mankind, it is notoriously known for its unique ability to cause recurrent epidemics and global pandemics during which acute febrile respiratory illness explosively occurs among all age groups. In 1997, the affairs of human avian influenza occurred in Hong Kong. Continuously, someone was reported to be infected to death by highly pathogenic avian influenza virus (H5N1), the influenza vires demonstrated to mankind its huge "power" again.Influenza A (H3N2)vires is a subtype of the virus of influenza A. In 1968, its earliest stage was found in Hongkong, then disseminated to Japan, the United States etc. it caused the craze of flu in the world. Henceforth, although world craze did not happen again, but it has been popularizing up to now. In recent years, the world scope flu epidemiological supervision and investigating analysis manifested that influenza A (H3N2) virus is a popular advantage strain in Guangdong province, the whole country and all over the world.Modern investigation Of influenza A (H3N2) vires found that not only the international epidemic virus strain but also domestic epidemic virus strain cause antigenic drift. Owing to its importance to monitor the variation of H3N2 virus strain antigen to prevent flu from spreading and developing, so currently, abroad and local related organization have made a deep investigation to the antigenic variation of H3N2, especially from the molecularity of virus envelope top inset of Hemagglutinin (HA) and its head of the HA1 albumen code area of gene sequence, amino acids sequence and albumen molecularity structure were also carded on to clarify basically influenza A (H3N2) the molecular biology mechanism of the shift of virus antigen. But in treating with this subtype virus, in addition to the medicine of some anti influenza A virus, no new research progress have been made, and there is some medicine case report bearing the medicine of existing be continuously increasing. Influenza vaccines is the most valid means, but, because of the influenza virus continuously antigen vary, development of vaccine progress far lags variety speed of virus, so consumedly influence the effect of the vaccine, bring biggest difficulty to prevent from influenza.In Traditional Chinese Medicine, the syndrome of flu is just like exopathic disease. Its syndrome is included in not only wind-warm disease and spring-warm disease of epidemic febrile disease but also Taiyang disease, Yangming disease and Shaoyang disease of exogenous febrile diseases. The exopathic disease theory of TCM has long history and plentiful content, including a great deal of classical therapeutic principles and decoctions on treating flu. Nowadays, experimental research on anti influenza virus of TCM is spreading out actively, including research on antiviral herbs and decoctions in vitro and in vivo. These researches guide clinical treatment and can prevent of flu with TCM.At present, influenza A (H1N1) virus is made and used largely in experimental research on anti influenza virus of TCM, however, application of H3N2 is limited. At the same time, it is one of outstanding problem that syndrome differentiation and treatment formulation which is advantage of TCM can't be displayed adequately in current research which can't demonstrate clearly the exopathic disease theory, of TCM.Experimental research of our article is based on cloning and sequencing of HA gene of A/kunke/1/71 (H3N2) restrain. Research still includes cell culture in vitro and antiviral whole animal experiment in vivo. The aim of this experiment is to combine three therapeutic principles(releasing exterior with pungent-cool herbs, releasing exterior with pungent-warm herbs and releasing exterior with dispelling-dampness herbs) of TCM exopathic disease theory and three environmental conditions(normal environment, thermal environment and cold environment) to investigate the antiviral effects and immuno-regulation mechanism of compound prescriptions of TCM.The results on cloning and sequencing of HA gene indicate that max nucleotide homology is 99% between A/kunke/1/71 (H3N2) and A/Hong Kong/6/72 (H3N2) throng BLAST. Our research is the first time to perform HA molecular cloning and sequence analysis since A/kunke/1/71 (H3N2)restrain was clinically isolated in 1971. This is also the first time to identify this virus strain subtype on the level of gene.Experiment results in vitro indicate that representative decoctions of three therapeutic principles have no anti Influenza A (H3N2) virus functions directly on cell culture. Mechanism of the decoctions on anti influenza virus isn't the effects of exterminating or inhibiting influenza virus.Experiment of anti influenza virus in vivo results indicate three therapeutic principles have effects on treating mouse influenza A (H3N2) pneumonia in certain degree. They can reduce the score of lung consolidation (SLC), and relieve the pathomorphism seriousness of viral pneumonia. But there is variability on the effects of three therapeutic principles under different environmental conditions. Under normal and thermal environment, the principle of releasing exterior with pungent-cool herbs shows better antiviral effects , but it is ineffective under cold environment. Under different environmental conditions, the principle of releasing exterior with pungent-warm herbs and the principle of releasing exterior with dispelling-dampness herbs all show steadily satisfactory effects on anti influenza A (H3N2) virus in vivo.The results of analyzing correlative immune index indicate that three therapeutic principles have regulative effects on immune functions of mice infected by Influenza A (H3N2) vires. They can increase the percentage of mature T lymphocyte and T helper lymphocyte subsets (CD3~+ (%),CD4~+ (%)) in peripheral blood of infected mice. They also can promote the level of IL-2 and reduce the level of TNF-αin blood serum of infected mice. Accordingly, they can enhance cellular immune function and regulate balance between protective cytokine and pro-inflammatory cytokine. Moreover they may amend disorder of immune function and relieve immunological inflammation and pathologic injury which are caused by influenza virus. But it is different that three therapeutic principles have the regulative effects on immune function of mice infected under different environmental conditions. It is in thermal environment, the principle of releasing exterior with pungent-cool herbs shows better effects on enhancing cellular immune function and amending disorder of immune function. Only under normal environment and cold environment, the principle of releasing exterior with pungent-warm herbs just takes the effect on regulating the immunity. In different environmental conditions, the principle of releasing exterior with dispelling-dampness herbs performs steadily immunoregulational function.