Experimental Studies And Prevention Of Cangguo Sprays On Respiratory Viral Infections

Objective:Respiratory viral infection is a a clinical frequent disease and common disease of the threat to human health. Only some of the specific virus could be prevented by vaccine, but vaccine could not adapt to the virus mutation. The western medicine for respiratory viruses not only had strong side effects, but also easyly induced virus resistance. It will become a meaningful research to find effective drugs in Chinese medicine to create new pharmaceutical for the prevention of respiratory viral infections, based on the experience in prevention of epidemic diseases with Chinese medicine. Cangguo sprays was mucosal drug delivery formulations of traditional Chinese medicine(TCM), it was composed of borneol and the extracted volatile oil of the herb, caoguo, nepeta, elsholtzia, wrinkled giant hyssop, mint, cloves. Cangguo sprays was researched according to the theory of preventive treatment of disease and holistic concept and the principle of Fuzheng quxie in prevention of epidemic diseases in TCM, and combined with the by aromatic drugs, then it had the efficacy of dispelling wind and relieving, dispelling turbidness and dampness. In this study, the Asian influenza mouse lung to adapt to the strain (FM1) of influenza virus infected mice as a model, there were observed the effect of antiviral infected and the impact on immune function of Cangguo sprays, especially the impact of Cangguo sprays on non-specific immunity in the respiratory tract mucosal immunity, to explore its role in prevention of respiratory viral infections and immune regulation of the molecular mechanisms. And in vitro experiments there were carried out anti-influenza virus, respiratory syncytial virus, adenovirus, Coxsackie virus, and observed the role of prevention of common respiratory viral infections diseases of Cangguo sprays. Provide a theoretical basis for research and promote the use of Cangguo sprays.Methods:1. Cangguo sprays against four kinds of viral infections in vitroMonolayer cells inhibit anti-viral experiment werw researched by selecting for each virus-sensitive cells,, and observed the effect of anti-virus infection of Cangguo sprays on influenza virus A (H3N2), respiratory syncytial virus (RSV), intestinal Coxsackie virus (CVA16), adenovirus (Ad3).2. Cangguo sprays against FM1infected in vivo(1) lung index in FM1infected mice with Cangguo spraysKunming mice were randomly divided into seven groups:blank control group, model control group, positive control (ribavirin), Cangguo sprays solvent control group, Cangguo sprays hign, middle, low dose group, to15LD50influenza virus FM1virus liquid intranasal inhalation of infected mice model,50μl of everyone. Blank control group50μl normal saline intranasally. Modeling the first two days by the throat spray administration administration for consecutive seven days. The blank control group and model control group were given saline.24h after the last dose, mice were weighed and sacrificed, then removed the lung to calculate the lung index. Lung tissue were done HE dyeing to odserve lung pathological changes.(2) Immune function in FM1infected mice with Cangguo spraysAnimal grouping, modeling, administration method as above. The content of T lymphocyte subsets and NK cells in blood were detected by flow cytometry. By ELISA homogenate of mouse lung and mouse plasma were detected the pro-inflammatory cytokines of TNF-alpha, IFN-gamma, IL-2, IL-6, IL-8and the anti-inflammatory cytokines of IL-4and IFN-α content. The content of CD54and SIgA in the pharyngeal mucosal homogenates were determined by ELISA. Simultaneously the content of CD54and CD106in mucosa were detected immunohistochemical method.Results:1.Cangguo sprays against four kinds of viral infections in vitroIn the experimental results of resistance to the H3N2, RSV, CVA16and Ad3viruses in vitro, there were effectively improved the inhibitory rate of H3N2, RSV and CVA16virus with the different dosing of Cangguo sprays. There were inhibited virus adsorption and killed the H3N2, RSV, CVA16virus, then the replication and proliferation of the cells after virus adsorption of the H3N2, RSVand CVA16virus were inhibited with Cangguo sprays. There was no significant inhibitory effect on Ad3.2. Cangguo sprays against FM1infected in vivo(1) lung index in FM1infected mice with Cangguo spraysLung index of mice infected with influenza virus were significantly elevated. With Cangguo sprays there were effectively inhibited the increased lung index, and there were improved the pathological changes of lung tissue and were dose-dependented. Compared with the solvent control group there were significantly different (P<0.05).(2) Immune function in FM1infected mice with Cangguo spraysThe T lymphocyte subsets and NK cells in blood of influenza virus FM1mice were detected by Flow cytometry, CD3+, CD4+and CD4+/CD8+ratio were significantly decreased, and CD8+was significantly increased, and NK cells were significantly decreased. With each dose group of Cangguo sprays there were effectively increased CD3+, CD4+, CD4+/CD8+ratio and NK cell levels, and reduced CD8+content. By ELISA detection of the plasma of mice infected with influenza virus, the contents of IL-2, IL-4and IFN-a were significantly reduced and the contents of IL-6, IL-8were significantly increased. With each dose group of Cangguo sprays there were effectively increased the contents of IL-2, IL-4and IFN-a, and reduced the contents of IL-6, IL-8. By ELISA detection of lung homogenate of mice infected with influenza virus the contents of TNF-alpha and IFN-y were significantly elevated. With each dose group of Cangguo sprays the contents of TNF-alpha and IFN-y were effectively reduced. The contents of CD54and CD106of pharyngeal mucosa of mice infected with influenza virus with immunohistochemical detection were significantly increased, similarly the content of CD54of pharyngeal mucosal homogenate with the ELISA detection was increased, but the content of SIgA of pharyngeal mucosal homogenate with the ELISA detection was significantly reduced. With each dose group of Cangguo sprays, there were effectively reduced the contents of CD54and CD106, and effectively increased the content of SIgA.There were dose-dependent in each assay of Cangguo sprays. When each dose group of Cangguo sprays compared with the solvent control group, there were significantly different (P<0.05).Conclusion:1. In this paper, the theoretical research was according to the theoretical core of preventive treatment of disease, the theoretical basis of holistic concept and the basic principle of Fuzheng quxie in prevention of epidemic diseases in TCM. There were expounded the mechanism and advantages of prevention of respiratory virual infection with the external treatment of TCM, and there were elaborated the research overview of prevention of respiratory virual infection with TCM and western medicine. Then though experiment research and clinical prevent observation with Cangguo sprays, there were provided theoretical basis and experimental evidence for the clinical application of mucosal administration medicine of TCM for prevention of respiratory virus infections.2. There were effectively improved the inhibitory rate of H3N2, RSV and CVA16virus with the different dosing of Cangguo sprays in vitro. There were inhibited virus adsorption and killed the H3N2, RSV, CVA16virus, then the replication and proliferation of the cells after virus adsorption of the H3N2, RSVand CVA16virus were inhibited with Cangguo sprays. There was no significant inhibitory effect on Ad3.3. Cangguo sprays is an effective prescription for the prevention of respiratory viral infection. Lung index and lung tissue damage of FM1infected mice could be reduced with Cangguo sprays. It could be enhanced on non-specific immunity by increase the contents of NK cells and IFN-a with Cangguo sprays. Through the regulation of T cell subsets, reduce of pro-inflammatory cytokines and increase of anti-inflammatory factor, reduce of the inflammatory pathological damage, there were restored the stability and balance between cellular immunity and humoral immune function, then to enhance the ability of body to resist against respiratory viruses with Cangguo sprays.The expression of CD54and CD106in the pharyngeal mucosa could be reduced and the expression of SIgA in the pharyngeal mucosa could be increased with Cangguo sprays. By strengthening of the pharyngeal mucosa immunization against the virus function, and improving of neutralizing acttvity, and addition of the inhibition of virus attachment and invasion, there were enhanced the role of against respiratory viruses with Cangguo sprays.