The Comprehensive Evaluation On The Efficacy Kidney-yang Deficiency Mice Model Infected With Influenza Virus And The Intervention Effect On TLR3-TRIF-NF-κB Signaling Pathway Of Mahuang Xixin Fuzi Decoction

Objective: Previous studies have confirmed that the TLRs/NF- kappa B signaling pathway plays a innate antiviral immune response After influenza virus-infected cells,This study was to explore the need for The combination of syndrome with disease model,Using PLS to filter and evaluate the method of establish the kidney-yang deficiency mice model infected with influenza virus,To establish a stable and representative kidney-yang deficiency mice model infected with influenza virus.To analyze the pharmacodynamic mechanism and possible targets points of Mahuang Xixin Fuzi decoction in treating kidney-yang deficiency mice model infected with influenza virus from TLR3-TRIF-NF-k B signaling pathway,based on principle of "correspondence of prescriptions and syndromes".Methods: 1、Screening of kidney-yang deficiency mice model infected with influenza virus caused by different factors:Intraperitoneal injection of estradiol benzoate/hydrocortisone and then were infected with influenza virus H1N1/H3N2 to establish four kinds of kidney-yang deficiency mice model infected with influenza virus,using PLS comprehensive analysis of various indicators to evaluate and select the best model.2、To establish the kidney-yang deficiency mice model infected with influenza virus by the intraperitoneal injection of estradiol benzoate and infection with influenza virus H1N1, and intervene on the model with Mahuang Xixin Fuzi decoction. Analyzed the results by using partial least squares and evaluated the efficacy. 3、The m RNA and protein expression of influenza A H1N1, TLR3, TLR7, TRIF, My D88 and NF-KB(P50) in lung tissue of mice were detected by Western blot and RT-PCR technique, to analyze of the target points of Mahuang Xixin Fuzi decoction on the kidney-yang deficiency mice model infected with influenza virus.Results:1、Four kinds of the kidney-yang deficiency model mice appeared fur erection, dull, Listlessness, lethargy and other kidney-yang deficiency symptoms. Compared with normal control group,Anus temperature,Finger temperature and swimming time of the benzoate estradiol group were very significantly decreased(P < 0.01);toe temperature of the hydrocortisone group were significantly decreased(P < 0.05).The autonomic activitiesand swimming time of mice in the hydrocortisone group reduced,but there was no significant difference compared with normal control group.After virus infection, there was no death in normal control group, the model group mice had different degrees of death, the mortality rate of the estradiol benzoate H1N1 group reached 23.07%.Compared with the normal control group, the lung index of each model group was significantly increased and there was significant difference(P<0.05). 2、Body weight, Anus temperature, toe temperature, mortality, organ index and other indicators were analyzed by PLS. The data showed that each treatment group reflected a certain therapeutic effect on the kidney-yang deficiency mice model infected with influenza virus.The middle dose group of Mahuang Xixin Fuzi decoction showed better curative effect. 3、The experiments results of Western blot and RT-PCR show that compared with the normal control group,the m RNA expression of influenza A H1N1, TLR3, TLR7, NF-KB(P50) and the protein expression of TLR3, TRIF, My D88, NF-KB(P50) in lung tissue of the model mice Significantly increased(P<0.05).After drug intervention treatment,compared with the kidney-yang deficiency mice model infected with influenza virus group,the m RNA and protein expression of influenza A H1N1, TLR3, TLR7, TRIF, My D88 and NF-KB(P50) in lung tissue of mice in each treatment group showed a downward trend.Conclusion: 1、The results of the scoring chart and 2D drawing showed that the typical model,the kidney-yang deficiency mice model infected with influenza virus,could be established by useing this method of intraperitoneal injection of estradiol benzoate and infection with influenza virus H1N1. The Comprehensive efficacy score chart and the comparison chart showed that the middle dose group of Mahuang Xixin Fuzi decoction showed better curative effect. The middle dose of Mahuang Xixin Fuzi decoction is equivalent to the dose of clinical. This suggested that using Mahuang Xixin Fuzi decoction should pay attention to choose the appropriate dose range. Experimental results show that Pls can be simply and reliably used to analyze processing pharmacological data,can better reflect the characteristics of the syndrome and the overall regulation of the traditional Chinese Medicine and avoid of coming to a unilateral conclusion caused by evaluating one or several indicators in order to provide a more objective experimental basis for TCM clinical syndrome differentiation and disease differentiation. 2、The experiments of Western blot and RT-PCR proved that Mahuang Xixin Fuzi decoction could improve the signs of the model mice,Reduce lung pathological changes,improve the quality of life,down regulation the m RNA and protein expression of influenza A H1N1, TLR3, TLR7, TRIF, My D88 and NF- k B(P50) in lung tissue of mice and play a role of Innate anti viral immune response by inhibiting the activation of the TLR3-TRIF-NF- k B signaling pathway or reduced the expression of the key molecules.