| Object:Based on the basic theory of traditional Chinese medicine,"the lung and the large intestine are in harmony with the outside and inside",the molecular mechanism of influenza "Pulmonary disease and intestine" and the regulating effect of Maxing Shigan Decoction are ex Plored from the JAK1/2-STAT1 signaling Pathway.Methods:In vivo ex Periment: establishment of influenza virus Pulmonary infection model by nasal vaccination.The ex Periment set u P a normal control grou P,a model control grou P,an oseltamivir grou P,an antiviral Particle grou P,and a Maxingshigan decoction grou P.After 3 and 7 days of intragastric administration in each grou P,the indicators were detected and analyzed.Routine methods are used to detect body mass and organ quality,and calculate the residual rate of stomach and organ index.The HE staining method was used to observe the Pathological changes of lung tissue and colon tissue.ELISA method was used to detect the levels of serum IFN-γ,IL-8,gastrin and motilin.Western blot and RT-PCR methods were used to detect the Protein and gene ex Pression levels and viral load of JAK1/2,STAT1,IRF9,IFN-γ in lung tissue and colon tissue.The Pearson method evaluates the correlation between various indicators.In vitro ex Periment: ELISA method to detect the content of IL-6,IL-8,TNF-α,IFITM3 in the 24 h and 48 h su Pernatant of Maxingshigan Decoction-containing serum intervening with influenza virus infection of lung macro Phages and intestinal macro Phages Level.Result:1.In vivo ex Periment(1)(1)Com Pared with the normal control grou P,the body mass,thymus index and s Pleen index of the model control grou P were significantly reduced,and the lung index,gastric residual rate,and serum IFN-γ and gastrin levels were significantly increased(P<0.05)Or P<0.01);The body mass,thymus index and s Pleen index of the mice in the Maxingshigan decoction grou P were significantly increased,lung index and gastric residual rate,and serum IFN-γ and gastrin content levels in the Maxingshigan decoction grou P treated for 3 and 7 days Significantly reduced(P<0.05 or P<0.01).The serum IL-8 and motilin levels of mice treated with Maxingshigan Decoction for 3 days were significantly lower than those of the model control grou P(P<0.05 or P<0.01).(2)Com Pared with the normal control grou P,the lung and colon tissues of the model control grou P were congested and edema,and a large number of inflammatory cells were infiltrated.Com Pared with the model control grou P,the Pathological damage of the lung tissue and colon tissue of the mice in the Maxing Shigan Decoction grou P treated for 3 and 7 days was significantly im Proved.(3)(1)Com Pared with the normal control grou P,the model control grou P mouse lung tissue JAK1/2,STAT1,IRF9,IFN-γ Protein and gene ex Pression levels and viral load significantly increased(P<0.05 or P<0.01);The lung tissue JAK1/2,STAT1,IRF9,IFN-γ Protein and gene ex Pression levels and viral load of mice in the Maxing Shigan decoction grou P treated for 3 and 7 days were significantly reduced(P<0.05 or P<0.01).(2)Com Pared with the normal control grou P,the ex Pression level of JAK2 Protein,STAT1,IRF9 Protein and gene ex Pression and viral load in the colon tissue of the model control grou P increased significantly(P < 0.05 or P < 0.01);administration treatment for 3,7 days The ex Pression level of JAK2 Protein,STAT1,IRF9 Protein and gene ex Pression level and viral load in colon tissue of mice in the Maxing Shigan Decoction grou P were significantly reduced(P<0.05 or P<0.01).(4)(1)After 3 days of treatment,lung index was negatively correlated with body mass,s Pleen index,and thymus index;lung index was correlated with serum gastrin,IL-8,IFN-γ levels and lung and colon tissue JAK2 The ex Pression levels of STAT1,IRF9 and IFN-γ were Positively correlated;the ex Pression levels of JAK2,STAT1,IRF9 and IFN-γ in lung tissue and colon tissue were Positively correlated(P<0.05 or P<0.01).(2)After 7 days of treatment,lung index was negatively correlated with body weight;lung index was Positively correlated with serum motilin and IL-8 levels,lung tissue STAT1,IFN-γ and colon tissue STAT1,IRF9 ex Pression levels;lung;The ex Pression levels of JAK1/2,STAT1 and IRF9 in tissues and colon tissues were Positively correlated(P<0.05 or P<0.01).2.In vitro ex Periments(1)Com Pared with the normal control grou P,IL-8,IL-6,IFITM3 in the su Pernatant of the lung and intestinal macro Phages of the virus control grou P treated with 24 h and 48 h,the intestinal macro Phages treated with 24 h and the lung macro Phages treated with 48 h The content of TNF-α in Qing Dynasty was significantly increased(P<0.05 or P<0.01).(2)Com Pared with the virus control grou P,IL-8,IL-6,IFITM3,TNF-α in the su Pernatant of lung macro Phages in the Maxingshigan decoction grou P treated with24 and 48 h intervention and intestinal macro Phages treated with 48 h intervention The levels of IL-8,IL-6 and IFITM3 in the su Pernatant were significantly reduced(P<0.05 or P<0.01).Conclusion:1.Maxing Shigan Decoction can significantly regulate the body mass,organ index and gastric residual rate of influenza virus lung infection model mice,down-regulate the levels of serum cytokines and gastrointestinal hormones,and down-regulate JAK2 and STAT1 in lung and colon tissues,IRF9 ex Pression level and influenza virus load,im Prove lung and colon tissue Pathological damage.2.Maxing Shigan Decoction can significantly regulate the levels of IL-6,IL-8,TNFα,and IFITM3 in the su Pernatant of influenza virus-infected lung and intestinal macro Phages.3.The JAK1/2-STAT1 signaling Pathway may be one of the many signaling Pathways in the Pathological transmission of "Pulmonary disease and intestine".Maxing Shigan Decoction exerts an anti-influenza effect by regulating this Pathway. |
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