The Effect Of Exterior-releasing Prescription On Nasal Mucosal Immune Barrier Dysfunction Of Mouse Induced By Cold Stimulation

Objective:This study investigated the effect and mechanism of Mahuang tang and Yinqiao powder which are the representative Exterior-releasing prescription on upper respiratory tract infection.The mouse nasal mucosal barrier dysfunction(MNMBD)model induced by cold stimulation was used to explore the effect of Mahuang tang and Yinqiao powder on improving mouse nasal mucosal barrier.We hope to uncover the scientific connotation of Exterior-releasing prescription on upper respiratory tract infection.Methods: 1.The effect of Exterior-releasing prescription on nasal mucosal immune barrier function in MNMBD model induced by cold stimulation 1.1 The effect of Mahuang tang on nasal mucosal immune barrier function in MNMBD model induced by cold stimulation Male KM mice were divided into Oseltamivir control group(ASTW+Normal),Mahuang tang control group(MH+Normal),normal group(Normal),model group(Model),Mahuang tang group(MH+Model)and positive drug group(ASTW+Model)randomly.Oseltamivir control group and positive drug group,Mahuang tang control group and Mahuang tang group were given oseltamivir phosphate(40mg/kg)and Mahuang tang(3.51g/kg)respectively for 2 days before modeling,once/day by intragastric administration;and we re-gave corresponding drugs 30 minutes before the model were induced.The other groups were given distilled water intragastrically.All animals were executed by excessive anesthesia to obtain the nasal mucosa tissues by conventional method at twenty-four hours after modeling.The protein expression of IgA and IFN-λ in nasal mucosa tissues of mouse was detected by Western-blot method.1.2 The effect of Yinqiao powder on nasal mucosal immune barrier function in MNMBD model induced by cold stimulation Male KM mice were divided into Oseltamivir control group(ASTW+Normal),Yinqiao powder control group(YQ+Normal),normal group(Normal),model group(Model),Yinqiao powder group(YQ+Model)and positive drug group(ASTW+Model)randomly.Oseltamivir control group and positive drug group,Yinqiao powder control group and Yinqiao powder group were given oseltamivir phosphate(40mg/kg)and Yinqiao powder(72.54g/kg)respectively 2 days before modeling,once a day by intragastric administration;and we gave corresponding drugs again 30 minutes before the model were induced.The other groups were given distilled water intragastrically.All animals were executed by excessive anesthesia to obtain the nasal mucosa tissues by conventional method at twenty-four hours after modeling.The protein expression of IgA and IFN-λ in nasal mucosa tissues of mouse was detected by Western-blot method.2.The mechanism of Exterior-releasing prescription on enhancing nasal mucosal immune barrier in MNMBD model induced by cold stimulation The epithelial cells produce IFN-λ,when it bind to the IFN-λ receptor,it can produce antiviral effector molecules such as myxovirus resistance protein 1(Mx1)to mediate the antiviral response of the innate immune system by activating the JAK1/ STAT1/2 signaling pathway.The protein expression of IL-10Rβ,IFNLR1,JAK1,STAT1,STAT2,p-JAK1,p-STAT1,p-STAT2,IRF9 and Mx1 were detected by western-blot method to observe the effect of Mahuang tang and Yinqiao powder on JAK1/ STAT1/2 signaling pathway.3.The effect of Mahuang tang on MNMBD model stimulated by Poly I:C Polyinosinic-polycytidylic acid(Poly I:C)is an artificially synthetic viral of double-stranded RNA analogue.Studies have reported that it can imitate virus invasion and cause upper respiratory tract infection.In order to investigate whether Mahuang tang can resist the damage caused by virus invasion,the Poly I:C was used to stimulate the model of MNMBD.3.1 The effect of Poly I:C stimulated on MNMBD model Male KM mice were divided into normal group(Normal),model group(Model),Poly I:C+Model group randomly.MNMBD model was made according to the previous method.The normal group and model group were stimulated with PBS through the nasal cavity at 0 hour after modeling.Poly I:C+Model group was stimulated with Poly I:C 5μg/μl,20μl per animal,through the nasal cavity at 0 hour after modeling.All animals were executed by excessive anesthesia to obtain the nasal mucosa tissues by conventional method at twenty-four hours after modeling.The protein expression of IgA and IFN-λ in nasal mucosa tissues of mouse was detected by Western-blot method.3.2 The effect of Mahuang tang on MNMBD model stimulated by Poly I:C Male KM mice were divided into normal group(Normal),Poly I:C+Model group,Mahuang tang group(MH+Poly I:C+Model)and positive drug group(ASTW+Poly I:C+Model)randomly.MH+Poly I:C+Model group and ASTW+Poly I:C+Model group were given Mahuang tang(3.51g/kg)and oseltamivir phosphate(40mg/kg)respectively by intragastric administration 2 days before modeling,once a day;we gave corresponding drugs again 30 minutes before the model were induced,the other groups were given with distilled water.Poly I:C stimulation were undergoing like the previous operation.All animals were executed by excessive anesthesia to obtain the nasal mucosa tissues by conventional method at twenty-four hours after modeling.Results: 1.The effect of Exterior-releasing prescription on nasal mucosal immune barrier function in MNMBD model induced by cold stimulation The results showed that Mahuang tang and Yinqiao powder could increase the protein expression of IgA and IFN-λ in mouse nasal mucosa of MNMBD model.2.The mechanism of Exterior-releasing prescription on enhancing nasal mucosal immune barrier in MNMBD model induced by cold stimulation The results showed that Mahuang tang could increase the protein expression of IFNLR1,p-JAK1,p-STAT1,p-STAT2,IRF9 and Mx1 on JAK1/ STAT1/2 signaling pathway(P(27)0.05 or P(27)0.01);Yinqiao powder could increase the protein expression of IL-10Rβ,IFNLR1,p-JAK1,p-STAT1,p-STAT2,IRF9,and Mx1(P(27)0.05 or P(27)0.01).3.The effect of Poly I:C stimulated on MNMBD model The results showed that the protein expression of IgA in the nasal mucosa of model group and Poly I:C+Model group were significantly reduced(P(27)0.01);the protein expression of IFN-λ in the nasal mucosa of the model group was lower than the normal group(P(27)0.05).However,the protein expression of IFN-λ in the Poly I:C+Model group was significantly higher vs.the model group(P(27)0.05).4.The effect of Mahuang tang on MNMBD model stimulated by Poly I:C The results showed that Mahuang tang increased the protein expression of IgA and IFN-λ in mouse nasal mucosa on Poly I:C stimulated MNMBD model(P(27)0.05 or P(27)0.01).Conclusions:Mahuang tang and Yinqiao powder could improve the immune barrier function of mouse nasal mucosa by increasing the protein expression of IgA and IFN-λ in nasal mucosal tissues.The mechanism of Mahuang tang and Yinqiao powder is mainly related to promoting the protein phosphorylation of JAK1 and STAT1/2,promoting the protein synthesis of IFN-λ,and increasing the protein expression of IFNLR1,IRF9,thereby promote the production of Mx1 which was an antiviral effector molecule.Mahuang tang could still improve the mouse nasal mucosal immune barrier function of MMNBD model with Poly I:C stimulation.