Study On The Effect Of Xifeng Dingchuan Recipe On Airway Inflammation In Asthmatic Guinea Pig Model Based On JAK2/STAT3 Signal Pathway

Objective:By replicating guinea pig model of cold asthma and using different doses of Xifengdingchuan recipe to intervene in guinea pig model of cold asthma,the effect of Xifengdingchuan recipe on airway inflammation of guinea pig and the influence on the expression of corresponding proteins and genes in the JAK2/STAT3 signaling pathway were observed,and the mechanism of Xifengdingchuan recipe on airway inflammation of guinea pig model of cold asthma was further studied.Methods:A total of 72 guinea pigs were randomly divided into normal group,model group,Xifengdingchuan recipe high-dose group(XFDCG group),Xifengdingchuan recipe medium-dose group(XFDCZ group),Xifengdingchuan recipe low-dose group(XFDCD group),and dexamethasone group(Dex group).After adaptive feeding for 7 days,guinea pigs in normal group were intraperitoneally injected on the 8th and 15 th day,respectively.The other groups were sensitized by injecting 1ml of 10% ovalbumin compound(ovalbumin compound was prepared by mixing ovalbumin and aluminum hydroxide with 0.9% sodium chloride solution at a ratio of 1∶100).From the 22 nd day after the experiment,except for the normal group,asthma was stimulated by ovalbumin(1%)atomization,30min/ time,once a day for 7 consecutive days,and cold stimulation was given for 1h after each end.The normal group was atomized with 0.9% sodium chloride solution and then placed in room temperature.After the successful establishment of cold asthma guinea pig model,intragastric administration began from the 29 th day.The equivalent dose of Xifengdingchuan recipe group and dexamethasone group was calculated according to the dose of children.The normal group and model group were given 0.9% sodium chloride solution(2.5ml/d)by intragastric administration for 21 consecutive days.During the modeling process,the general condition of guinea pigs was observed,and the body weight and death were recorded.At the end of intervention,samples were collected and the degree of inflammatory cell infiltration was observed by HE staining.The levels of IL-6,IL-17 and IL-23 in serum of guinea pigs with asthma were detected by ELISA.The protein levels of IL-6,IL-23 R,IL-17 R,JAK2,p-JAK2,STAT3 and p-STAT3 in guinea pigs with asthma were detected by Western Blot.Real-time RT-PCR was used to detect the mRNA expressions of IL-17 R,IL-23 R,JAK2 and STAT3 in the lung tissues of guinea pigs with asthma.Results:1.General observation: during the modeling period,compared with the normal group,guinea pigs in each group had less dry fur,lighter body weight,and decreased food intake with the increase of the number of excitation times.All guinea pigs gradually showed symptoms such as chilliness,decreased activity frequency,and slow reaction.After atomization stimulation,guinea pigs in the model group,Xifeng Dingchuan recipe group and dexamethasone group all showed symptoms such as irritability,sneezing and coughing,accelerated breathing,and even mouth breathing.After drug treatment,except for the model group and the normal group,the activity of guinea pigs in each group increased,the coat luster recovered and the amount of fur increased,the amount of food increased,the rate of weight growth accelerated,and the frequency of sneezing and coughing decreased.The improvement of general symptoms in the XFDCG group and the Dex group was better than that in the XFDCZ group and the XFDCD group.2.Pathological changes of lung tissue in guinea pigs in each group: In the normal group,the overall structure of the lung tissue was normal,the alveolar structure was clear,the alveolar wall was not significantly thickened,the alveolar collapse was not significant,and no obvious inflammatory cell infiltration was observed.Compared with the model group,Xifengdingchuan recipe dosage groups and western medicine groups showed mild pathological changes,no abnormal structural changes under lung tissue microscopy,low degree of airway wall hyperplasia,and no small amount of inflammatory secretions in the airway cavity.Among them,XFDCG group and DEX group showed the best improvement in pathological conditions under HE staining.3.Compared with the model group,the serum IL-6,IL-17,and IL-23 expression levels in the XFDCD group,XFDCZ group,XFDCG group,and Dex group were reduced.Among them,the IL-6 and IL-17 Dex groups decreased more significantly.IL-23 XFDCG group decreased more significantly(P <0.05);XFDCZ group,XFDCG group,Dex group lung tissue IL-23 R,STAT3,P-STAT3 protein levels decreased,of which the XFDCG group decreased more significantly(P <0.05);XFDCD Lung tissue IL-6,IL-17 R,JAK2,P-JAK2 protein levels in the XFDCZ group,XFDCZ group,XFDCG group,and Dex group decreased,among which the XFDCG group decreased significantly(P <0.05);The expressions of IL-17 R,IL-23 R,JAK2,and STAT3 mRNA in the lung tissues of the Dex group were all down-regulated.Among them,the IL-17 R mRNA XFDCG group decreased significantly,and the IL-23 R,JAK2,STAT3 mRNA Dex group decreased significantly(P <0.05).4.Compared with the normal group,the expression levels of serum IL-6 and IL-23 in model group,XFDCD group and XFDCZ group were increased(P < 0.05);Serum IL-17 and IL-17 expression levels in model group,XFDCD group,XFDCZ group,XFDCG group and Dex group were increased(P < 0.05).IL-23 R protein level in lung tissues of model group,XFDCD group,XFDCZ group and Dex group was significantly increased(P < 0.05);The protein levels of IL-6,IL-17 R,JAK2 and p-JAK2 in lung tissues of model group,XFDCD group and XFDCZ group were significantly increased(P < 0.05).The level of STAT3 protein in lung tissue of model group and XFDCD group was increased(P < 0.05).The level of p-STAT3 protein in lung tissues of model group,XFDCD group,XFDCZ group and Dex group was increased(P < 0.05).The mRNA expressions of IL-17 R,IL-23 R,JAK2 and STAT3 in lung tissues of model group,XFDCD group and XFDCZ group were up-regulated(P < 0.05).Conclusion:1.Xifeng Dingchuan recipe has an exact effect on guinea pig model of cold asthma.2.Xifeng Dingchuan recipe can significantly improve the inflammatory cell infiltration in guinea pigs with asthma airway.3.In this study,it was found that Xifengdingchuan formula could regulate the JAK2/STAT3 signaling pathway by regulating the expression of JAK2,STAT3,p-JAK2,p-STAT3 and other proteins,as well as interfering with the levels of inflammatory factors IL-6,IL-17,and IL-23 in serum,thereby reducing airway inflammation and playing an anti-inflammatory and antiasthmatic role.The mechanism may be related to the regulation of Th17/Treg cell imbalance.