| Objective:1.Observe the clinical efficacy and safety of Peiyuan Dingchuan Decoction in treating patients with chronic persistent asthma(deficiency asthma syndrome),as well as its impact on serum IL-4,Arg-1,and CD163.2.By establishing an asthma mouse model and inducing M2 polarization of macrophages with IL-4,the effects of Peiyuan Dingchuan Decoction on airway inflammation and JAK1/STAT6 signaling pathway in asthma mice were observed,and the mechanism of action of Peiyuan Dingchuan Decoction on airway inflammation in asthma was explored.Methods:1.Clinical study:A total of 70 patients with chronic persistent asthma who were admitted to the Respiratory Department of the First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine from January 2021 to December 2022 and diagnosed with deficiency asthma were selected.They were randomly divided into a treatment group of 35 cases and a control group of 35 cases using a random number table method.The control group was given Xinbike Dubao,160μg/4.5μg/inhalation,twice daily;The treatment group was given Peiyuan Dingchuan Decoction twice a day on the basis of the control group,with a course of 4 weeks.Among them,one case in the control group was excluded due to not following the prescribed course of medication,and one case was excluded due to taking other drugs.33 cases were actually completed;One case in the treatment group was excluded due to failure to follow the prescribed course of medication,and 34 cases were actually completed.Before and after treatment,TCM syndrome scores,wheezing sounds,ACT scores,FeNO concentration,WBC,Neut,and EOS were compared between the two groups of patients,and changes in serum IL4,Arg-1,and CD163 levels were detected before and after treatment.2.Experimental study:40 C57BL/6 mice were randomly divided into 4 groups,with 10 mice in each group,namely the blank group,model group,budesonide group,and Peiyuan Dingchuan Decoction group.Except for the blank group,the model group,budesonide group,and Peiyuan Dingchuan Decoction group were used to establish asthma mouse models through ovalbumin(OVA)atomization and stimulation.Starting from the 16th day of modeling,the blank group and model group were given saline(0.1ml/10g)by gavage,the budesonide group was given budesonide(0.9%saline+lmg budesonide suspension)by nebulization for 30 minutes,and the Peiyuan Dingchuan Decoction group was given Peiyuan Dingchuan Decoction(18.75g/kg)by gavage,once a day for 4 consecutive weeks.After 6 weeks,observe the general state of each group of mice;WBC,EOS,and macrophage count in BALF;ELISA detection of IL-4,IL-5,IL-13 in BALF and IgE levels in serum;HE and PAS staining methods were used to observe the pathological changes in mouse lung tissue;flow cytometry was used to detect the expression of M1 and M2 macrophages in lung tissue;Western blot method was used to detect the expression level of IL-4 protein and the relative expression levels of p-JAK1/JAK1,p-STAT6/STAT6 proteins;the expression of IL-4,JAK1,STAT6,and Arg-1 mRNA in lung tissue was detected by qRT PCR method;immunohistochemical method was used to detect the expression of Arg-1 and CD163 in lung tissue;immunofluorescence staining was used to detect the expression of IL-4 in lung tissue.Results:Clinical research:1.Before treatment,there was no statistically significant difference between the treatment group and the control group in baseline data,TCM syndrome scores,wheezing sounds,FeNO concentration,ACT scores,WBC,Neut,EOS,and serum levels of IL-4,ARG-1,and CD163(P>0.05),which was comparable.2.In the treatment group,the controlled display rate was 54.54%,and the total effective rate was 93.94%;The control group had a controlled display rate of 35.29%and a total effective rate of 88.23%.There was no significant difference in the total effective rate between the two groups(P>0.05),but the treatment group had a better controlled display rate than the control group(P<0.05).3.After treatment,the total score of TCM syndromes in the treatment group and the control group were significantly lower than before treatment(P<0.05),and the treatment group was lower than the control group(P<0.05).4.After treatment,the individual scores of shortness of breath,shortness of breath,chills,cough,expectoration,soreness of waist and knees,and whiteness of face in both groups were significantly lower than those before treatment(P<0.05).5.After treatment,the score of wheezing sound in the treatment group and the control group was significantly lower than before treatment(P<0.05).After treatment,there was a statistically significant difference between the treatment group and the control group in wheezing sounds(P<0.05)6.After treatment,the ACT scores in the treatment group and the control group were significantly lower than before treatment(P<0.05).After treatment,there was a statistically significant difference in the ACT score between the treatment group and the control group(P<0.05)7.After treatment,WBC,Neut,and EOS in both groups were significantly lower than before treatment(P<0.05),and the treatment group was significantly lower than the control group(P<0.05).8.After treatment,the serum IL-4,Arg-1,and CD163 in the treatment and control groups were significantly lower than those before treatment(P<0.05).After treatment,the serum levels of IL-4,Arg-1,and CD163 in the treatment group were significantly different from those in the control group(P<0.05).9.The detection of safety indicators in both groups of patients before and after treatment has no clinical significance.Animal experiment:1.HE staining:In the blank group,there was no significant pathological change in the lung tissue HE staining;Compared with the blank group,HE staining in the lung tissue of mice in the model group showed a large number of inflammatory cells infiltrating,tracheal mucosa wrinkling,lumen narrowing,alveolar septa thinning and breaking,and adjacent alveoli fusing into larger cysts.Compared with the model group,the above pathological characteristics in the budesonide group and the Peiyuan Dingchuan Decoction group were significantly reduced.2.PAS staining:In the blank group,there was no obvious goblet cell proliferation and increased mucus secretion in the lung tissue PAS staining;Compared with the blank group,PAS staining in the lung tissue of the model group mice showed a significant increase in goblet cells and a large amount of airway mucus secretion;Compared with the model group,the proliferation of airway goblet cells and mucus secretion in the budesonide group and the Peiyuan Dingchuan Decoction group were significantly reduced.3.Compared with the blank group,the levels of IL-4,IL-5,IL-13 in BALF and IgE in serum of model group mice were significantly increased,while the number of WBC,EOS,and macrophages in BALF was significantly increased(P<0.05);Compared with the model group,both the budesonide group and the Peiyuan Dingchuan Decoction group decreased(P<0.05),and there was no difference between the two groups(P>0.05).4.Flow cytometry analysis showed:compared with the blank group,the percentage of macrophages of M1 type(F4/80+CD86+)and M2 type(F4/80+163+)in the model group was significantly higher(P<0.05);Compared with the model group,the percentage of Ml and M2 type macrophages in the budesonide group significantly decreased(P<0.05).5.Western blot showed:compared with the blank group,the expression of IL-4 protein and the relative expression of p-JAK1/JAK1,p-STAT6/STAT6 protein in the lung tissue of the model group mice were significantly increased(P<0.05);Compared with the model group,the protein expression levels in the budesonide group and the Peiyuan Dingchuan Decoction group were significantly lower(P<0.05);There was no difference between the budesonide group and the Peiyuan Dingchuan Decoction group(P>0.05).6.qRT-PCR detection showed:compared with the blank group,the expression of IL-4,JAK1,STAT6,and Arg-1 mRNA in the lung tissue of the model group mice was significantly decreased(P<0.05);Compared with the model group,the gene expression in both the budesonide group and the Peiyuan Dingchuan Decoction group was significantly lower(P<0.05);There was no difference between the budesonide group and the Peiyuan Dingchuan Decoction group(P>0.05).7.Immunohistochemical examination showed:the expression of Arg-1 and CD163 in the lung tissue of the model group mice was significantly higher than that of the blank group(P<0.05);Compared with the model group,budesonide group and Peiyuan Dingchuan Decoction group significantly decreased(P<0.05).There was no difference between the budesonide group and the Peiyuan Dingchuan Decoction group(P>0.05).8.Immunofluorescence staining showed:compared with the blank group,the fluorescence intensity of IL-4 in the model group was significantly enhanced;Compared with the model group,the fluorescence intensity of IL-4 in the budesonide group and the Peiyuan Dingchuan Decoction group was significantly reduced.Conclusions:1.Peiyuan Dingchuan Decoction can improve the traditional Chinese medicine syndrome efficacy,wheezing sound,ACT score,and FeNO concentration in patients with chronic persistent asthma(deficiency asthma syndrome),reduce the levels of WBC,Neut,EOS,and IL-4,Arg-1,and CD163 in serum,and safety.2.Peiyuan Dingchuan Decoction can alleviate the pathological changes of airway inflammatory cell infiltration,increased mucus secretion,and goblet cell proliferation in asthmatic mice,and reduce the level of Th2 type cytokines in asthmatic mice.3.Peiyuan Dingchuan Decoction can reduce the expression levels of Arg1 and CD163 in M2 macrophages mediated by IL-4.4.Peiyuan Dingchuan Decoction can reduce the expression levels of IL4,JAK1,and STAT6 proteins in the lung tissue of asthma mice.5.Peiyuan Dingchuan Decoction can inhibit macrophage M2 polarization by inhibiting the activation of IL-4/JAK1/STAT6 signaling pathway,thereby achieving the effect of treating asthma airway inflammation. |
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