| Airway mucus hypersecretion is an important pathophysiological feature of acute exacerbation of chronic obstructive pulmonary disease(AECOPD).Previous studies had found that Qingjinhuatan Decoction has a good inhibitory effect to mucin 5AC(MUC5AC),the main component of mucus.In this paper,we began to explore the mechanism of Qingjinhuatan Decoction by animal experiments,and observe its clinical curative effect.This paper is divided into three parts:The first part is the literature review,introducing the relevant mechanism of immune regulation of airway mucus secretion in the acute exacerbation of chronic obstructive pulmonary disease and the progress of traditional Chinese medicine treatment of chronic obstructive pulmonary disease.The second part is experimental research.OBJECTIVE:To observe the effect of Qingjinhuatan decoction on immune regulation of airway mucus hypersecretion in acute exacerbation of chronic obstructive pulmonary diseaseMethods:The AECOPD-airway-mucus-hypersecretion rat models were built by the treatment of smoke and LPS airway injection.40 SPF WISTAR rats were randomly divided into blank control group,model group,Qingjinhuatan decoction group and clarithromycin group.From the first day of the experiment,the blank control group was fed normally without any intervention.The model group,Qingjinhuatan decoction group and clarithromycin group were separately treated with 0.9%sodium chloride solution,Qingjinhuatan Decoction,clarithromycin chloride Sodium solution.The drug doses are equivalent to the normal daily doses of adults,continuous gavage for 30 days.Observe the general situation of animals,record the weight of the rats.The lung function of the rats was measured on the 31st day,6 rats of each group were sacrficed after that.HE staining to observe the lung tissue,stain the goblet cells in the airway by alcian blue after iodine schiff.The expression of NE and MUC5AC in lung tissue was detected by immunohistochemical method.TLR4 mRNA,MyD88 mRNA and NF-κB mRNA were detected by real-time fluorescent quantitative PCR.The expression of TNF-α,IL-1β,IL-17,TGF-β and IL-10 in peripheral blood and alveolar lavage fluid were detected by ELISA.Flow cytometry was used to detect the expression of Treg and Thl 7 in peripheral blood.The expression of FOXP3 and ROR y t in lung tissue was determined by immunoblotting.Results:There were no significant differences in body weight among the four groups on the 1st and 10th day.On the 20th day,the body weight of the rats in the model group,Qingjinhuatan group and clarithromycin group were lower than those in the blank control group.(P<0.05).There was no significant difference among the model group,Qingjinhuatan group and clarithromycin group(P>0.05).On the 30th day,the weigt of the rats in the blank control group,the Qingjinhuatan group,and the clarithromycin group were higher than those in the model group(P<0.05).the weigt of the rats in the Qingjinhuatan group,and the clarithromycin group were lower than those in the blank control group(P<0.05).Compared with the blank control group,FEV0.3,FEV0.3/FVC%,MVV and PEF were downregulated in the model group,the difference was statistically significant(p<0.05,p<0.11,p<0.01,p<0.01).Compared with the model group,FVC,FEV0.3/FVC%and MVV were upregulated in the clarithromycin group,the difference was statistically significant(p<0.05,p<0.01,p<0.05).Compared with the model group,FEV0.3/FVC%and MVV of Qingjinhuatan group were upregulated,the difference was statistically significant(p<0.01).The number of goblet cells of the model group,the Qingjinhuatan group and the clarithromycin group were significantly higher than that of the blank control group,and the difference was statistically significant(p<0.05).Compared with the model group,The number of goblet cells of the Qingjinhuatan group and the clarithromycin group were significantly lower(p<0.01).There was no significant difference in the number of goblet cells between Qingjinhua sputum group and clarithromycin group(p>0.05).Immunohistochemistry showed that NE and MUC5AC were highly expressed in the airway.In the comparison of NE,the model group was significantly higher than that of the blank control group(P<0.01),the qinjinhuatan group was significantly higher than the blank control group(P<0.01)and significantly lower than the model group(P<0.05).There was no significant difference between Qingjinhuatan group and the clarithromycin group.In the comparison of Muc5AC,the model group,Qingjinhuatan group and clarithromycin group was significantly higher than the blank control group(P<0.01),qingjinhuatan group and the clarithromycin group were significantly lower than the model group(P<0.01,P<0.01),the Qingjinhuatan group was significantly lower than the clarithromycin group(P<0.01).Compared with the control group,the expression of TLR4,MyD88 and NF-κB in the model group were significantly increased,the difference was statistically significant(all p<0.01),the expression of TLR4,MyD88 and NF-κB in Qingjin Huatan group increased significantly(P<0.05,p<0.01,p<0.05).Compared with the model group,the expressions of TLR4,MyD88 and NF-κB in Qingjinhuatan group were significantly decreased,the difference was statistically significant(p<0.05,p<0.01,p<0.05)Compared with the control group,the levels of TNF-α and IL-1β in the serum and alveolar lavage fluid of the model group and Qingjin Huatan group were significantly increased(p<0.01).Compared with the model group,The levels of TNF-α and IL-1β in serum and alveolar lavage fluid of Jinhua sputum group were significantly decreased,the difference was statistically significant(p<0.05)Compared with the control group,the levels of IL-17 and IL-10 in serum and BALF were significantly decreased in the model group and Qingjin Huatan group(P<0.05,IL-10)0.01),the difference was statistically significant(serum IL-10 model group p<0.05,all were p<0.01);compared with the model group,Qinghua Huatan group serum and BALF IL-17 were significantly reduced,the difference was(P<0.01),but there was no significant difference between TGF-&and IL-10(P>0.05).The results of immunoblotting showed that there was no significant difference in Foxp3 expression between the three groups.There was significant difference between the three groups(P<0.01).Compared with the control group,the expression of ROR y t protein in the model group was significantly higher than that in the control group(P<0.01),and the expression of Foxp3/ROR y t was significantly lower than that of the control group(P<0.01)Significantly increased,the difference was statistically significant(all P<0.01).There was no significant difference in Treg ratio among the blank control group,model group and Qingjinuatan group.Compared with blank control group,Th17 ratio of model group upregulated significantly,Treg/Th17 downregulated significantly(P<0.05).There were no significant differences in Treg%,Th17%and Treg/Th17 between Qingjinuatan group and the model group.There was a significant negative correlation between MUC5AC and TGF-β,IL-10 in peripheral blood(P<0.01),and there was a significant positive correlation between RORγt,Th17,serum IL-17 and MUC5AC in lung tissue(P<0.01)Conclusion:1.Qingjinhuatan Decoction can upregulate FEV0.3/FVC%and MVV,inhibit the proliferation of airway goblet cells and downregulate the expression of NE and MUC5AC in lung tissue of AECOPD airway mucus hypersecretion model rats.2.Qingjinhuatan Decoction can downregulate the expression of TLR4 mRNA,MyD88 mRNA and NF-κB mRNA in lung tissue,and downregulated the levels of TNF-α and IL-1β in the serum and BALF of AECOPD airway mucus hypersecretion model rats.3,Qingjinhuatan Decoction can downregulate the expression of IL-17 in serum and BALF and RORγt in lung tissue of AECOPD airway mucus hypersecretion model rats.The third part is clinical research.OBJECTIVE:To observe the clinical effect of Qingjin Huatan Decoction on acute exacerbation of chronic obstructive pulmonary disease and to explore its effect on serum IL-17.Methods:Patients who meet the inclusion and exclusion criteria were randomly divided into two groups:the traditional Chinese medicine group and the control group.The patients in the control group were treated with routine western medicine.The patients in the traditional Chinese medicine group were treated with routine western medicine and Qingjinhuatan Decoction granules 1 bag bid,continuous treatment for 7 days.On 1st day and 7th day,took the venous blood measurement indicators and fill the CRF table.Observe:①general situation of age,gender,smoking history,past history②symptoms include chest tightness,wheezing,cough,sputum,irritability,fever,poor sleep,dry mouth,dry stool,yellow urine and so on,score 0-3 points from light to heavy quantification according to the patient’s subjective feelings,and calculate the total score.③mMRC score,CAT score(see attached table)④pulmonary function test FEV1/FVC,FEV1%expected ⑤C-reactive protein(CRP)⑥serum IL-17.According to the total score of the symptom before and after treatment,calculate the efficacy index for efficacy evaluation.Results:A total of 113 patients were collected and 10 cases were lost in the course of the reasearch.Among them,3 cases were from the control group and 7 cases were from the traditional Chinese medicine group.Two cases were removed from the control group.The data were completed in 101 cases,including 51 cases in the traditional Chinese medicine group and 50 cases in the control group.Before the treatment,the age,sex,CAT score,mMRC score,symptom score,CRP,severity of lung function,smoking history and serum IL-17 were compared between the two groups,there was no statistically significant difference(P>0.05).Both the control group and the traditional Chinese medicine group,the scores of the CAT score,mMRC,CRP and IL-17 significantly decreased after treatment(P<0.05).After treatment,the symptom scores of chest tightness,wheezing,cough,sputum,fever,yellow urine and total symptom scores of the patients of the control group were significantly lower than those before treatment(p<0.05).In terms of irritability,dryness,dry mouth,stool,there was no significant difference between before and after the treatment(p>0.05).The total symptom score and all the symptom except poor sleep significantly decreased(p<0.05).There was no significant difference between the two groups in terms of CAT score,mMRC score and CRP after treatment(p>0.05).Compared with the control group,the levels of IL-17 in the traditional Chinese medicine group were significantly lower than those in the control group(p<0.05).Compared with the control group,the symptoms and the total scores of cough,expectoration,irritability,dry mouth,dry mouth and dry stool were significantly lower than those of the control group(p<0.05).In term of chest tightness,wheezing,fever,yellow urine,poor sleep,there was no significant difference between the two groups(P>0.05).In the Chinese medicine group,clinical control 1,markedly effective 28,effective 21 ineffective 1,control rate was 56.86%,the total effective rate of 98.04%;In the control group,clinical control O,markedly effective 15,effective 26,ineffective 9,control rate 29.41%,total effective rate 80.39%.There were statistically significant differences between the two groups(p<0.01).Conclusion:1.Traditional western medicine on the basis of the addition of Qingjinuatan Tang,in improving the symptoms compared to the simple Western medicine group has a better effect,especially in the expectoration,irritability,dry mouth,dry stool and other symptoms.2.IL-17 may be the target of the treatment of AECOPD. |
