| ObjectiveObservation for the assessment of strengthening earth to generate metal decoction in the treatment of chronic obstructive pulmonary disease patients in Stable phase with both lung and kidney deficiency syndrome of the pulmonary function,times and degree of acute exacerbation,TCM syndrome score,quality of life and 6MWD.To establish a rat model of chronic obstructive pulmonary disease(COPD),set up the normal group,COPD model group and the high,middle and low dose group,and collect the peripheral blood,bronchoalveolar lavage fluid(BALF)and lung tissue samples.Lung tissue was taken for paraffin section and HE staining.Flow cytometry was used to detect the changes of Th17 and Treg in peripheral blood,BALF and lung tissues.IL-17 and IL-23 were detected by ELISA method.To explore the method of reinforcing earth to generate metal through the Th17/Treg pathway,regulating the differentiation mechanism of balance and control in COPD.In order to further scientific evaluation of strengthening earth to generate metal decoction combined treatment on the clinical effects of COPD in stable phase.Methods1.Clinical observation:This test will meet the inclusion criteria of COPD stable lung spleen deficiency syndrome were randomly divided into strengthening earth to generate metal decoction Combined Treatment Chinese medicine treatment group and Western medicine control group.According to the control group of patients treated with routine treatment of Western medicine,Chinese medicine strengthening earth to generate metal decoction combined treatment group in the control group treated with conventional western medicine treatment of traditional Chinese medicine oral strengthening earth to generate metal decoction combined treatment for 3 months,to maintain the routine treatment after 3 months,whole treatment was 6 months.The number and degree of acute exacerbation,TCM syndrome score,CCQ questionnaire and six minute walking distance,plasma IL-17 and IL-23 were observed before treatment,3 months after treatment and 6 months after treatment.2.Experimental study:2.1 In this experiment,the use of cigarettes smoked to establish a rat model of chronic obstructive pulmonary disease lipopolysaccharide combined modeling method of filling tube,from the general state of rats was observed,lung tissue,lung tissue pathological observation,bronchoalveolar lavage fluid cytology evaluation of chronic obstructive pulmonary disease rat model,and the effect of traditional Chinese Medicine strengthening earth to generate metal method the rat pathological changes and inflammatory cells of chronic obstructive pulmonary disease.2.2 At sixteenth weeks,invasive lung function tests were performed in all groups:PIF(inspiratory peak flow rate),PEF(expiratory peak flow rate),VC(vital capacity),Cdyn(lung dynamic compliance),FRC(forced vital capacity)and MV(minute ventilation volume).To evaluate the rat model of chronic obstructive pulmonary disease and to evaluate the rat model of chronic obstructive pulmonary disease and hilling in traditional Chinese medicine health gold Decoction on chronic obstructive pulmonary disease rat airway resistance and lung compliance,to explore the reliability of experimental animal model.2.3 Adsorption method for determination of serum and bronchoalveolar lavage fluid of rats were detected by IL-17,the content of IL-23 by ELISA,explore the anti-inflammatory effects of strengthening earth to generate metal decoction combined treatment prescription on chronic obstructive pulmonary disease in rats.2.4 The expression of Th17 and Treg flow cytometry in rat peripheral blood,BALF and lung tissues,to explore the regulatory role of strengthening earth to generate metal decoction combined treatment prescription of chronic obstructive pulmonary disease in rats.Results1.Clinical research:1.1 By SPSS 21 statistical analysis,the two groups of patients with general information including age,gender,course of disease and severity of the disease were no significant difference(P>0.05),comparable.1.2 Effect of TCM syndromes:integral and total change trend of the syndrome from the observation line chart,strengthening earth to generate metal decoction group in the improvement of cough,expectoration,fatigue,abnormal stools of patients,the total score of TCM symptoms,and better than the western medicine control group(P<0.05),in the breath,self sweat,easy cold,anorexia side comparison and the control group had no significant difference(P>0.05).The two groups after 3 months of treatment of TCM symptoms comparison,strengthening earth to generate metal decoction combined treatment in Chinese medicine group in reducing phlegm,spontaneous sweating,abnormal defecation,cough,fatigue and other symptoms,total score and Western medicine contrast group,the difference was statistically significant(P<0.05),but no significant wheezing,colds,anorexia and other symptoms with the score of the western medicine group differences(P>0.05).The two groups after 6 months of treatment of TCM syndrome compared strengthening earth to generate metal decoction Combined Treatment Chinese medicine group decreased in anorexia,shortness of breath,cough,fatigue,abnormal stool score and total score compared with western medicine group,the difference was statistically significant(P<0.05),and phlegm,wheezing,easy cold difference score and Western medicine contrast group no statistical significance(P>0.05).1.3 The number and severity of acute exacerbation effect:the study during 6 months of observation,strengthening earth to generate metal decoction group and Western medicine control group in the times of acute exacerbation and acute exacerbation of the degree of contrast decrease,but the differences were not statistically significant(P>0.05).1.4 The influence of the quality of life:the indicators of quality of life before and after the study were compared with CCQ score,symptoms,functional status,mental status scores over time were decreased,but the Peitu Shengjin group and the control group had no significant difference(P>0.05).Before treatment and 3 months of treatment,two groups of CCQ score,symptoms,functional status,mental state of the difference was not statistically significant,and 6 months after treatment CCQ score,symptom score,mental state Peitu Shengjin medicine group was lower than that of control group(P<0.05),suggesting that.Native gold method can improve the patient’s quality of life,delay the decline in the quality of survival of patients with COPD.1.5 The effect of six minutes walking distance:strengthening earth to generate metal decoction group and Western medicine control group two group 6 minutes walking distance trends over time are roughly the same,with the passage of time increased,strengthening earth to generate metal decoction group increased more significantly,but the difference was not statistically significant(P>0.05).Between the two groups at different time points compared before treatment and 3 months after treatment,the two groups had no statistically significant difference between the 6 minute walk distance(P>0.05),and 6 months of treatment at 6 minutes walking distance from the traditional Chinese medicine group was higher than the control group(P<0.05),suggesting that strengthening earth to generate metal decoction combined treatment can improve tolerance ability and activity of active patients.1.6 Effect of peripheral blood IL-17 and IL-23:strengthening earth to generate metal decoction group and Western medicine control group,two groups of plasma IL-17 concentration trends over time are roughly the same,with the passage of time decreased,strengthening earth to generate metal decoction group decreased significantly compared with the control group(P<0.05).Between the two groups at different time points compared before treatment and 3 months after treatment,there was no significant difference in plasma IL-17 of the two groups(P>0.05),and 6 months of treatment by plasma IL-17 gold in Chinese medicine group was lower than control group,the difference was statistically significant(P<0.05).Strengthening earth to generate metal decoction group and Western medicine control group,two groups of plasma IL-23 concentration trends over time are roughly the same,with the passage of time decreased,but the difference was not statistically significant(P>0.05).Between the two groups at different time points compared before treatment and 3 months after treatment,there was no significant difference in plasma IL-23 of the two groups(P>0.05),and 6 months of treatment by plasma IL-23 gold in Chinese medicine group was lower than control group,the difference was statistically significant(P<0.05).2.Experimental study:2.1 Effect of strengthening earth to generate metal decoction on COPD rat weight and lung tissues,pathologyThe rats in the normal group showed a healthy state and maintained a steady and steady rise in weight.By way of LPS+fumigation model rats,rats appeared obvious early symptoms of anorexia,yellow hair gradually lost luster,listlessness,activity significantly reduced,some rats appeared sneezing symptoms after 4 weeks,most of the rats began to recover gradually,eating and drinking began to increase the amount of mental state began improved activity began to increase,but compared with the normal group rats is low activity.From the change of body weight of rats,4 weeks before modeling rats began in smoking,weight growth rate decreased significantly,model group,Chinese medicine group were significantly lower than the normal group,the difference was statistically significant(P<0.05),high dose of Chinese medicine group from the beginning of the eighth week weight growth rate compared with the model group speed up the trend(P<0.05).At twelfth weeks,the growth rate of body weight in the high,medium and low dose groups was faster than that in the model group(P<0.05).The weight of sixteenth weeks Shipei native gold method of Chinese medicine low dose and middle dose group were still lower than the normal group,the difference was statistically significant(P>0.05),and the traditional Chinese medicine high dose group compared with the normal group,the difference was not statistically significant(P>0.05),compared with the model group,there was no significant difference between the low-dose group of Chinese medicine and the middle dose group(P>0.05),high dose of Chinese medicine group were higher than those of model group,the difference was statistically significant(P<0.05).Gross observation:the lung volume of the normal control group was normal,the surface was smooth and elastic.In the model control group,the lung volume of rats in the model control group increased compared with that in the normal group,while the lung expanded and the elasticity decreased.Strengthening earth to generate metal decoction group of rat lung volume and lung was expansive situation has improved,some rats lung volume is normal,the surface is smooth,more flexible.Light microscope:the normal control of bronchus and bronchi rats structure clear,epithelium is pseudostratified ciliated columnar epithelium with goblet cells,no cilia lodging,no gland hyperplasia and no obvious inflammatory cell infiltration;alveolar ducts and alveolar structure clear,no expansion and contraction,no capillaries in the alveolar walls the dilation and congestion,alveolar wall was thickening,alveolar wall and alveolar cavity no inflammatory cell infiltration.The rats in the model control group alveolar expansion,alveolar partition narrowing or adjacent alveolar fracture,fused into a big cavity,alveolar wall pressure,the capillary bed to reduce the number of pulmonary arterioles smooth muscle hyperplasia and hypertrophy,small bronchi and bronchioles showed chronic inflammatory cell infiltration,wall smooth muscle hypertrophy significantly,disordered the main bronchial mucosal folds increase long,goblet cell hyperplasia,wall and surrounding a large number of inflammatory cell infiltration,smooth muscle hypertrophy.Compared with the model group,the alveolar structure of the low dose group was relatively complete,partial alveoli dilated,smooth muscle hyperplasia and hypertrophy were seen in the small pulmonary artery wall,bronchitis cells infiltrated at all levels,thickening of the tube wall,and goblet cell proliferation of the bronchial mucosa.Compared with the model group,the structure of pulmonary alveoli was relatively complete,and the pulmonary artery wall showed smooth muscle hyperplasia and hypertrophy,and the bronchitis cells decreased significantly,and the wall of the tubes had no obvious thickening.Compared with the model group,the alveolar structure of the high dose group of Chinese medicine was relatively intact,the bronchitis cells in all levels decreased significantly,and the wall of the tube had no obvious thickening.2.2 Effect of strengthening earth to generate metal decoction on BALF cell count in COPD ratsThe study found that the total leukocyte count was highest in COPD model group,Chinese medicine high,medium and low dose group compared with the model group significantly decreased(P<0.05),that strengthening earth to generate metal decoction has obvious inhibitory effect on inflammatory cells in COPD rats;cell counts in each group,were mainly compared with macrophages.Normal group,macrophage percentage of model group decreased significantly(P<0.01),and lymphocyte,neutrophil and eosinophil percentage were significantly increased(P<0.01),compared with the model group,Chinese medicine high dose group of mononuclear macrophage cells increased significantly(P<0.01),Chinese high,medium and low dose group the percentage of lymphocytes decreased significantly(P<0.01),Chinese medicine high dose group significantly decreased the percentage of neutrophils(P<0.01),Chinese medicine high dose group of eosinophil percentage decreased significantly(P<0.01),suggesting that cigarette smoking combined with intratracheal instillation of lipopolysaccharide induced rat model can reduce the percentage of mononuclear macrophages,lymphocytes,neutrophils and eosinophils were significantly increased,while Peitu Shengjin medicine can be significantly increased in COPD rat macrophage percentage,and decrease of lymphocytes,neutrophils and eosinophils eosinophil percentage has obvious inhibitory effect on COPD secretion of rat inflammatory cells;different dose groups influence on inflammatory cells in rats in a dose dependent manner.High dose strengthening earth to generate metal decoction group has inhibitory effect on leukocyte secretion,promote the secretion of macrophage,the lymphocyte,neutrophils and eosinophils inhibits the secretion function better than the low dose group.2.3 Effect of strengthening earth to generate metal decoction on pulmonary function in COPD ratsThe pulmonary function tests were performed at sixteenth weeks,including PIF(inspiratory peak flow rate),PEF(expiratory peak flow rate),VC(vital capacity),Cdyn(lung dynamic compliance),FRC(forced vital capacity)and MV(minute ventilation volume).The results showed that compared with normal rats,rats in the model group FRC,Cdyn,VC value decreased significantly,the difference was statistically significant(P<0.05),suggesting an increased,long-term smoking treated rats airway resistance,airway obstruction,lung compliance decreased,consistent with the clinical features of patients with COPD,can to provide reliable data for the research of COPD.The high reaction state of Chinese medicine group can obviously improve the airways of COPD mice,the difference was statistically significant(P<0.05 or P<0.01),effects of the same dose showed certain dependencies,high doses of earth to improve the strongest gold prescription,the prescription dose in the earth in the effect of gold second,improve the effect of low dose of spleen invigorating prescription is weak.2.4 Effect of strengthening earth to generate metal decoction on COPD rats serum IL-17,IL-23The study found that compared with normal group,the content of IL-17 in serum of COPD rats in the model group increased significantly(P<0.01),suggesting that the expression of IL-17 in COPD rats compared with normal rats increased,Chinese high,medium and low dose group rats serum IL-17 content compared with the model group decreased significantly(P<0.05),description of earth gold Chinese medicine has obvious inhibitory effect on inflammatory cytokines of COPD rats.The inhibitory effect of high and medium dose groups of Chinese medicine on the expression of IL-17 was better than that of low dose group(P<0.05),while the high dose group of Chinese medicine had the same effect.2.5 Effect of strengthening earth to generate metal decoction on IL-17 and IL-23 on alveolar lavage fluid in COPD ratsThe study found that compared with normal group,COPD model group rat bronchoalveolar lavage fluid IL-17 were significantly higher(P<0.01),suggesting that the expression of IL-23 in COPD rats compared with normal rats increased,high dose of Chinese medicine group rat bronchoalveolar lavage fluid IL-17 content compared to the model group decreased significantly(P<0.01),description of earth the gold of Chinese medicine on COPD rat alveolar lavage IL-17 expression has obvious inhibiting effect.The study found that compared with normal group,COPD model group rat bronchoalveolar lavage fluid IL-23 were significantly higher(P<0.05),suggesting that the expression of IL-23 in COPD rats compared with normal rats increased,Chinese high,medium and low dose group rat bronchoalveolar lavage fluid IL-23 content compared to model group decreased significantly(P<0.01),that of COPD alveolar lavage fluid in rats the expression of IL-23 has obvious inhibition effect.The effect of high dose group was better than that of middle dose group and low dose group(P<0.05),but there was no difference between low dose group and high dose group.2.6 Effect of strengthening earth to generate metal decoction on Th17,Treg of peripheral blood in COPD ratsThe study found that compared with normal group,COPD model group rats Thl7 expression in peripheral blood of rats were significantly increased compared with normal group(P<0.01),suggesting that the peripheral blood of Th17 rats increased compared with normal expression in COPD rats,Chinese high,medium and low dose group of rat blood Th17 expression compared to model group obviously decreased(P<0.01),indicating strengthening earth to generate metal Decoction on COPD in rat peripheral blood Th17 expression has obvious inhibiting effect.The effect of high dose group was better than that of low dose group(P<0.05),there was no significant difference between high and medium dose group and middle dose group and low dose group(P>0.05).The results indicated that no significant dose between peripheral Treg differences in the proportion of the rats in the model group,the rats in the normal group,strengthening earth to generate metal decoction combined treatment group,low dose of Chinese medicine in traditional Chinese medicine strengthening earth to generate metal decoction combined treatment(P>0.05).2.7 Effect of strengthening earth to generate metal decoction on Thl7 and Treg on alveolar lavage fluid in COPD ratsThe study found that compared with normal group,COPD model group rats BALF Th17 expression compared with normal group rats were significantly increased(P<0.01),suggesting that the expression of Th17 in bronchoalveolar lavage fluid in COPD rats compared with normal rats increased,Chinese medicine high dose group rat bronchoalveolar lavage fluid Th17 as contrast model group decreased significantly(P<0.05),indicating strengthening earth to generate metal expression has obvious inhibitory effect on COPD rat alveolar lavage fluid Th17.The effect of high dose group was better than that of middle dose group(P<0.05).The study found that compared with normal group,COPD model group rats BALF Treg expression compared with normal group rats decreased significantly(P<0.05),suggesting that the expression of Treg in bronchoalveolar lavage fluid in COPD rats compared with normal group rats decreased,Chinese high,medium and low dose group rat bronchoalveolar lavage fluid Th17 expression comparison model there was no significant difference(P>0.05).2.8 Effect of strengthening earth to generate metal decoction on lung tissues of COPD rats,Thl7 TregThe study found that compared with normal group,compared with normal group significantly increased the expression of Th17 in lung tissue of rats in COPD model group(P<0.05),suggesting that the expression of Th17 in lung tissues in COPD rats compared with normal rats in traditional Chinese medicine,low dose group of lung tissue expression of Th17 in rat model group were compared decreased(P<0.01),indicating strengthening earth to generate metal decoction has obvious inhibitory effect on the expression of Th17 in lung tissue of COPD rats.There was no significant difference between the dose groups(P>0.05).The study found that compared with normal group,compared with normal group rats significantly decreased the expression of Treg in lung tissue of rats in COPD model group(P<0.01),suggesting that the expression of Treg in lung tissues in COPD rats compared with normal group rats decreased,Chinese medicine high dose group in the lung tissue of rats in model group increased significantly compared with the expression of Treg(P<0.01),indicating strengthening earth to generate metal decoction has obvious effect on promoting the expression of Treg in lung tissue of COPD rats.There was no significant difference between the high dose group and the medium dose group(P>0.05).Conclusions1.Strengthening earth to generate metal decoction combined with conventional western medicine treatment compared to COPD stable lung in patients with spleen deficiency syndrome and Western medicine treatment,can effectively alleviate the symptoms of COPD patients,especially in cough,fatigue,abnormal stool.2.Strengthening earth to generate metal decoction could improve the quality of life of patients,especially in alleviating the effects of respiratory symptoms and reduce the disease of patients.3.Strengthening earth to generate metal decoction can to some extent improve the ability of daily activities.4.In the COPD model mice,the activation of Th17 cells and the high expression of upstream factor IL-23 were observed.Thl7 mediated IL-17 expression increased in bronchial mucosa and epithelial cells,stimulated other inflammatory cells,increased airway mucus secretion,and caused airway obstruction and remodeling.5.Strengthening earth to generate metal decoction could significantly improve the COPD rat model of the general condition,lung function and lung tissue damage,high dose effect is better than the low dose group.6.Strengthening earth to generate metal decoction can increase the expression of Treg cells,decrease Th17 related cytokines IL-17,IL-23 expression,and the balance in the peripheral blood of Th17/Treg ratio,reduce the inflammation of COPD rats,the high and middle dose group is better than the low dose group.7.Strengthening earth to generate metal decoction especially high dose group can reduce the expression of Th17 in peripheral blood,thereby reducing the ratio of Th17/Treg in the lung tissue,alleviating lung inflammatory reaction. |
PDF Full Text Request