| Objective1.To evaluate the clinical efficacy of Bushenyifeixiaozheng decoction in the treatment of pulmonary interstitial fibrosis.2.To investigate the mechanism of Bushenyifeixiaozheng decoction on interfer on JNK signaling pathway in ERS of pulmonary fibrosis rats.3.To investigate the molecular mechanism of Bushenyifeixiaozheng decoction on ERS and its apoptosis in human A549 cells.Methods1.This study has carried out the clinical observation of the treatment of pulmonary interstitial fibrosis with Bushenyifeixiaozheng decoction.According to Western diagnostic criteria and TCM syndrome diagnosis criteria screening cases,a total of 13 cases were collected,3 months for a course of treatment.the symptoms of TCM syndrome,mMRC dyspnea,cough symptom score,HRCT score,6-minute walking test and St George's breathing questionnaire score were evaluated before and after treatment to observe the clinical efficacy of the patients of fibrosis.2.SD rats were randomly divided into blank group,model group,positive drug group,traditional Chinese medicine prevention group,traditional Chinese medicine treatment group and traditional Chinese medicine control group.Except for blank group,other groups were treated with bleomycin-induced pulmonary fibrosis model.In the traditional Chinese medicine prevention group and the traditional Chinese medicine control group,the rats 12.68g/(kg · d)of Bushenyifeixiaozheng decoction and the control group were give the same amount of normal saline.After the model was established for 28 days,the rats were sacrificed and the rats in the Chinese medicine prevention group were sacrificed.The effect of prophylactic administration on pulmonary fibrosis was observed.After successful modeling,the positive drug group was given Pirenidone 53.57mg/(kg · d),traditional Chinese medicine treatment group and traditional Chinese medicine control group were treated with Bushenyangfei decoction 12.68g/(kg · d),and blank group and model group were given equal volume of normal saline for 28 days.HE staining was used to observe the pathological changes of lung tissue in each group.Real-time PCR and Western-blot were used to detect the expression of JNK signal pathway related factors GRP78,IRE1?,p-JNK,TRAF2 gene and protein content in lung tissue.3.After incubation for 24 hours,the A549 cells were divided into normal group,model group,control group,low concentration group of traditional Chinese medicine and high concentration group of traditional Chinese medicine.The normal group continued to be cultured in basal medium,the model group to basal medium+TGF-?1(5ng/ml),the control group to the basic medium+0.1%TGF-?1(5ng/ml)+0.02%TGF-?1 inhibitor(2umol/L),low concentration group of traditional Chinese medicine to the basic medium +TGF-?1(5ng/ml)+0.1%Chinese medicine lyophilized powder(100ug/ml),high concentration of traditional Chinese medicine to the basal medium+TGF-?l(5ng/ml)+0.1%Chinese medicine lyophilized powder(200ug/ml).The expression of ATF6-?,GRP78,p-JNK,PERK,TRAF2,Caspase-12,CHOP and CAMK ? in A549 cells were detected by Western-blot.Results1.After 3 mouths'treatment of bushenyifeixiaozheng decoction,in disease comprehensive effect progressive in 1 case,stable in 11 cases,ineffective in 1 case,the total effective rate was 92.3%.in TCM symptoms,clinical cure in 0 cases,markedly effective in 3 cases,limited to 8 cases of disease,The effective rate was 84.6%.compared with before treatment,the patient had no symptoms of daytime and nighttime cough symptoms(P<0.01).The effective rate was 84.6%in mMRC dyspnea grade,(P<0.01).compared with before treatment,There was no significant difference between in HRCT(P>0.05).Before treatment,the patient had a 6-minute walking distance improvement(P<0.05).compared with before treatment,patients'athletic endurance and activity Limited had been improved(P<0.05),and the impact of disease on patients had ruduced(P<0.05).2.HE staining showed that the lung tissue of the blank group was normal.The alveolar structure of the lung tissue of the model group was damaged,the fibrosis was associated with the formation of the cell lung,the fibrosis of the scar,the patchy distribution,And the fibroblasts had a certain degree of hyperplasia.The alveolar structure of the traditional Chinese medicine control group was complete and a small amount of fibroblast hyperplasia was higher than that of the model group.The degree of fibrosis was significantly higher than that of the control group Significant improvement.The result of Real-time PCR showed that the expression of GRP78,IREla,p-JNK and TRAF2 in the lung tissue of the rats in each group were significantly higher than those in the blank group.The expression of GRP78,IRE1?,p-JNK and TRAF2 in the lung tissue of the model group increased(P<0.05).Compared with the model group,there was a significant difference in the expression of IREla gene in control group(P<0.05).Compared with the model group,there was a significant difference in the expression of GRP78 and TRAF2 in the lung tissue of the traditional Chinese medicine group(P<0.05).Compared with the model group,the expression of GRP78,IRE1? and TRAF2 in the control group was lower than that in the control group(P<0.05).The result of western-blot showed that the levels of GRP78,IRE1?,p-JNK and TRAF2 in the lung tissue of each group were higher than those in the blank group(P<0.05).Compared with the model group,The expression of IRE1? was lower in traditional Chinese medicine prevention group and control group,(P<0.05).The expression of GRP78 and IRE1? in traditional Chinese medicine treatment group was lower than that in the model group(P<0.05),The contents of GRP78,IREla and p-JNK in the traditional Chinese medicine control group were significantly lower than those in the model group(P<0.05 or P<0.01).3.Compared with the nomal group,the expression of ATF6-a,GRP78,p-JNK,PERK,TRAF2,CAMK?,Caspase12 and CHOP in the model group was significantly higher than that in the model group(P<0.05).Compared with the model group,the expression of ATF6-ahad a significantly reduction(P<0.01),and The expression of GRP78,p-JNK,caspase12 and CHOP was decreased in control group(P<0.05).Compared with the model group,The expression of ATF6-a and caspasel2 was significantly decreased in the low concentration group of traditional Chinese medicine(P<0.01),and the expression GRP78,CAMK ? CHOP(P<0.05).The expression of p-JNK protein was significantly lower than that of model group in the high concentration group of traditional Chinese medicine Conclusion(P<0.01),and the expression of TRAF2,CAMK?(P<0.05).There was no significant difference in the expression of caspase12 and CHOP,but the decrease trend was not observed.Conclusions1.Bushenyifeixiaozheng decoction on pulmonary interstitial fibrosis have a certain clinical efficacy,can improve the patient's cough,dyspnea clinical symptoms,to maintain stable condition,to extend the patient 6 minutes walking distance,improve patients' athletic endurance,improve patient activity Limited,to reduce the impact of disease on patients,improve patient quality of life.2.The regulation of JNK signal pathway by reducing the level of GRP78,IRE1?,p-JNK and TRAF2,the reduction of endoplasmic reticulum stress and the reduction of apoptosis are one of the mechanisms of Bushenyifeixiao decoction to delay pulmonary fibrosis.3.By down-regulation of ATF6-?,GRP78,p-JNK,TRAF2,Caspase-12,CHOP and CAMK ? to regulate JNK,CHOP,Caspase-12 signaling pathway and Endoplasmic reticulum stress is one side of the mechanism of delaying pulmonary fibrosis. |
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