Qi Dan Hua Xian Fang On The Intervention Of Pulmonary Fibrosis In Rats And Its Mechanism Study

Objective:Pulmonary Fibrosis,with its increasing incidence and unknown cause,is becoming a severe threat to human health with no effective cure and high death rate.The clinical research has already proved that traditional Chinese herbal treatment can effectively ameliorate its symptom,considerably improve the victims' life quality and substantially reduce its deaths.Therefore,it is of necessity and significance to explore the traditional herbal treatment of pulmonary interstitial fibrosis.Qi Dan Hua Xian Fang(Herbal Prescription to dissolve pulmonary interstitial fibrosis with Qidan,astragalus & moutan bark) takes considerable clinical effects in treating pulmonary interstitial fibrosis at various stages by strengthening and supplementing the flow of Qi,promoting blood circulation and dissipating the phlegm. The project is designed to verify and assess the effects of Qi Dan Hua Xian Fang(referred to as QDHXF) via rodent experiment,thus exploring its possible operation ways and mechanism.The project is conducted by observing the effects of QDHXF on the rodents with pulmonary fibrosis induced by bleomycin.Based on QDHXF's effects on the matrix metalloproteinase system and the cytokine influencing the metabolism of extracellular matrix,the observation is focused on the pathomorphism and ultrastructure of lung tissues,the matrix metalloproteinase and its tissue inhibitors,the content of cytokine PTGFβ1 and the content of lung tissue HYP.The animal experiment is observed to give an objective assessment and evaluation of QDHXF's effects and provide a possible explanation for its operation ways and mechanism.Methods:144 healthy Wistars,weighing 200±20g,are divided into six groups of 24 members at random,i.e.The normal group,the model control,the group treated with prednisone,the group treated with high-dosed QDHXF,the group treated with medium-dosed QDHXF and the group treated with low-dosed QDHXF.Except the normal group,the other five groups are set with the model of pulmonary fibrosis by intratracheal instillation of bleomycin.From the second day on,the three groups treated with QDHXF are supplemented with the high dose of 2g/ml,medium dose of 1g/ml and small dose of 0.5g/ml.The group treated with prednisone is supplemented with prednisone acetate suspension of 0.32mg/ml.The normal group and the model group are supplemented with 10ml/kg·bw normal saline.8 mice will be executed at random on the 7^th,the 14^th and the 28^th day for observation.The research is focused on(1) the pathomorphism and ultrastructure change of lung tissues via HE dyeing and optical microscope(2) the change of lung coefficients(3) the content change of lung tissue TNF-αand cytokine CTGF via chemical analysis of immune tissues (4) the change MMP-9 and TIMP-1 of blood serum via ELISA testing(5) the change of the NO and MDA level of lung tissues and SOD activity via alkali hydrolysis of samples(6) the change of lung tissue TGFβ1 via RT-PCR testing(7) the effects of QDHXF on the above-mentioned indexes.Results:(1) QDHXF can noticeably reduce the lung coefficients of experimental pulmonary fibrosis(P＜0.01)=.The high-dosed QDHXF group has abetter effect than the low-dosed one(P＜0.05)=.There is no significant differentiation among the group treated with prednisone,the group treated with high-dosed QDHXF and the group treated with medium-dosed QDHXF(P＞0.05).The microscopic observation of the pathomorphism of lung tissues indicates that in comparison with the model group,there are various improvements among the prednisone group and the three groups treated with QDHXF.Significant differences in the infection of pulmonary alveoli are presented among different groups(P＜0.01 or 0.05),listed in descending severity as the model group,the low-dosed QDHXF group,the medium-dosed QDHXF group,the prednisone group,the large-dosed QDHXF group and the normal group.There are also substantial discrepancy in the pulmonary fibrosis(P＜0.01 or 0.05),listed in descending severity as the model group,the prednisone group,the low-dosed QDHXF group,the medium-dosed QDHXF group,the large-dosed QDHXF group and the normal group.(2) Compares with the normal control group,various time other each group of big mouse lung organizes CTGF,TNF-αexpression obvious markup(P＜0.01).Compares with the model control group, various time spot,the strong loose group,QDHXF middle monitoring team CTGF,TNF-αthe expression is lower than the model control group(P＜0.01 or 0.05),QDHXF small monitoring team slightly is lower than the model control group,but comparison difference non-statistics significance(P＞0.05).(3) Compared with the normal group,the model group has a conspicuous rise in the MDA and NO contents of lung tissues at different times,reaching a summit on the 28^th day.At all times, the prednisone group and the three groups treated with QDHXF has conspicuously lower contents of MDA and NO than those of model group at the corresponding periods(P＜0.01).Among groups,the three QDHXF groups show negligible differences on the 7^th and 14^th day; however,there is a significant difference between the high-dosed group and the low-dosed group on the 28^th day.Compared with the model group,the high-dosed QDHXF group has substantially higher SOD activity and lower MDA content in their lung tissues.(4) Compared with the normal group,the model group has a notable rise in MMP-9 and TIMP-1 contents in their blood serum on the 7^th,14^th and 28^th day(P＜0.01).Compared with the model group, the prednisone group and the three groups treated with QDHXF has an obvious decrease in MMP-9 and TIMP-1 contents(P＜0.01 or 0.05).(5) All experimental groups have TGF-β1 presentation on the 7^th 14^th and 28^th day.Compared with the normal group,the model group has a significant rise on the 7^th day(P＜0.01) and reaches its summit on the 14^th day.With its relative fall on the 28^th day,it is still much higher than the normal group at the corresponding period(P＜0.01).Compared with the model group,the prednisone group and the three QDHXF groups have conspicuous fall(P＜0.01).Among the three QDHXF groups,the TGF-β1 of the high-dosed group is considerably lower than that of the low-dosed group(P＜0.05).Conclusion:QDHXF has significant effects on the prevention and treatment of the pulmonary fibrosis induced by bleomycin,which is likely to be realized by reducing MMP-9 and TIMP-1 in blood serum,regulating MMP-9/TIMP-1 system to its balance,reducing CTGF and TNF-contents, cutting MDA and NO contents in lung tissues, strengthening SOD activity and reducing TGF-β1 in lung tissues. To sum up,modify the over sediment of extracellularmatrix,thus effectively protecting the normal lung tissues,preventing and treating the.