The Zhongjing Treatment Feiwei Dialectical Law And Related Experimental Study

Idiopathic pulmonary fibrosis(IPF) is an chronic inflammation Interstitial lung disease ,it's marked pathologic change is interstitial pneumonitis.The main clinical manifestation of IPF is diffuse airsacculitis, alveoli of lung disorder,pulmonary fibrosis.The pathogenetic condition of IPF patient deteriorate,many IPF patients result in death because of respiratory failure.Now ,the drugs to treat IPF include glucocorticosteroid, immunodepressant /cytotoxic drug et al.These drugs have grave adverse reaction If patients long-term take these medicines,will cause body immunity to descend and secondary infection.So,it is investigation hot spot to search eutherapeutic, little side effect drugs.IPF don't have corresponding syndrome name in traditional Chinese medition (TCM),it is relegated to"pulmonary arthralgia""pulmonary paralysis""the syndrome of dyspnea"by many traditional Chinese physicians.I think, IPF is belong to"pulmonary paralysis"of TCM.Traditional Chinese physicians of past dynasties have a great deal therapeutic methods to pulmonary paralysis,but all scatter in writings of traditional Chinese physicians of past, there is no system summary and induction. The theory research of my topic will systematic summarize therapeutic methods of traditional Chinese physicians to pulmonary paralysis, and bind to modern medicine, to explain some promiscuous questions , supply theory foundation for treat pulmonary paralysis by traditional Chinese medition.The empirical study of my topic will tracing latest development of international, to cooperate with Yale university, apply Jia Wei Jie Geng Decoction to cure IPF model mice, observe Jia Wei Jie Geng Decoction's curative effect to IPF model mice , explore action mechanism of Jia Wei Jie Geng Decoction cure IPF , survey content change of bone morphogenetic protein-4 (BMP-4) and bone morphogenetic protein-7 (BMP-7), supply experiment foundation for treat pulmonary paralysis by classical prescription..1.Theory study1.1 Systematic summarize therapeutic methods of Traditional Chinese physicians to pulmonary paralysis.To look up TCM documents such as internal medicine, prescription, medical record, medical talks et.al, analysis and summary these TCM documents, probe experience and method of antiquity great physician treat pulmonary paralysis, reveal rule and achievement of TCM in diagnosis and treat pulmonary paralysis.1.2 Systematic summarize Biangzheng pattern of Zhongjng treat pulmonary paralysis.2. Empirical study2.1 survival rate experimentTake thirty C57BL/6 mice, random divided to the Chinese medicine survive group and the Bleomycin A5 survive group,all mice instillation Bleomycin A5 in airtube to make IPF model , the mice of the traditional Chinese medicine survive group intragastric administrate Jia Wei Jie Jeng Decoction,the mice of the Bleomycin A5 survive group intragastric administrate isotonic Na chloride, observe the survival condition of two group mice, depict survive curve, put to death all mice in twenty nine day after make IPF model .Experiment result indicate the survival rate of Chinese medicine survive group is 46%,survival rate of the Bleomycin A5 survive group is13%,this result indicate that the Jia Wei Jie Jeng Decoction has noticeable therapeutical effect to IPF model mice. 2.2 The pathological observation experiment of IPF mice model pulmonary tissue sampleTake eight mice part right lung in every group after make IPF model 0 day,1 day,3 day,5 day,9 day,14 day,28 day, fixed mice pulmonary tissue by 10%formalin, paraffin imbedding and slice, HE dyeing, to observe the pulmonary tissue pathological change of IPF mice model. The experiment result indicate that the Jia Wei Jie Geng Decoction group has marked therapeutic action to IPF mice model. Mice pneumonitis symptom of the Jia Wei Jie Geng Decoction group lessen than the Bleomycin A5 group, inflammatory cell grow downwards,the Jia Wei Jie Geng Decoction group mice have not appear pulmonary fibrosis.2.3 Bronchial lavage experimentTake two hundred C57BL/6 mice, random divided to three group: the Jia Wei Jie Geng Decoction group ,the Bleomycin A5 group and the isotonic Na chloride group.The isotonic Na chloride group has 60 mice, the Jia Wei Jie Geng Decoction group has 70 mice, the Bleomycin A5 group has 70 mice, Mice of the Isotonic Na chloride group don't make model, the Jia Wei Jie Geng Decoction and the Bleomycin A5 group make IPF model by instillation Bleomycin A5 in mice airtube.Take eight mice in every group after make IPF model 0 day,1 day,3 day,5 day,9 day,14 day,28 day, macroscopic observation pathological change of mice pulmonary and make bronchial lavage, calculate cell amount by decuple micro,then make Jamsa dyeing, calculate cell percentage of neutrophil, macrophage,lymphocyte.Neutrophil is main inflammatory corpusclecute of lung injury morning stage, the neutrophil percentage of the Bleomycin A5 group and the Jia Wei Jie Geng Decoction group raise up after make the IPF mice model five days, have highly significant difference with the isotonic Na chloride group, this experiment result demonstrate that mice of the Bleomycin A5 group and the Jia Wei Jie Geng Decoction group have injury of lungs and alveolitis in the morning stage after make the IPF mice model. the neutrophil percentage of the Bleomycin A5 group and the Jia Wei Jie Geng Decoction group to descend after make the IPF mice model nine days,but the descend extent of the Jia Wei Jie Geng Decoction group super than the Bleomycin A5 group, after make the IPF mice model twenty eight days, Bleomycin A5 group have significant difference with the isotonic Na chloride group,but the Jia Wei Jie Geng Decoction group have no difference with the isotonic Na chloride group. This experiment result demonstrate that the Chinese medicine promote mouse pneumonitis to assimilate and tissue repair.In the morning stage of IPF, lung alveolus macrophage could synthesis and release active oxygen mediator and tumor necrosis factorαto damage pulmonary tissue, lung alveolus macrophage still secrete various kinds cell factor such as platelet-derived growth factor, transforming growth factorβ, these cell factors control fibroblast to gather, activation, proliferate. After make the IPF mice model, Macrophage percentage of the Bleomycin A5 group and the Jia Wei Jie Geng Decoction group lower than the isotonic Na chloride group,but have no determinate tendency. Macrophage absolute number of the Bleomycin A5 group still super than the isotonic Na chloride group, Macrophage absolute number of the the Jia Wei Jie Geng Decoction group lower than the Bleomycin A5 group, demonstrate the Chinese medicine restraint the macrophagocyte multiplication in certain extent.After make IPF mice model , Lympholeukocyte percentage of the Jia Wei Jie Geng Decoction group and the Bleomycin A5 group all raise up, lympholeukocyte percentage of the Bleomycin A5 group gradually ascend, reach peak after make IPF mice model nine day, gradually descend after make IPF mice model fourteenth day,but lympholeukocyte percentage of the Bleomycin A5 group still have significant difference with the isotonic Na chloride group after make IPF mice model twenty eight day. Lympholeukocyte percentage of the Jia Wei Jie Geng Decoction group also raise up, but gradually descend after make IPF mice model five day,lympholeukocyte percentage of the Jia Wei Jie Geng Decoction group have no significant difference with the isotonic Na chloride group after make IPF mice model twenty eight day.Lymphocyte is inflammatory cell which appear after macrophage and neutrophil,but lymphocyte maintain high level in long time.our research discovery that macrophage and neutrophil grow in number precede lymphocytic infiltration. .our research also discovery lymphocyte obviously grow in number after make IPF mice model nine day,at the same time fibroblast also increase, after make IPF mice model fourteen day, fibroblast obviously increase and collogen deposition, at the same time lymphocyte is main inflammatory cell in the lung,this experiment result indicated lymphocyte relevant with fibroblast generation and collogen deposition.2.4 Survey mice left lung BMP protein level by enzyme-linked immunosorbentassayTake eight mice left lung in every group after make IPF model 0 day,1 day,3 day,5 day,9 day,14 day,28 day, Survey mice left lung BMP-4 and BMP-7 protein level by enzyme-linked immunosorbentassay, the experiment result indicate,the BMP-7 protein level of the Bleomycin A5 group mice and the Jia Wei Jie Geng Decoction group mice not so many as the isotonic Na chloride group mice,but this downtrend had been hold back by Chinese medition,so the BMP-7 protein level of the Jia Wei Jie Geng Decoction group mice begin to advance in ninth day, to achieve normal after make IPF mice model twenty eight day. The BMP-7 protein level of the Bleomycin A5 group mice constantly descend, have significant difference to compare with the isotonic Na chloride group mice. the BMP-4 protein level of the the Jia Wei Jie Geng Decoction group mice not so many as the isotonic Na chloride group mice,this downtrend had not been hold back by Chinese medition, the BMP-4 protein level of the the Jia Wei Jie Geng Decoction group have significant difference to compare with the isotonic Na chloride group after make IPF mice model twenty eight day. 2.5 Survey mice right lung BMP mRNA level by reverse transcription-polymerase chain raction.Take eight mice right lung in every group after make IPF model 0 day,1 day,3 day,5 day,9 day,14 day,28 day, Survey mice right lung BMP-4 and BMP-7 mRNA level by reverse transcription-polymerase chain raction (Rt-PCR). the experiment result indicate,the BMP-4 mRNA level of the Bleomycin A5 group mice and the Jia Wei Jie Geng Decoction group mice continuity to descend after make IPF mice model, have significant difference to compare with the isotonic Na chloride group mice.This result indicate the Chinese medition has not regulation effect to BMP-4 mRNA level of IPF model mice. The BMP-7 mRNA level of the Bleomycin A5 group mice continuity descend after make IPF mice model, have significant difference to compare with the isotonic Na chloride group mice. The BMP-7 mRNA level of the Jia Wei Jie Geng Decoction group mice descend after make IPF mice model, then slowly to ascend after make IPF mice model five day, this result indicate the Chinese medition can increase BMP-7 mRNA level of IPF model mice.The BMP-7 may the effect target of the Chinese medition cure the IPF mice model.The empirical study of our topic investigate the therapeutic action of Jia Wei Jie Geng Decoction to IPF mice model from these aspect: living state of IPF mice model; observe pathological change of IPF mice model; kinesis monitor BMP-4,BMP-7 level of IPF mice model.We can make this conclusion by these empirical study:1. The 28 day survival rate of Chinese medicine survive group super than the Bleomycin A5 survive group,this result indicate that the Jia Wei Jie Jeng Decoction has noticeable therapeutical effect to IPF model mice.2. Mice pneumonitis symptom and the injury of pulmonary of the Jia Wei Jie Geng Decoction group lessen than the Bleomycin A5 group, inflammatory cell grow downwards,the Jia Wei Jie Geng Decoction group mice have not appear pulmonary fibrosis after make IPF mice model. 3.The protein level and the mRNA level of BMP-4 all descend,the Jia Wei Jie Geng Decoction group have no effect to improve BMP-4 level of IPF mice model,but the Jia Wei Jie Geng Decoction group increase the BMP-7 level of IPF mice model, this result indicate that the BMP-7 may be the effect target of the Chinese medition to cure IPF mice model,the concrete effect mechanism need further explore.