Idiopathic Pulmonary Fibrosis TCM Syndrome Study And Experimental Study Of Youguiyin Intervening Pulmonary Fibrosis Rats

Idiopathic pulmonary fibrosis(EPF)is chronic and irreversible respiratory system disease with high morbidity and mortality,however,effective treatment and methods remain insufficient currently.Recently,although traditional Chinese medicine(TCM)therapy has growing advantages and characteristics in preventing and treating IPF,more theoretical research,clinical trial and experimental research need to be performed to support TCM therapy's objectivity.Yougui drink is a representative prescription,which has warmly invigorating kidney yang function,originated from 'Jing-yue Quanshu'.In previous clinical practice,this drink could relieve IPF patients' clinical signs and improve their quality of life,while,its underlying mechanisrm remains elusive.Thus,the distributing characteristics of IPF TCM syndromes are researched and effectiveness of'kidney deficiency' are confirmed in present clinical study,afterwards,in order to provide evidence for clinical treatment,the curative effect and mechanisms of yougui drink are evaluated and discussed respectively by fundamental rat experiment.ObjectivesFirslty,Study of syndrome characteristic of IPF.Investigating clinical data of 137 IPF patients to research the distributing characteristics of IPF TCM syndromes by corss-sectional stuy.Based on IPF syndromes' distributing regularity,further research the correlation between related lung and kidney deficiency syndromes and objective physical and biochemical indexes of IPF disease.Beside,ascertaining the correlation between mutual promotion among lung and kidney theory and IPF morbidity.Secondly,Research of underlyi,ng mech.anism of yougui drink for treating IPF.based on the mutual promotion among lung and kidney theory,therapeutic experiment of using yougui drink treating rat model of IPF disease will be performed.Two aspects,inflammatory reaction is modulated by PGE2 receptor and apoptosis is mediated by endoplasmic reticulum stress,are discussed to elucidating mechanism of preventive effect of yougui drink.MethodsFirstly,Chinical study.137 IPF patients who meet the inclusion criterion were recruited,and clinical data were analyzed in SPSS24.0 software.Characteristics of IPF TCM syndromes were discriminated according to clinical data.Depending on former results,kidney deficiency and lung deficiency were combined together to detecting the correlation between related lung and kidney deficiency symptoms and objective physical and biochemical indexes of IPF disease by method of Logistic stepwise regression analysis.Secondly,Experimental research.Experimental model of IPF were constructed by bleomycin aerosol inhalation.After models were constructed successfully,model control,pirfenidone therapy,low dose yougui drink therapy and high dose yougui drink therapy were grouped by random number table method,a matched normal control were set also.After 14 days model processing,every group were treated by western and herbs respectively,except the model and control groups were just given water.All rat groups were performed pulmonary function test and mercy killed for further blood gas analysis and lung tissure research after20 days treatment.Using hematoxylin and eosin stain and masson trichrome staining methods to observe lung pathomorph and histology form of every group,hydroxyproline content was detected by acid hydrolysis,what's more,Prostaglandin E2 in blood and lung tissue of every group were observed by ELISA,besides,expression and distribution of EP2,EP3 in lung tissue were also researched by immunohistochemical method.The expression of Caspase-3,CHOP and GRP78proteins in lung tissue were tested by Western Blot method,moreover,the mircro-RNA of CHOP and GRP78 were also analyzed by real-time quantitative PCR method.ResultsFirstly,Clinical study results sectionIn137 IPF patients,Qi deficiency of lung and kidney syndrome had a maximum proportion(29.93%).The rest ranking form high to low was Qi yin deficiency of lung and kidney,phlegm-heat accumulated in lung,pulmonary stagnation of phlegm,Qi deficiency of lung and yin asthenia generating intrinsic heat syndromes.Each syndrome had a statistical difference in age and duration of disease(p<0.05).Notably,patients with Qi deficiency and Qi yin deficiency of lung and kidney were older than others;In pulmonary function test,Qi yin deficiency of lung and kidney syndrome had significant lower FVC%than other syndromes except Qi deficiency of lung and kidney(p<0.05).Qi deficiency of lung and kidney syndrome had lower level in DLCO%than other syndromes except Qi yin deficiency of lung and kidney,(p<0.05);In blood gas analysis test,Qi deficiency of lung and kidney syndrome was lower than lung Qi deficiency,phlegm-heat accumulated in lung,pulmonary stagnation of phlegm syndromes in PaO2 level(p<0.05);Male,age over50 and had a experience of smoking had a significant correlation with IPF(p<0.05).moreover,FVC%and PaO2 had negative correlation with lung and kidney deficiency syndrome,however,lung and kidney deficiency syndrome maybe diagnosed with lower FVC%and PaO2.Secondly,experimental research sectionI,Morphology of lung tissue.Compared with normal control group,the model group showed alveolus structure is malalignment and collapse,and interstitial infiltration infiltrated by a large number of inflammatory cells and massive deposition of collagen fibers were found in rat lung tissue.Yougui drink could depress the lung tissue changing from consolidation zone to non-consolidation zone,reduce collagen fibers deposition.?,Content of HYP.Compared with normal control group,content of HYP in model group was significant improved(p<0.05),however,there was no significant changed contents of HYP in pirfendione group and high dose herb group.Compared with model group,the low dose herb group didn't altered,while,high dose herb and pirfendione groups were significantly reduced in content of HYP(p<0.05).?,pulmonary function and blood gas analysis.Compared with normal control group,level of FVC.?FEF25%?FEF50%?FEF75%and MMF significantly decreased(p<0.05),but ratio between FEV2 and FVC%were significantly improved(p<0.05);Compared with model group,FVC level of pirfenidone,low dose and high dose group were significantly increased(p<0.05).There was no significant difference between all herb groups and pirfenidone group in FVC;low dose group could improve FEF25%?FEF50%?FEF75%and MMF level of model group(p<0.05);No significant changed blood gas analysis factors were detected in each group,but high dose and model group showed PO2 and SO2 had a rising trend.IV,lung mass and its coefficient.Compared with normal control group,lung mass and lung coefficient were significantly improved(p<0.05)in model group,and the weight were significantly decreased(p<0.05).Compared with model group,the lung mass and its coefficient were significantly decreased(p<0.05)in low dose group,while,low dose group didn't found any significant difference.V,PGE2 in blood and lung tissue.Compared with normal control group,PGE2 in blood and lung tissue improved significantly in model group(p<0.05);Compared with model group,PGE2 in blood and lung tissue decreased significantly in low dose group(p<0.05),only PGE2 in blood decreased significantly in high dose group(p<0.05).There was no significant PGE2 difference found in lung tissue.VI,Expression of EP2 and EP3.Small amount lung cells with colored EP2,except EP3,were detected in normal control group.In lung consolidation area of model group,AECIIs found many EP2 expression.Majority of macrophage with colored EP3 were detected in alveolar duct and alveolar cavity.Compared with normal control,positive cells with EP2 and EP3 expression significantly increased in model group(p<0.05);EP2 was weakly positive expression in lung consolidation area of low dose group,but macrophage with colored EP3 significantly increased.Little enhanced EP2 were found in high dose group,while,the EP3 had similar expression with model group;Compared with control group,cells with EP2 significantly decreased(p<0.05),and EP3 were found significant increased(p<0.05),however,no changed EP2 and EP3 were found in high dose group.VII,GRP78 and CHOP expre,ssion,Compared with normal control,model group showed significantly increased GRP78 and CHOP protein(p<0.05).Compared with model group,decreased CHOP and GRP78protein were significantly detected in high dose group(p<0.05);As for mRNA expression,CHOP and GRP78 had a rising trend in model group compared to normal control,while,compared with model group,CHOP were found decreased in high dose group,and GRP78 were found decreased in low and high dose group.Any significant statistical difference were not found.VIII,C.aspase-3 protein expression.Compared with normal control,Caspase-3 protein increased in model group(p<0.05);In high dose group,Caspase-3protein decreased compared to normal control group(p<0.05).ConclusionsFirslty,lung and kidney deficie.ncy is the most common TC.M syndromes of IPF,notably,kidney deficiency clinical sig-ns are often co-occurred.Clinical study indicated lung and kidney deficiency is the most common TCM syndromes of IPF,notably,kidney deficiency clinical signs are often co-occurred.Moreover,each syndrome has concurrent blood stasis syndrome.Pathological factors of'deficiency,sputum and stasis'can be concluded as central pathogenesis for this disease,dysfunction in lung and kidney organs are taken main responsibility for deficiency factor,and lung and kidney deficiency are the fundamental causality for IPF onset.Secondly,pulmonary function deline in defrciney of lung and kidney syndrome.Qi deficiency and Qi yin deficiency of lung and kidney syndromes are distributed in aged patient.Qi yin deficiency of lung and kidney patients are accompanied with FVC decline,and Qi deficiency of lung and kidney patients are associated with DLCO%decline.Patient whose age over 50,male are susceptible to becoming lung and kidney deficiency syndrome,furthermore,decline in FVC%and PaO2 had significant correlation with this syndrome type.Thridly,Yougui driink could prevent durat.ion of disease by influencing prostaglandin E1 function and imroving apoptosis induced by endoplasmin reticulum stress.Yougui drink could relieve dysfunction of lung,and improve alveolar gas exchange function,moreover,reduce inflammatory infiltration and collagen deposition in lung tissue.One effective mechanism maybe having anti-inflammatory function by reducing content of PGE2,depressing the expression of EP2 in AEC IIs,adding expression of EP3in macrophage in lung tissue,another mechanism maybe delaying the progression of pulmonary fibrosis by modulating expression of GRP78and CHOP protein,improving lung cell apoptosis induced by ERS function.