Analysis The Clinical Features Of CTD-ILD And The Level Of Circulating Fibrocyte

Aim:Diffuse connective tissue disease or connective tissue disease, is one of the categoriesof disease of the rheumatic diseases, belongs to the organ specific autoimmune disease. Itincludes more than10diseases, namely rheumatoid arthritis, primary Sjogren syndrome,systemic sclerosis, polymyositis/dermatomyositis, systemic lupus erythematosus, mixedconnective tissue disease,etc. Recent studies showed that the cause of death in patientswith CTD trends to its complication. During the long-term chronic course of the disease,patients often develop a variety of complication, including heart, lung, kidney, skin, eyeand nervous system,etc. ILDs may account for15%of all CTDs patients. Symptoms arenot specific and subtle, also completely absent. Due to the limitations we have, manypatients often can be confirm the diagnosis to the disease of the disease of irreversiblephase. According to the clinical observation, it showed there is an upward trend inCTD-ILD incidence. So early diagnosis and effective treatment for the disease is thecurrent clinical needs to solve the problem.The most common in the CTD，such as RA, isa kind of chronic inflammation in synovial joints as the main performance of systemicautoimmune disease.. The cause of death in patients with RA analysis found that lunginvolvement was as high as10%~20%. Paying attention to ILD is the most common,seriously affecting the quality of life and prognosis. Circulating fibrocyte is a kind of precursor cells derived from bone marrow, a newlydiscovered leukocyte subtypes. The fibroblast from RA patients plays an important role injoint and outside the joint. Considering the source of the fibroblast, outcome, andpathological physiological activity, Circulating fibrocyte becomes one of the keypromblems. Once tissue damage, Circulating fibrocyte differentiates fibroblast ormyofibroblast in physiological and pathological process of tissue damage repair. Moreoverthe increased circulating fibrocytes secrete cytokines, chemokines and growth factors,mediate its differentiation, chemotaxis and gathered in chronic inflammatory disease.After the analysis of CTD-ILD clinical cases, we study the level of circulating fibrocytesin RA-ILD patients and CIA animal model combined BLM model, and discuss the its rolein the pathological process.Methods:1． Analysis the clinical characteristics of CTD-ILDAnalysis of inpatients from january2011to december2013.619cases of CTD-ILDinclude rheumatoid arthritis, primary Sj gren syndrome, systemic sclerosis, polymyositis/dermatomyositis, systemic lupus erythematosus and mixed connective tissue disease.Clinical data were collected, namely manifestations, serological examination, pulmonaryfunction, blood gas analysis and chest HRCT. According to clinical characters and HRCT,we classified CTD-ILD into preclinical,acute pneumonitis and chronic interstitialdisease.All data were summarized its clinical characteristics.2． Observed the level of circulating fibrocryte in RA-ILD patientsWe enrolled RA patients and RA-ILD patients from February2013to November2013Moreover, collect age, gender, disease duration, lung function, blood gas analysisand HRCT data. Flow cytometry was used to observe the level of circulating fibrocytes inRA-ILDpatients. Then combined clinical data do correlation analysis.3． Established BLM and CIA dual animal modelBased on the BLM model,we combined CIA animal model. Mice were randomlydivived into3groups. On the2,7,14,21and28day after bleomycin challenge, mice ofeach group were sacrificed. Hematoxylin Eosin (HE) staining and Sirius Red staining were used to detect the degrees of acute inflammation and fibrosis respectively. With thelung tissue hydroxyproline determination, we envaluate the onset and progress of dualanimal model.4． Observed the role of circulating fibrocryte in dual animal modelFlow cytometric analysis of circulating fibrocryte at different periods. Through thecomparison between groups, we explore whether circulating fibrocyte is involved in theoccurrence of disease and development? The lung tissue of alpha-smooth muscle actinimmunohistochemistry staining was to study whether circulating fibrocyte participate theproliferation of fibroblasts. Moreover, inflammation and fibrosis scores was to makecorrelation studies.Results:1． Clinical data of CTD-ILD has significant heterogeneityAmong619ILD patients, there was126men and493women, and male-to-femaleratio was1:3.2. Various types of patients in the clinical manifestations, serology (RF, IgGand IgM), lung function (TLC%, DLco%and FVC%) and other indicators weresignificantly different (P <0.05). With HRCT, pulmonary function tests, and serologydiagnostic strategy, we not only found preclinical patients early, but also determine theseverity of disease, which can provide valuable information for staging treatment.2． The level of circulating fibrocryte in RA-ILD patients was increased..The level of peripheral blood circulating fibrocytes in selected48RA patients werehigher than in the healthy control group, especially it increased significantly in patientswith acute exacerbation, and circulating fibrocytes of acute exacerbation patientsassociated with CRP, DAS28and DLco%Indicators. Studies suggest that in acuteexacerbation RA-ILD patients, circulating fibrocytes likely to participate in thedevelopment and progression of disease.3． Successfully established BLM and CIA dual animal model.Both two model groups showed the dynamic process from pulmonary alveolitis tofibrosis gradually. Inflammatory lesion in the lung of the single group were mostobviously on the7th and14th day. Then there was a slow rehabilitation process on the21th and28th day. The double group began presented alveoli destruction and collagen depositionon the14th day(the highest on the28th day). Pulmonary hydroxyproline contents wasobviously increased in double group compared with single group after14days.4． The CIA-ILD animal model circulating exhibits significant fibrocyte increasedThe Flow cytometry analysis revealed that an increase in the total number ofcirculating fibrocytes was observed between BLM single group and CIA-ILD doublegroup. It tended to increase at first, then subsequently decline. Level of circulatingfibrocytes were higher in CIA-ILD double group compared with BLM single group on day14,21and28(P<0.05). The immunohistochemical results showed there were moremyofibroblasts infiltrating in double group, compared with BLM single group. Circulatingfibrocytes in blood were positive correlation with pulmonary inflammation, fibrosis andHYP contents(r=0.847，0.826，0.735, P<0.01).Conclusions:After understanding the new ILD guidelines, we analysed clinical cases characteristicsin recent3years. Firstly, with HRCT, pulmonary function tests, and serology diagnosticstrategy combining,we not only found preclinical patients early, but also determine theseverity of disease, which can provide valuable information for staging treatment.Secondly,the level of circulating fibrocytes in acute exacerbation of RA-ILD patients wassignificantly increased. Lastly, aninimal model data showed CIA-ILD model presentgrowing process of chronic inflammation. Level of circulating fibrocytes were higher indual group compared with single group in advanced stages. Circulating fibrocyte in bloodwere positive correlation with degree of pulmonary inflammation, fibrosis. DuringCIA-ILD model process, circulating fibrocytes may be involved in the inflammation andfibrosis progression of lung and can provide important targets for possible interventiontreatment. Through research, we initial study the level of circulating fibrocytes, intendingto discuss its role in occurrence and progression of RA-ILD disease.