| Objective(1) We researched on some clinical risk factors which were correlated with pulmonary arterial hypertensions (PAHs) in systemic sclerosis (SSc), in order to make an early diagnosis and evaluation of SSc and to predict the prognosis in these patients;(2) We screened and verified the peripheral blood microRNA markers in SSc patients with interstitial lung diseases (ILDs);(3) We screened the serum protein spectrum of SSc patients with PAHs or ILDs.Method(1) We collected the clinical records and lab results of SSc patients in clinics and hospitalized patients in Pekin Union Medical College Hospital (PUMCH). Then we used right heart catheterization results to decide the PAH diagnosis, deviding the patients into two groups of SSc-PAH group and SSc-nonPAH group. According to the data, we used binary logistic regression and relative analysis to analyze them in order to find out the risk factors of SSc-PAH.(2) We collected the clinical records and blood samples of SSc patients in clinics and hospitalized patients in PUMCH. We screened the microRNA spectrum in these patients’peripheral blood and verified these results using real time PCR in a larger sample number. According to the statistic analysis, we found out the microRNA markers related to SSc-ILD in both plasma samples and peripheral blood mononuclear cells (PBMCs). Then, we did bioinformatics analysis to predict the target genes.(3) We screened the serum protein spectrum in a group of SSc patients with or without PAH and ILD, so as to find out the related biomarkers.Result(1) In the analysis of clinical features, we found out some items related with PAH in SSc, which were the presence of gastroesophageal refluxes (60%vs.36%, P<0.05), digital ulcers (52%vs.31%, P<0.05) and telangiectasias (64%vs.38%, P<0.05), the positivity of anti-RNP antibodies, anti-SSA antibodies and anti-SSB antibodies (respectively60%vs.18%,36%vs.18%,16%vs.4%, P<0.05), the negativity of anti-Scl-70antibodies (8%vs.50%, P<0.05), the decrease of FVC%predicted, FEV1% predicted, TLCO%predicted and TLCO/VA%predicted (respectively65%vs.34%,65%vs.37%,100%vs.74%,100%vs.60%,P<0.05), and the increase of the calculated ratio of FVC%predicted and TLCO%predicted (1.93±0.67vs.1.29±0.29, P<0.05). After the logistic regression, we found five risk factors of PAH in SSc, which were the presence of gastroesophageal refluxes and telangiectasias, the elevation of IgA levels, the positivity of anti-RNP antibodies, and the increase in the calculated ratio of FVC%predicted and TLCO%predicted.(2) In the microRNA microarray analysis, we got25differential expressed microRNA, including7up-regulated and18down-regulated ones in five SSc patients versus three healthy people. In these25microRNAs, we picked out five ones with possibilities of participations in SSc pathogenesis, which were has-miR-29b, has-miR-320a, has-miR-320b, has-miR-320c and has-miR-423-5p.Then the real time PCR results of these five microRNAs in30SSc patients versus16healthy people were as follows:in plasma, miR-320a, miR-320b and miR-423-5p were significantly down-regulated (RQs are respectively0.58±0.36vs.1.12±0.66,0.67±0.27vs.1.44±1.08,0.74±0.47vs.1.38±1.00, P<0.05), while the rest two had tendencies of down-regulation; in peripheral blood mononuclear cells (PBMCs), there were significant down-regulations of miR-29b, miR-320a, miR-320b and miR-423-5p in SSc patients with ILD (0.43±0.14vs.1.03±0.65,0.48±0.22vs.0.92±0.48,0.31±0.18vs.0.83±0.59,0.32±0.14vs.0.95±0.66, P<0.05), and a tendency of down-regulation of miR-320c; In correlation analysis, some microRNAs (miR-320a in plasma; miR-29b, miR-320a, miR-320b, miR-320c and miR-423-5p in PBMC) levels in both plasma and PBMCs showed correlations with the presence of digital ulcers (relative coefficients respectively0.603,0.436,0.547,0.470,0.728and0.513,P<0.05), and miR-320b level (relative coefficient=0.527,P<0.05) in plasma showed a correlation with the presence of telangiectasias.(3) In the serum protein microarray analysis in15SSc patients versus5healthy people, we got12differential expressed cytokins, with six ones up-regulated (GDNF, IL-26, TRA-1-81, Calcitonin, SRMS, Aldolase A) and six ones down-regulated (Fibronectin, Serpin A5, Chordin-Like2, Serpin Al, Serpin A4, C3a).Compared the SSc patients with PAH and those without, we found out five cytokins up-regulated, which were IL-17D, RYK, IL-13R alpha l, CD97and Fyn. Compared the SSc patients with ILD and those without, we found out three cytokins up-regulated, which were NET1, Netrin G2and Galanin.Conclusion(1) We found out some clinical features and laboratory results useful in the prediction of PAH in SSc, which would be the basis for making decisions of invasive exams;(2) We found out some differential expressed microRNAs in peripheral blood, which may be correlated with distinct organ involvements in SSc patients;(3) We screened out some serum cytokins which may be correlated with pulmonary diseases in SSc patients, including PAHs and ILDs. |
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