Study On The Expression Of MiRNAs In Peripheral Blood Mononuclear Cells Of Psoriasis Vulgaris Patients And The Intervention Of ZhuHuang Granule For Differentes MiRNAs And Cytokines

Objective:Using miRNA gene chip technology tested Psoriasis Vulgaris(PV) Peripheral blood mononuclear cells(PBMCs) miRNAs expression profiling, the different miRNAs expression from both abnormal and normal PBMC are screened, and validated partial differential miRNAs with quantitative real-time PCR (RT-qPCR). Using quantitative protein chip technology tested PV plasma40kinds of cytokines, the different cytokines expression from both abnormal and normal plasma are screened. At the same time, Observed the traditional Chinese medicine ZhuHuang granules clinical efficacy of the treatment of PV, and observed the ZhuHuang granules interventions of differences miRNAs and CK expression. And to investigate the mechanism action of miRNAs and CK in PV, explore the new role target of the traditional Chinese medicine treatment of psoriasis. Provided new theoretical and experimental basis for the Chinese medicine preventing and treating psoriasis.Methods:1.Collected20patients with PV and15healthy volunteers PBMCs, RNA was extracted using Trizol method, used Affymetrix miRNA2.0chip test PV patients and normal PBMCs miRNAs expression pedigree.Used clustering analysis software (cluster3.0) to analyze the differences of expression miRNAs.2. Collected20patients with PV and10healthy volunteers PBMCs, used RT-qPCR method to verify the partial differential miRNAs.3.Collected25patients with PV and15healthy volunteers plasma,used Raybiotech quantitative double-antibody sandwich protein chip technology to detect the expression of40kinds CK. Used clustering analysis software (cluster3.0) to analyze the differences of expression CK.4.25patients with PV were treated for8weeks with ZhuHuang granules, PBMCs and plasma collected before and after treatment, used RT-QPCR to test some differences andz Raybiotech quantitative double-antibody sandwich protein chip (QAH-INF-3-2) test differences CK. TO explore the correlation between the differences miRNAs and CK and severity.And analyze the intervention effect of ZhuHuang granules with differences miRNAs and CK.Results:1.There were39miRNAs differentially expressed obviously in PV patients PBMCs, which26miRNAs were obviously up-regulated and13miRNAs were of them obviously down-regulated.2.Compared with normal, miRNA-146a, miRNA-31and miRNA-192-5p in PV patients PBMCs were obviously up-regulated, the differences were statistically significant (p<0.05), miRNA-99a and miRNA-200c in PV patients PBMCs were obviously down-regulated expression were, the differencehave significant (p<0.05), consistenting with the the miRNA gene chip results.3.Compared with normal,26cytokines in PV patients plasma were differentially expressed, which18cytokines were obviously up-regulated (q<0.05) and8cytokines were obviously down-regulated (q<0.05).4. After8weeks of treatment with ZhuHuang granules, PV patients’ PASI decreased significantly, and the difference was statistically significant compared with before treatment (p<0.05), the total effective rate was80%.5differences miRNAs expression after treatment were compared with the pre-treatment, miRNA-146a were down-regulated and miRNA-99a were up-regulated, the difference was significant (p<0.05).26differences cytokines expression after treatment were compared with the pre-treatment, PDGF-BB and RANTES were up-regulated and TNF-a, MIP-1δ, TNF sRII, and ICAM-1were down-regulated, the difference was significant (q<0.05).5.The expression levels of miRNA-146a and PASI was positively correlated (p<0.05), The expression levels of miRNA-99a and PASI was negatively correlated (p<0.05). The expression levels of PDGF-BB and PASI was negatively correlated (p<0.05), The expression levels of TNF-a, MIP-1δ and ICAM-1and PASI was positively correlated (p<0.05).Conclusion:1.26miRNAs in PV patients PBMCs were obviously up-regulated and13miRNAs were obviously down-regulated. The39differences miRNAs may play an important role in the pathogenesis of psoriasis.2.18cytokine in PV patients plasma were obviously up-regulated and8cytokine were obviously down-regulated. Cytokine network is important in the pathogenesis of psoriasis.3. The expression levels of miRNA-146a, miRNA-99a, PDGF-BB, TNF-a, MIP-1δ and ICAM-1were closely related to PV skin lesions severity,which can be used as biological indicator to judge the patient’s condition and clinical efficacy.4. The Chinese medicine ZhuHuang granules have significant effection for therapy PV, it may be play a role mechanism through bidirectional regulation of the expression levels of miRNA-146a, miRNA-99a and TNFa, MIP-1δ,TNF sRII, ICAM-1, PDGF-BB, RANTES.