Study On The Abnormal Expression Of MiRNAs In CD4+T Lymphocytes Of Psoriasis Vulgaris Patients Correlated With Th17Cytokine And The Intervention Effects Of ZhuHuang Granule

Objective:Using miRNA gene chip technology test miRNAs expression profiling of Psoriasis Vulgaris’CD4+T lymphocytes,Filter the different miRNAs from normal in CD4+T lymphocytes, and validate partial differential miRNAs with quantitative real-time PCR (RT-qPCR). RT-QPCR miR-155、RORytmRNA、IL-17mRNA in CD4+T lymphocytes and skin.ELISA Th17cytokine(IL-6、IL-17、IL-23) in plsam. Observed the traditional Chinese medicine ZhuHuang granules’clinical efficacy of the treatment of PV, and observed the ZhuHuang granules interventions of expression of miR-155and Thl7cytokine.Investigate the mechanism action of miR-155and Th17cytokine in PV.Explore the new role target of the traditional Chinese medicine treatment of psoriasis. Provid new theoretical and experimental basis for the Chinese medicine preventing and treating psoriasis. Methods:1.Collected4patients with PV and4healthy volunteers’ CD4+T lymphocytes and extracted RNA by Trizol method.Test miRNAs expression pedigree in CD4+T lymphocytes of PV patients and the normal by Affymetrix miRNA2.0chip.Analyze the different expression of miRNAs by clustering analysis software (cluster3.0).2. Collected20patients with PV and10healthy volunteers CD4+T lymphocytes and verify the partial differential miRNAs by RT-qPCR method.3.Collected plasma and CD4+T lymphocytes of24patients with PV and14healthy volunteers.Collected skin lesions and tissue around lesion of severe PV and normal skin of the healthy.RT-QPCR miR-155、RORytmRNA、IL-17mRNA in CD4+T lymphocytes and skin.ELISA Th17cytokine(IL-6、IL-17、IL-23) in plsam.Analyze correlation between miR-155and skin lesions severity and Th17cytokine.4.24patients with PV were treated for8weeks with ZhuHuang granules, CD4+T lymphocytes and plasma collected before and after treatment.RT-QPCR miR-155、 RORytmRNA、IL-17mRNA in CD4+T lymphocytes and ELISA Th17cytokine(IL-6、 IL-17、IL-23) in plsam.And analyze the intervention effect of ZhuHuang granules with miR-155and Th17cytokine.Results:1.There were18miRNAs differentially expressed obviously in PV patients’CD4+T lymphocytes, which13miRNAs were obviously up-regulated and5miRNAs were of them obviously down-regulated.2.Compared with normal, miR-155、miR-146a、miR-21、miR-99a、and miR-125b in PV patients’CD4+T lymphocytes were obviously up-regulated, the differences were statistically significant (p<0.05), miRNA-99a and miR-125b were obviously down-regulated, the difference have significant (p<0.05).The qRT-PCR results were in accordance with those obtained using microarray analysis.3.Compared with normal, miR-155-. RORyt and IL-17mRNA in PV patients CD4+T lymphocytes were obviously up-regulated.IL-17、IL-6、IL-23in PV patients plasma were obviously up-regulated(p<0.05).MiR-155、RORγt and IL-17mRNA in PV patients skin lesions and tissue around lesion were differentially expressed obviously compared with normal skin (p<0.05).4. The expression levels of miRNA-155、IL-17and PASI was positively correlated (p<0.05), The expression levels of miRNA-155and RORγt、IL-17mRNA、IL-17in plasm was positively correlated (p<0.05).5.After8weeks of treatment with ZhuHuang granules, PV patients’ PASI decreased significantly, and the difference was statistically significant compared with that before treatment (p<0.05), the total effective rate was79.17%.MiR-155、RORγt and IL-17mRNA in CD4+T lymphocytes after treatment were decreased compared with pre-treatment, miRNA-155and IL-17mRNA were down-regulated, the difference was significant (p<0.05).Thl7cytokines expression after treatment were compared with the pre-treatment,The result is that IL-17were down-regulated and the difference was significant (p<0.05).Conclusion: 1.13miRNAs in PV patients CD4+T lymphocytes were obviously up-regulated and5miRNAs were obviously down-regulated. The18different miRNAs may play an important role in the pathogenesis of psoriasis.2. MIR-155、RORyt and IL-17mRNA in PV patients’CD4+T lymphocytes and skin lesions were obviously up-regulated.IL-17、IL-6、IL-23in plasma were obviously up-regulated.MIR-155and Th17cytokine is important in the pathogenesis of psoriasis.3. The expression levels of miRNA-155and IL-17were 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 of PV, it may play a role through regulation of the expression levels of miRNA-155and IL-17.