Study On The Differential Exppession Of MiRNA In Psoriatic Skin And The Intervention Effects Of ZhuHuang Granule

Background:Psoriasis is one of the most common clinical chronic inflammation skin diseases. Many factors influence its incidence process, involving polygenetic change, complicated progress of pathological changes. With the development of the society and the environment changes, the rate of the disease increased year by year, due to its unclear etiology and mechanism, its treatment is difficult and easy to repeat. Therefore, research for psoriasis mechanism has caused the wild attention in the field of dermatology. In recent years, with the exploring to psoriasis progressing from domestic and foreign academic circles, it has been found that many minimal RNA (miRNA) play an important role in its pathogenesis and some of them were found abnormal expression in some skin lesions, such as skin cancer, psoriasis, atopic dermatitis. At the same time, from the perspectives of gene, studying on the psoriasis diagnosis, treatment, and prevention early has become one of the main research fields in the future all over the world.Purpose:Using gene chip technology testing psoriasis lesions miRNA expression patterns, the different miRNA expression from both abnormal and normal skin tissue are screened, and the differences in miRNA target genes prediction and target genes function enrichment are analysed, so as to construct a gene function regulation web for psoriasis differentially expressed miRNA and target genes and, providing the basis for the research of diagnosis and treatment of the psoriasis at the gene level, giving the clues for the next step in miRNA target genes function and molecular markers study. At the same time, the studies on influence of the traditional Chinese medicine ZhuHuang granules on the differences in psoriasis lesions and serum miRNA patients have been carried out before and after the treatment, so that the mechanism of Chinese medicine on the psoriasis treatment was explored, and more objective basis were provided for the Chinese medicine preventing and treating psoriasisMethods:20patients with psoriasis and15healthy volunteers were recruited, the skin and tissue samples from Trizo law to extract RNA were collected. Using Affymetrix miRNA2.0chip test the miRNA expression profile of normal skin organization and patients with psoriasis lesions in the typical organization. With clustering analysis software (cluster3.0) to analyze the differences of expression miRNA, and by use real time fluorescence quantitative polymerase chain reaction (PCR) to verify part miRNA differences. Through the miRNA target genes database (miRGen3.0) forecast target genes, Gene Ontology based on database of target genes function enrichment was analysed.20psoriasis volunteers lesions samples were collected again2months after the treatment of ZhuHuang granules for miRNA chip testing. Analyse the differences and significance on the miRNA expression before and after treatment. At the same time, using the method of ELISA to test differences on the serum cells factor before and after treatment.Results:There are55miRNAs obviously up-regulated in psoriasis lesions and12miRNAs of them obviously down-regulated. The result of target genes forecast for5specific miRNA (miR-31, miR-155, miR-192, miR-139-5p, miR-497) draw about33key genes,such as AIRE, ADRA1D, AREG, POU6F1, BUB1B IKBKE, SMC3, SOCS1, DGKH, MAP3K12, OTC, REV3L, SLC26A6etc. From the function analysis, its control the original mitogen activated protein kinase signal pathway (MAPK signaling pathway), Wnt signal pathway (Wnt signaling pathway)30other major signaling pathways, mainly by adjusting the inflammation occurs, the cell apoptosis, endothelial cell proliferation and angiogenesis, new humoral immune, protein kinase C (protein kinase C, PKC), JNK signal transduction and activation, mitotic cells cycle, so as to participate in the pathogenesis of psoriasis. At the same time the ZhuHuang granules miRNA expression of the intervention, and the results show the miR-192, miR-194, miR-3175, miR-18a, miR-629, miR-21, miR-25, miR-100, miR-199a-5p, miR-99a, miR-409-3p miR-125-b, miR-99-b miRNA have significant differences such as two-way of regulating functions.Conclusion:1.67differenct miRNAs such as miR-31involved in the incidence of psoriasis and played a very important role;2. The regulation of genes miRNA differences may be affected by the MAPK signals of pathways that participate in the pathogenesis of psoriasis and prognosis, in cell proliferation, inflammation formation, the growth of blood vessels to constitute the influence, etc;3. Five specific miRNAs such as miR-31is expected to become potential markers of psoriasis early diagnosis and therapeutic targets;4. The effect of treating psoriasis with Chinese medicine Zhu-Huang granules is distinct. The granule have the two-way adjustment function to different miRNAs such as miR-192. The research provide the objective basis for treatment of psoriasis with traditional Chinese medicine;5. The results provide clues for the next step for psoriasis miRNA target genes relating to function and molecular markers.