| Objective:The effects of microRNA-203a-5p on cytokines such as IL-6,TNF-? and IL-1? in HaCaT cells were analyzed by over expression or inhibition of microRNA-203a-5p expression in cell experiments,and the targeting effects of microRNA-203a-5p on these cytokines were identified.Furthermore,the expression of microRNA-203a-5p,IL-6,TNF-? and IL-1? in the skin of patients with atopic dermatitis was combined to further verify the relationship between them,and to analyze their role in the pathogenesis of atopic dermatitis,providing a theoretical basis for targeted therapy with microRNA-203a-5p and its downstream target genes,and providing a new idea for the treatment of atopic dermatitis.Methods:1.HaCaT cell lines were cultured in vitro and transfected into four groups: mimics group,mimics NC group,inhibitor group and inhibitor NC group.After transfection,the expression levels of microRNA-203a-5p and IL-6,TNF-? and IL-1? were further detected by qPCR.2.The total RNA of the two groups was extracted by Trizol method after quick-frozen liquid nitrogen grinding.The expression levels of microRNA-203a-5p,IL-6,TNF-? and IL-1? in the two groups were detected by qPCR.Results:1.In cell experiment,HaCat cells were transfected with four groups of samples: mimic group,inhibitor group and negative control group.The results showed that the expression level of microRNA-203a-5p mimics was significantly higher than that of the negative control group(P < 0.01),which proved that the transfection was successful.The expression levels of TNF-? and IL-6 in HaCat cells were significantly lower than those in the control group(P < 0.01),while the expression levels of IL-1? increased(P < 0.05).After the transfection of micro RNA-203a-5p inhibitor,the expression of TNF-? increased and the expression of IL-6 and IL-1? decreased in HaCat cells(P< 0.05).2.The results of q-PCR showed that the expression of microRNA-203a-5p,IL-6 and TNF-? in skin scraps of atopic dermatitis patients was higher than that of normal control group(P < 0.01),and the expression of IL-1? was slightly higher than that of normal control group(P > 0.05).Spearman correlation analysis showed that the expression of microRNA-203a-5p and TNF-? was not significantly different(P > 0.05).There was no correlation between the expression of IL-6,IL-1?(r 0.26,0.15,0.18,P > 0.05).Conclusion:(1)The increased expression of microRNA-203a-5p,TNF-? IL-6 and IL-1? in the dandruff of AD patients suggests that microRNA-203a-5p,TNF-?,IL-6 and IL-1? are involved in the occurrence and development of AD.(2)MiR-203a-5p in HaCat cells can inhibit the expression of TNF-? and IL-6 and promote the expression of IL-1?.However,in the study of AD patients' dandruff,the abnormal expression of TNF-?,IL-6 and IL-1? increased,and no direct and effective regulation of TNF-?,IL-6 and IL-1? by mir-203a-5P was found.Other regulation methods are unknown.It is speculated that the reason may be due to the dysfunction of epidermal barrier or the remodeling of epidermis in atopic dermatitis.During the inflammatory reaction stage,the regulatory role of microRNA-203a-5P is changed or inhibited.(3)In this study,the targeting regulatory effects of microRNA-203a-5P on TNF-?,IL-6 and IL-1? were verified at the cellular level,and the possible pathogenesis and regulatory pathways of microRNA-203a-5P in the occurrence and development of AD were preliminarily explored in combination with the expression of microRNA-203a-5P in AD dandruff tissues and normal skin tissues.The regulation of microRNA-203a-5P on TNF-?,IL-6 and IL-1? at the gene level was also studied.Controlling effect,revealing the possible pathogenesis of AD from the perspective of microRNAs-pathways-inflammatory factors-diseases,but there is no in-depth study on the regulation function of specific microRNAs and their related susceptible genes,cytokines and so on.Next,we will further study their expression and interaction at the protein level,and the molecular mechanism of microRNA-203a-5P in the pathogenesis of AD.The cellular pathways involved were further studied,and the role of microRNA-203a-5P as a target for the diagnosis,treatment and prognosis analysis of AD and the whole network regulation system in the pathogenesis and evolution of AD was discussed,which provided a theoretical basis for the targeting therapy of AD and the design of effective inhibitors in the future. |
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