Research And Validation Of Autophagy Related MiR-125b-5p Downstream Target Genes Of Systemic Lupus Erythematosus (SLE)

Objective:To verify the autophagy related miR-125b-5p downstream target genes UVRAG of systemic lupus erythematosus (SLE), we explored UVB irradiation induced the pathogenesis of SLE for the further perspective of autophagy.Methods:Fristly,293T cells were transfected by the chemical synthesis of miR-125b-5p analog miR-mimic and competitive inhibitor of miR-inhibitor (miR-control). Real-time Quantitative PCR (qRT-PCR) detection system and Western-Blotting were employed to test the effect of miR-125b-5p on the transcription and translation of target gene UVRAG Secondly:3’untranslate region(3’UTR)reperter gene method were used to construct the dual luciferase reporter gene plasmids and the mutant plasmids. These plasmids were tansfected into 293T cells with miR-mimic. To vertify the miR-125b-5p’s target gene was UVRAG, the analysis of hluc/hRluc on Dual luciferase Reporter Assays were used.Results:The results of qRT-PCR showed that:the levels of Uvrag mRNA expression in the group of miR-mimic were significant lower than those in the group of miR-inhibitor(F<0.05). The results of Western blotting thowed that the levels Uvrag protein expression in miR-mimic group was slightly lower than those in the miR-inhibitor group, with the differences in the levels of miR-mimic and miR-inhibitor protein between the two groups achieving statistical significance (P<0.01); The results of Dual luciferase Reporter Assays thowed that:the relative Rluc/Luc ratio in miR-mimic group was slightly lower than those in the miR-inhibitor group(P<0.05).Conclusions:UVRAG is considered as a downstream target genes of autophagy related miR-125b-5p of systemic lupus erythematosus (SLE). May be associated with the UVB irradiation induced abnormal activation of autophagy in patients with SLE.