Background <p>MicroRNAs (miRNAs) are post-transcriptional regulators that influence gene expression and dysregulation has been linked to the pathogenesis of ulcerative colitis (UC). The current study investigated the regulatory axis between hsa-mir-10a-5p and RNF186 in UC inflammation.</p> Methods <p>RNF186 was identified as a target of hsa-mir-10a-5p by bioinformatics prediction and dual-luciferase reporter assay. A UC cell model was established by lipopolysaccharide (LPS) stimulation of HT-29 intestinal epithelial cells. Gain-of-function (overexpression) and loss-of-function (knockdown) experiments targeting hsa-mir-10a-5p and RNF186 were performed to assess function and mRNA and protein expression were measured by quantitative real-time PCR (qRT-PCR) and western blotting. Apoptosis was quantified by flow cytometry.</p> Results <p>hsa-mir-10a-5p was shown to bind to the 3’UTR of RNF186 by dual-luciferase assay and a negative correlation shown between the expression of hsa-mir-10a-5p and RNF186 in a UC model. hsa-mir-10a-5p overexpression downregulated RNF186 expression, reduced pro-inflammatory cytokines, TNF-α, IL-6, and attenuated apoptosis. hsa-mir-10a-5p knockdown upregulated RNF186 expression and exacerbated inflammatory responses. RNF186 knockdown produced a similar anti-inflammatory effect to that observed with hsa-mir-10a-5p overexpression. RNF186 overexpression increased inflammatory damage.</p> Conclusion <p>hsa-mir-10a-5p negatively regulated RNF186 and modulated the inflammation characteristic of UC. The hsa-mir-10a-5p/RNF186 axis may be a novel therapeutic target for UC.</p>