microRNA-630 regulates underglycosylated IgA1 production in the tonsils by targeting TLR4 in IgA nephropathy. microRNA-630 regulates underglycosylated IgA1 production in the tonsils by targeting TLR4 in IgA nephropathy

IgA nephropathy (IgAN) is the most common primary glomerular disease. The characteristic pathology involves immune complexes formed by the deposition of IgA1 and underglycosylated IgA1 aggregates in the mesangial area, which may be accompanied by the deposition of IgG and/or IgM and complement components. However, the molecular mechanisms of IgAN remain unclear. In the present study, microarray analysis showed that the expression of miR-630 was significantly reduced in palatal tonsils from IgAN patients compared with chronic tonsillitis. Additionally, bioinformatic analysis showed that Toll-like receptor 4 (TLR4) was the predicted target gene of miR-630 and was regulated by miR-630. When miR-630 was overexpressed in palatal tonsil mononuclear cells from IgAN patients, the expression of TLR4 was reduced and the content of IgA1 in the cell culture supernatant was decreased, and the level of galactosylation in the IgA1 hinge region was increased. Moreover, immunohistochemical analysis showed that the expression of TLR4 in IgAN patients was significantly increased and. After knocking down the expression of TLR4, both the concentration of IgA1 and the binding force of IgA1 with broad bean lectin were significantly reduced in IgAN. Furthermore, the mechanism study demonstrated that TLR4 might regulate the expression of IL-1β and IL-8 through NF-κB signaling pathway to modulate the concentration of IgA1 and the glycosylation level of IgA1. This interesting finding may offer new insight into the molecular mechanism of IgAN. Overall design: To identify the differentially expression of microRNAs in tonsil of patients with IgA nephropathy. Tonsillar tissue specimens were obtained from 24 patients with IgA nephropathy (IgAN) whom had been diagnosed by renal biopsy, used as experimental group. The tonsillar tissue specimens which were obtained from20 patients with chronic tonsillitis (CT) lacking renal diseases were used as control group. Chose two specimens randomly from the above two groups respectively, and use the Agilengt Human miRNA microarrays V19 to detect the microRNAs (miRNAs) profile differentially expressed in tonsil of IgAN. The miRNA microarray differential analysis resulted in 29 significantly deregulated miRNAs after applying a fold change threshold >2, either up regulated or down regulated. Among the 29 significantly deregulated miRNAs, 7 miRNAs were up regulated and 22 miRNAs were down regulated. Tonsillar mononuclear cells (TMCs) were separated by density-gradient centrifugation using Lymphocyte Separation Medium, and we performed quantitative real-time PCR (qRT-PCR) to validate 4 deregulated miRNAs showed in the microarray results. The deregulated miRNAs were miR-630, miR-513b, miR-135a-3p, miR-642-3p respectively.