Long Noncoding RNA Expression Differences Studies In Patients With Non-IgA Mesangial Proliferative Glomerulonephritis

Background:Mesangial proliferative glomerulonephritis (MsPGN) is one of autoimmune-mediated primary glomerulonephritis. It is classified as type Ⅱ lupus nephritis by the International Society of Nephrology and the Renal Pathology Society, but the mechanism underlying this disease remains largely unclear. According to the immune pathogenesis, MsPGN can be divided into IgA MsPGN (immune deposits are mainly to IgA in mesangial area) and non-IgA MsPGN (IgA deposition in the glomerular mesangial area is not observed).Long non-coding RNAs (lncRNAs) are a form of noncoding RNA and stranscripts longer than200nucleotides, but little or no protein-coding capacity. In the past, people had thought that lncRNA is by-products in the process of transcription and no function. Recent research shows that lncRNAs have important regulatory functions in gene transcription, post-transcriptional mRNA controls and epigenetic modifications. The discovery of these functions to make scientific researcher have focused on lncRNAs potential role in the pathogenesis of disease, and there have been many new findings.Purpose:Through microarray analysis differentially expressed mRNA and lncRNA profile between patients with non-IgA MsPGN and the controls, then screened significant differences expression lncRNA and coordinate changed gene that in genomic context of lncRNAs, thus to explore lncRNA potential role in the pathogenesis of non-IgA MsPGN.Methods:Collect4renal cortical tissues from patients with non-IgA MsPGN and a normal renal cortical tissue. Total RNA was extracted using trizol reagent. Total RNA from each sample was quantified using the NanoDrop ND-1000and RNA integrity was assessed using standard denaturing agarose gel electrophoresis. Total RNA was prepared to ds-cDNA through reverse transcription with by Invitrogen Superscript ds-cDNA synthesis and ds-cDNA labeling with NimbleGen one-color DNA labeling kit, then do array hybridization using Human12xl35K lncRNA expression array; Array scanning using the Axon GenePix4000B microarray scanner. Scanned images were then imported into NimbleScan software and Agilent GeneSpring GX software for grid alignment and expression data analysis, and generated the experimental results data. These datas through GO analysis, Pathway analysis and the gene loci correlation analysis of mRNA and lncRNA were applied to find out lncRNA that are closely related to non-IgA MsPGN. Finally, the RT-PCR was performed to test part of results of array test results, to verify the reliability of array test results.Results:Through Fold Change filtering, made data to achieve significant level to identify differentially expressed mRNAs and lncRNA (P<0.05). There are3095(-16.42%) mRNAs (fold-change≧4) differential expression of which1461up-regulated and1634down-regulated. There are2846(-15.37%) lncRNAs (fold-change≧3) differential expression of which1431up-regulated and1415down-regulated. Through GO analysis, Pathway analysis and the gene loci correlation analysis of mRNA and lncRNA, we found that5lncRNA potential role in the pathogenesis of non-IgA MsPGN:AF1180924, AK092233, AK130579, AK023598, and AK055915.Conclusion:The patients with non-IgA MsPGN were immune function disorders and cell apoptosis self of signal transmission and inflammation of the receptor activity were dysregulations. LncRNA can potential regulation these genes that association with these functions, so I think that lncRNA may play an important role in the happening of non-IgA MsPGN and the development process.