Preliminary Study On The Role Of TRNA-derived RNA Fragments In Children With Mesangial Proliferative Glomerulonephritis

Objective:Children’s mesangial proliferative glomerulonephritis(MsPGN)is one of the main pathological types of children’s primary glomerulopathy.Therefore,it is urgent to find a target for the role of mesangial proliferative glomerulonephritis in children..The purpose of this study was to reveal the relationship between the transfer RNA fragment and the proliferating mesangial cells,and to preliminary explore the role of the transfer RNA fragment in the proliferation of mesangial cells.Methods:①Read the research literature to understand the different modeling methods for stimulating mesangial cell proliferation,and then select the appropriate modeling method.Cell Counting Kit-8(CCK-8)was used to determine the proliferation of mesangial cells,and the morphological verification and detection of mesangial cell proliferation markers were used to verify the success of the establishment of an model of mesangial proliferative renal disease.②The classification and functional characteristics of tRFs were sorted out,and different tRFs with different mesangial cells before and after abnormal proliferation were obtained by immunoblotting.③Subsequently,tRFs were analyzed after treatment with normal mesangial cells and TGF-β1 by high-throughput sequencing.④Expression in Mesangial Cells.Validated by qPCR technology,the target genes of differentially expressed tRF were predicted,and GO and signal pathway analysis were performed.Results:①The stimulator was a TGF-β1 stimulating factor with a concentration of 2ng/ml TGF-β1,and the mesangial cells were stimulated for 12 hours as a model for the proliferation of mesangial cells in early period.②Abnormally proliferating mesangial cells produce differential tRFs.③Normal mesangial cells and TGF-β1 stimulated total RNA samples extracted from abnormally proliferating mesangial cells were tested for a total of 468 tRFs.It was found that compared with normal mesangial cells,after A total of 320 up-regulated and 148 down-regulated tRFs were abnormally changed in the abnormally proliferating mesangial cells(fold difference>2,and p<0.05).④Verification found that tDR-000064 and tDR-000103 were significantly reduced.These differentially expressed tRF target genes were analyzed for their roles in protein binding,DNA binding,urogenital development,positive regulation of cell proliferation,and renal system development.These gene products are involved in Ras signaling pathway,Hippo signaling pathway and so on.Conclusions:Differential tRFs exist between TGF-β1 and normal mesangial cells;Differential tRFs that play an important role in the proliferation and development of mesangial cells;tRFs through the downstream target genes Regulation of protein binding and DNA binding during the proliferation of mesangial cells.