The Role And Mechanism Of MiR-939 Blocker In The Treatment Of Nephrotic Syndrome By Regulating CD2AP

Objective:The injury of glomerular podocytes is a key factor in the production of proteinuria and is the main mechanism of the pathogenesis of nephrotic syndrome.CD2-associated protein(CD2AP)is the most important narrowing factor for maintaining podocyte integrity and reducing proteinuria.One of the septum proteins.Defects in CD2 AP or abnormal transcription can lead to podocyte failure and proteinuria glomerular disease.There are many different small RNAs(miRNAs)in patients with nephrotic syndrome,which may be related to the occurrence of nephrotic syndrome.The structure and function of various cells in the kidney,including podocytes,are regulated by miRNAs.Abnormal expression of some miRNAs leads to decreased podocyte fusion and CDAP expression.The main objective of this study was to explore the discovery of specific miR-939 that regulates its expression by targeting the CD2 AP promoter region,thereby generating a new horizon for the pathogenesis of podocyte injury.The therapeutic effect of miR-939 antagonist on nephrotic syndrome was observed by establishing an animal model.Methods:RegRNA 2.0 online software predicts that the CD2 AP promoter region has binding sites of miR-939,while the 3,UTR region of CD2 AP has no miR-939 binding site;Fluorescence in situ hybridization(FISH)detects position of 939 in HEK293 cells,PCR method was used to obtain the 5’ flanking sequence of CD2 AP gene and a series of 5’-end deletions,which were inserted into pGL3-Basic vector.The double luciferase reporter detection system was used to identify its promoter activity.Using the point mutation/deletion mutation technique to identify whether miR-939 can target to the binding site to regulates CD2 AP promoter activity after transfection of miR-939;Chromatin immunoprecipitation(ChIP)detects the enrichment of RNA polymerase II in the human CD2 A promoter region;in peripheral blood of patients with nephrotic syndrome,expression levels of miR-939 and CD2 AP were detected;Establishment of Adriamycin-induced rat nephrotic syndrome model,intravenous injection of miR-939 blocker,through Observeing renal appearance,renal pathology,blood biochemistry and CD2 AP expression to detect the therapeutic effect of miR-939 for nephrotic syndrome.Results:Bioinformatics predicted that there are two miR-939 targeting binding sites in the CD2 AP promoter region;FISH confirmed that miR-939 can be found in the nucleus of HEK293 cells;miR-939 binds to the 491--468 binding site by binding to the CD2 AP promoter.Negative regulation of CD2AP;ChIP assay after transfection of miR-939 revealed a decrease in enrichment of RNA polymerase II in the CD2 AP promoter region.In the peripheral blood of patients with nephrotic syndrome,the expression of miR-939 is increased and the expression of CD2 AP is decreased.miR-939 blocker can improve the appearance of kidney in the kidney disease model,blood biochemical indicators,reduce renal tissue lesions,increase CD2 AP expression.Conclusion:CD2AP is a target gene of miR-939.In the nucleus,miR-939 down-regulates the transcription and expression of CD2 AP by reducing the enrichment of RNA polymerase II in the promoter region.miR-939 blocker has certain treatment for nephrotic syndrome.