Ifferential Expressions Of Specific Serum Micrornas In Patients With Nephrotic Syndrome

Nephrotic syndrome (NS) is a syndrome but not a single disease which is characterized by massive proteinuria, which usually leads to hypoproteinemia, triglicerides, edema, hyperlipidemia with elevated cholesterols and other lipids. Nephrotic syndrome can be caused by one or more different diseases.Because the permeability of glomerular basement membrane increases, massive plasma proteins are lost. The most frenquent complications of nephrotic syndrome are thrombosis, embolism, acute renal failure, protein metabolism, fat metabolism, renal tubule hypofunction and endocrine disturbance. Nephrotic syndrome can generally divided into two kinds:the primarily nephrotic syndrome and secondary nephrotic syndrome. At present, renal biopsy is the most common diagnosis means of nephrotic syndrome, but it’s invasive and insensitive. Thus, it is important to develop new sensitive specific and significant means for early identification of etiopathogenisis and pathology type with nephrotic syndrome.Nowadays,serum become one of the best origin of research for its easy collection and better record of physiology condition. Also,miRNAs have become the hot pot in our research since the first two miRNAs (lin-4,let-7) were discovered.Therefore, the study on miRNAs in NS can provide new insights into the diagnosis of different pathology type of nephrotic syndrome and the pathogenesis.Objective:Using the real-time quantitative PCR to explore that whether there were differential expressions of miR-181a、miR-483-5p and miR-557in the serum between different pathology type patients with nephrotic syndrome and healthy controls. To explore whether the differential expressions of miRNAs can provide new ways into the diagnosis of different pathology type of nephrotic syndrome and the pathogenesis.Methods:Serum samples were taken from40NS patients from nephrology department of181st hospital, Guangxi, China, of whom10were diadonsed with MsPGN,10were diadonsed with MCNS,10were diadonsed with MN,and10were diadonsed with FSGS. The diagnosis was confirmed by renal biopsy and clinical evidence.16samples of healthy people were taken from the physical center of People’s Liberation Army the181st hospital. All samples were collected only after receiving informed consent from the subject. The potential NS-related miR-181a、 miR-483-5p and miR-557were chosen according to the mirbase (http://www.mirbase.org) previously, the gene orders were find in GeneBank, then the primers and the probes were designed using a software named Primer-Express2.0. Total RNA was extracted from serum using Trizol (Invitrogen). The concentration and quality of total RNA were measured by the UV absorbance at260nm and280nm (A260/280), then miRNA quantitative RT-PCR (qRT-PCR) analysis system (ABI PRISM(?)7500Sequence Detection System) was used to quantify miR-181a、miR-483-5p、miR-557expression.Constructing the real-time quantitative PCR system of miR-181a、miR-483-5p and miR-557,then analyze the datas using the statistical softwares.Results:According to normalization of the raw date, then we found that these three miRNAs were diffe rentially expressed; in which miR-483-5p and miR-557were down-regulated and miR-181a was up-regulated between NS patients and healthy cont rols. Then, we compared the expression of miRNAs between the different pathology type of NS pool and the healthy controls pool, between each two different pathology types of NS pools, respectively.We find significant differential expressions,too. In the GSPS group, miR-181a were detected at higher level in the blood, but miR-557showed a trend toward significantly reduced level.Conclusions:Realtime-PCR was an effective approach for detecting expression level of the miRNA. The expression of miRNAs between the different pathology type of NS pool and the healthy controls pool, between each two different pathology types of NS pools,have differential expressions, miR-181a was up-regulated between NS patients and healthy cont rols.The differential expressions of serum microRNAs in patients with Nephrotic Syndrome providing people a new point of view to better understand the pathogenesis of NS and early diagnose in the diagnosis of different pathology type of nephrotic syndrome.