Dissertation > Excellent graduate degree dissertation topics show

The Relationship between microRNA302c Expression and Epithelial to Mesenchymal Transition in Human Peritoneal Mesothelial Cells of Patients with PD

Author: ChenYuSa
Tutor: XiaoPing
School: Central South University
Course: Internal Medicine
Keywords: Peritoneal Dialysis Peritoneal fibrosis Epithelial cells - mesenchymal cell transdifferentiation Connective tissue growth factor microRNA302c
CLC: R692.5
Type: Master's thesis
Year: 2010
Downloads: 114
Quote: 0
Read: Download Dissertation

Abstract

Background: continuous ambulatory peritoneal dialysis (Continuous ambulatory peritoneal dialysis, CAPD) is one of the end-stage renal disease (End-stage renal disease, ESRD) treatment alternatives. Long-term CAPD patients with peritoneal long-term exposure to biological incompatibility dialysate, resulting in peritoneal mesothelial cell exfoliation eventual peritoneal fibrosis and ultrafiltration failure (Ultrafiltration failure UFF), leading patients to quit peritoneal the main reason of dialysis. Peritoneal mesothelial cells (Peritoneal mesothelial cells, PMCs) is considered to be caused by fibrosis cells, the researchers found that the epithelial cells to mesenchymal cell transdifferentiation (Epithelial-to-mesenchymal transition, EMT) may be muscle fibroblasts one of the important sources of recent studies further evidence of EMT is the start of peritoneal fibrosis link, and TGF-β1 signaling pathway play a decisive role in peritoneal fibrosis, connective tissue growth factor (CTGF) is mainly caused by fibers of TGF-β1 of downstream effectors. microRNA302c belongs microRNA302 one of the family members, the analysis found cases of possible microRNA302c in the regulation of the expression of CTGF played a key role, but there is no reported biological model program. Objective: To investigate micorRNA 302c in the regulation of CTGF expression peritoneal mesothelial cells and EMT in the role and molecular mechanisms. Methods: (1) The Second Xiangya Hospital, Central South University Xiangya Hospital peritoneal dialysis center CAPD patients, dialysis time is divided into two groups: ① newly opened the Tube group (A) (2) peritoneal dialysis for more than a year group ( group B), whichever night stay abdominal dialysate isolated and cultured human peritoneal mesothelial cells, cultured fourteen days identified by inverted phase contrast microscopy and immunohistochemistry (Immunohistochemistry, HIS), after primary cells used in the experiment. (2) The difference between the inverted microscope to observe the changes in cell morphology (3) expression using real-time PCR, Taqman fluorescent probe method the detection microRNA302c in the above-mentioned two groups of peritoneal mesothelial cells; (4), respectively, with Westem-blot , Real time PCR observation of CTGF positive for vimentin and zo-1 protein levels and RNA levels of expression; (5) analysis microRNA302c and CTGF expression and vimentin, zo-1 expression correlation. More than 95% of the cells results: (1) identification of peritoneal dialysis effluent cultured human peritoneal mesothelial cells (HPMCs). (2) The difference between the inverted microscope was cobblestone circular, oval-shaped, epithelioid cells, fusion was observed a small fraction of patients with newly opened tube of peritoneal dialysis effluent culture HPMCs morphological peritoneal omentum culture mesothelial cells similar; while most of the mixed epithelioid and spindle of fiber-like cells exist; Group B patients with peritoneal dialysis effluent culture HPMCs spindle fiber-like cells. (3) Real time PCRTaqman fluorescent probe method the microRNA302c expression in peritoneal mesothelial cells, and group B patients with peritoneal dialysis the effluent culture HPMCs expression level was significantly lower than in group A patients (P lt; 0.05). (4) Real time PCR group B patients with peritoneal dialysis effluent culture HPMCs Zo-1 mRNA expression level was significantly lower than in group A patients (P lt; 0.05); HPMCs CTGF group B patients with peritoneal dialysis effluent culture , Vimentin mRNA expression levels were significantly higher in group A patients (P lt; 0.05). Westem-blot Group B patients with peritoneal dialysis effluent culture HPMCs Zo-1 protein levels reduced expression levels than group A (P lt; 0.05); HPMCs CTGF, Vimentin protein group B patients with peritoneal dialysis effluent culture expression levels than the expression level of the new tube patients raised (P lt; 0.05). (5) A and B groups of patients with peritoneal dialysis effluent culture HPMCs two groups of cells microRNA302c expression of CTGF mRNA expression was negatively correlated with the correlation coefficient r = -0.54; microRNA302c expression of Vimentin mRNA expression was negatively correlated with the correlation coefficient r = -0.884; microRNA302c expression and Zo-1 mRNA in expression showed a positive correlation coefficient r = 0.86; CTGF mRNA and Vimentin in mRNA in expression was positively correlated with a correlation coefficient r = 0.83; CTGF mRNA and Zo-1 mRNA in expression was negatively correlated, the correlation coefficient r = -0.72. Were statistically significant (P lt; 0.05) conclusions: (1) the microRNA302c in peritoneal dialysis effluent culture of HPMCs of expression and microRNA302c expression is significantly reduced as the dialysis time prolonged. Of CTGF on peritoneal dialysis for more than one year (2) the culture HPMCs patients with peritoneal dialysis effluent and EMT-associated molecular markers vimentin and zo-1 expression in the newer open tube obvious abnormalities in patients prompted the extension with the dialysis time , the cultured peritoneal dialysis effluent HPMCs in EMT severity; (3) microRNA302c with HPMCs CTGF and abnormal expression of EMT-related genes, suggesting that microRNA302c with PD patients HPMCs EMT, related to the occurrence.

Related Dissertations

CLC: > Medicine, health > Surgery > Urology ( urinary and reproductive system diseases) > Kidney disease > Renal failure
© 2012 www.DissertationTopic.Net Mobile