The Mechanisms Of Twist To Promote The Peritoneal Fibrosis Of Human Peritoneal Mesothelial Cells In Peritoneal Dialysis Patients

ESRD is a common and serious disease, at an annual growth rate of10%in recentyears. Peritoneal dialysis (PD) is one method of therapy for end stage renal disease(ESRD), it use the peritoneum as a semipermeable membrane to ultrafiltrationanddiffusion. Long-term PD causes functional and structural alterations of the peritonealmembrane, so patients must be changed another treatment strategy.Epithelial-mesenchymal transition (EMT) of peritoneal mesothelial cells is considered asthe beginning process of peritoneal fibrosis which is related to peritoneal early damage.Myofibroblasts and extracellular matrix (ECM) under mesothelial cells is the importantchanges of peritoneal fibrosis. EMT is a necessary process of tumor development,chronic inflammation and fibrosis. Twist play an important role in EMT. Our previousstudy confirmed that overexpression of Twist promotes epithelial-to-mesenchymaltransition of Human peritoneal mesothelial cells. However, previous studies mainly invitro. Twist whether to promote EMT In vivo and through which the relevant molecularregulation is unclear. The mechanisms and regulatory molecules of EMT play a crucialrole to understand and treatment of peritoneal fibrosis.【Objectives】To explore the mechanisms of Twist on the peritoneal fibrosis of human peritoneal mesothelial cells (HPMCs) in peritoneal dialysis (PD) patients.【Methods】(1)The HPMCs from peritoneal dialysis fluid (in PD patients) were collected andcultured. And the expression and subcellular location of E-cadherin, α-SMA, Twist andBmi-1, YB-1were detected by immunofluorescence cytochemistry technique andWestern-blot.(2) Twist, Bmi-1,YB-1, E-cadherin and α-SMA were detected by immunofluorescencecytochemistry technique and Western blotting after being transfected the plasmidspcDNA3.1-twist and empty vector pcDNA3.1into HPMCs with Lipofectamine2000invitro.(3) Twist, Bmi-1, YB-1, E-cadherin and α-SMA were detected by Western blotting andimmunofluorescence cytochemistry technique after silencing the expression Twist bysiRNAand empty vector pSlience.(4)Bmi-1, E-cadherin and α-SMA were detected by Western blotting andimmunofluorescence cytochemistry technique after silencing the expression Bmi-1bysiRNAand empty vector pSlience.【Results】(1) The expression of E-cadherin were gradually decreased and expression of Twist,α-SMA and Bmi-1， YB-1were increased along with PD time increase byimmunofluorescence cytochemistry technique and Western-blot.(2) Compared with empty vector pcDNA3.1into HPMCs, the expression of E-cadherinwere significantly reduced and expression of α-SMA, Twist, Bmi-1， YB-1weresignificantly increased (P<0.05) after transfected the plasmids pcDNA3.1-twist.(3) The expression of E-cadherin were significantly increased and the expression ofα-SMA, Twist, Bmi-1，YB-1were significantly deceased after silencing the expressiontwist by siRNA.(3) The expression of E-cadherin were significantly increased and the expression ofα-SMA, Bmi-1were significantly deceased after silencing the expression Bmi-1by siRNA.【Conclusion】Twist takes part in the progression of peritoneal fibrosis, and it cans promotesepithelial-to-mesenchymal transition of human peritoneal mesothelial cells throughBmi-1, YB-1expression.