Significance Of ERα/TGF-β1/Smad Signaling Pathway In Transdifferentiation Of Peritoneal Mesothelial Cells

Part ⅠThe Correlation between ERα and Peritoneal Fibrosis in the Peritoneal Dialysis Effluent of Female Peritoneal Dialysis PatientsObjective:To investigate the difference between ERα and fibrosis markers in the waste dialysate of female peritoneal dialysis patients,and also discuss the Correlation between ERα and peritoneal fibrosis.Methods:To collect 40 cases clinical datas of followed-up female patients who accept continuous ambulatory peritoneal dialysis in May 2016 and May 2017 in our hospital,which including name,age,primary disease,duration of dialysis,complications,occurrence of peritonitis,blood pressure and nutritional status.These patients were treated with 2.5%glucosed-based dialysate and left liquid in the abdominal cavity overnight for the day before the study.10ml waste dialysate was collected in the next morning.Patients were divided into two groups according to whether they were postmenopausal.The content of ERα,TGF-β1,collagen Ⅰ,collagen Ⅲ,α-SMA,FN and E-Cadherin in the peritoneal dialysis effluent were determined by enzyme-linked immunosorbent assay(ELISA).Results:(1)The content of ERα was higher in the postmenopausal women’s waste dialysate than that in the premenopausal women’s waste dialysate,the difference was statistically significant.(2)The content of COL-Ⅰ,COL-Ⅲ were higher in the postmenopausal women’s waste dialysate than that in the premenopausal women’s waste dialysate,the difference were statistically significant.But the the content of TGF-β1,α-SMA,FN and E-Cadherin in the waste dialysate of two groups had no difference.(3)In the waste dialysate of premenopausal women,ERα and TGF-β1,COL-Ⅰ,COL-Ⅲ were positively correlated,but there were no correlation between ERα and α-SMA,FN,E-Cadherin.In the waste dialysate of postmenopausal women,ERα and TGF-β1,COL-1,COL-Ⅲ were also positively correlated,but there were no correlation between ERα and α-SMA,FN,E-Cadherin.Conclusion:ERα in the peritoneal dialysis effluent of female peritoneal dialysis patients was correlated with parts of the fibrosis markers,which may indicate that ERα may assist in the occurrence and development of peritoneal fibrosis.Part ⅡThe Significance of ERα/TGF-β1/Smad Signaling Pathway in the EMT of C57BL/6 Mice Peritoneal Mesothelial CellsObjective:To establish EMT model by high glucose inducing C57BL/6 mice peritoneal mesothelial cells,measure the protein expression of ERα、TGF-β1、Smad2/3 of ERα/TGF-β1/Smad Signaling Pathway and EMT related protein such as E-cadherin、Vimentin,and initially investigate the Significance of ERα/TGF-β1/Smad signaling pathway in the EMT of C57BL/6 mice peritoneal mesothelial cells.Methods:By digesting C57BL/6 mice peritoneal mesothelial cells with trypsin,after subculture and evaluation,collect the second generation cells for the next experiment.Dividing these cells into different groups:(1)control group;(2)the high glucose group(HG);(3)the ERα agonist group(PPT);(4)the ERα inhibitor group(TAM);(5)ERα agonist and Smad3 agonist group(PPT+SIS).ERα、TGF-β1、Smad2/3 and EMT related mark proteins of C57BL/6 mice peritoneal mesothelial cells were evaluated by immunofluorescence and Western Blot.Results:(1)The immunofluorescence and Western Blot evaluation pointed that,the protein expression of ERα increased after affected by high glucose;(2)Interposed by different drugs,The immunofluorescence and Western Blot evaluation indicated that the protein expression of E-cadherin decreased in the order of Control group、TAM group、PPT+SIS3 group、HG group、PPT group;the protein expression of Vimentin increased in the order of Control group、TAM group、PPT+SIS3 group、HG group、PPT group;the protein expression of TGF-β1 increased in the order of Control group、TAM group、PPT+SIS3 group、HG group、PPT group;the protein expression of Smad2/3 increased in the order of Control group、TAM group、PPT+SIS3 group、HG group、PPT group.All the differences were statistically significant(P<0.05).Conclusion:High glucose can induce the EMT process of C57BL/6 mice peritoneal mesothelial cells,which leads to the abnormal protein expression of ERα/TGF-β1/Smad signaling pathway;The ERα inhibitor TAM can suppress the occurrence of EMT by reducing the protein expression which related with TGF-β1/Smad signaling pathway.