The Experimental Study Of Conditioned Medium From Human Umbilical Cord Mesenchymal Stem Cells And Zinc On EMT In Human Peritoneal Mesothelial Cells Induced By High Glucose

Objective:（A）To explore the effects of conditioned medium（CM）from human umbilical cord mesenchymal stem cells（hUC-MSCs）on high glucose（HG）-induced EMT in human peritoneal mesothelium cells（HPMCs）and its underlying mechanism.（B）To explore the effects of Zinc on HG induced EMT in HPMCs and its underlying mechanism.Methods:（A）The HPMCs were divided into three groups:（1）control group,（2）HG group,（3）CM group.The effect of HG and MSC-CM on proliferation in HPMCs was detected by CCK-8.The morphology changes was observed under microscope.The expression of E-cadherin,Vimentin,α-SMA andβ-catenin in HPMCs was detected by Western Blot.（B）The HPMCs were divided into four groups:（1）control group,（2）HG group,（3）HG+TPEN group,（4）HG+Zn group.The expression of TGF-β₁ in HPMCs was detected by ELISA.The expression of E-cadherin,Vimentin,p-Smad2/3 and p-Smad7 was detected by Wsetern Blot.Results:（A）（1）HG reduced the proliferation of HPMCs,and MSC-CM promoted the proliferation of HPMCs.（2）Compared to the control group,HPMCs in the HG group were rare and mostly spindle sharp.And MSC-CM could alleviate the morphological changes of HPMCs above.（3）Compared to the control group,HG treatment reduced the expression levels of E-cadherin,and increased the expression levels of Vimentin andα-SMA.However,MSC-CM up-regulated the expression of E-cadherin,and down-regulated the expression of Vimentin andα-SMA.（4）Meanwhile,HG increased the expression ofβ-catenin in HPMCs.MSC-CM reduced the expression ofβ-catenin compared with the HG group.（B）（1）Compared to the control group,HG treatment reduced the expression levels of E-cadherin,and increased the expression levels of Vimentin.Additionally,TPEN treatment enhanced the effects of HG on HPMCs.However,Zinc supplementation significantly up-regulated the expression levels of E-cadherin,and down-regulated the expression of Vimentin.（2）Meanwhile,HG increased TGF-β₁ and p-Smad2/3,and reduced p-Smad7.Zinc supplementation reduced TGF-β₁ and p-Smad2/3,and increased p-Smad7 compared with the HG group.Conclusions:（A）Conditioned medium from human umbilical cord mesenchymal stem cells attenuate high glucose-induced EMT by down-regulating Wnt/β-catenin pathway in human peritoneal mesothelial cells.（B）Zinc supplementation attenuates the HG-reduced EMT in HPMCs by down-regulating TGF-β/Smad pathway.