The Regulation Function Of Wnt/?-catenin Signal Pathway On The Injury Of Pulmonary Microvascular Endothelial Cells Induced By Lipopolysaccharide

Objectives: The purpose of this study is to investigate effect and molecular biological mechanisms of the wnt signaling pathway in the injury of pulmonary microvascular endothelial cells induced by lipopolysaccharide.Methods: Firstly,an improved tissue explants adherent method was used to isolate and culture the Pulmonary Microvascular Endothelial Cells(PMVECs)from SD rat,and the cells were identified by immunocytochemistry.Secondly,cells were randomly divided into four groups: ? control group: We treated cells with certain volume of serum free medium.? XAV939 group: Cells were treated with XAV939 of which concentration is 10ng/ml.? LPS group: Cells were treated with lipopolysaccharide(LPS)of which concentration is 10?g/ml.? LPS+XAV939 group: we treated cells with mixed liquor of LPS and XAV939.When other conditions are same,all cells were processed 12 hours.Finally,the amount of apoptotic cells was measured by flow cytometry.We detected the protein expression level of the?-catenin,Cyclin D proteins related the wnt signaling pathway and TNF-? in the cells using western blotting.And we also detected genetic level change of?-catenin related the wnt signaling pathway using Real time PCR.Results: 1.The monolayer of primary PMVECs displayed a typical cobble stone-like appearance under the microscope.The cells were positive for VIII factor related antigen using immunocytochemistry.2.The result of FCM indicated that LPS-induced cells apoptosis showed time dependence.And XAV939 could not directly induce apoptosis(P<0.05).3.In the different treatment groups,LPS-induced cells apoptosis was significantly higher than the control group(P<0.05).The?-catenin?Cyclind and TNF-?protein levels and ?-catenin gene expression levels of LPS group cells were significant higher as compared with control group(**P<0.05),and the protein and gene expression levels of LPS+XAV939 group were significant decreased than that of the control group(*P<0.05).Conclusion: In this project,we found that wnt/?-catenin signaling pathway inhibition could effectively decreased the injury of PMVECs.That is to say,wnt signaling pathway participated in the injure process of pulmonary microvascular endothelial cells induced by LPS.