Role Of PPARγ In High Glucose-induced Insulin Resistance And Susceptibility In Vascular Endothelial Cells (HUVECs) By RNAi

Objective:The recombinant vector for shRNA of PPARy gene was constructed and transfected into human umbilical vein endothelial cells (HUVECs), to study the role of PPARy in high glucose-induced insulin resistance(IR) and susceptibility in (HUVECs).Methods:The vector of shRNAPPARy3was chosen and its silencing efficiency is63.2%. Four groups were individed,namely Control group, insulin resistance group, Non-silencing shRNA group, shRNAPPARy3group. First of all,we establish the model of high glucose-induced insulin resistance in HUVECs,and then transfect the combinant plasmid shRNAPPARy3into HUVECs by liposome transien. According the expression levels of PPARy and eNOS, the levels of NO and Ang II to study the role of PPARy in high glucose-induced insulin resistance in HUVECs by RNAi. In addition, we transfect the combinant plasmid shRNAPPARy3into HUVECs,then establish insulin resistance model, to study the role of PPARy in high glucose-induced insulin resistance and susceptibility in HUVECs by RNAi. According to the expression levels of IKKα/β, pS176-IKKa/α/β,Ba,eNOS, and the levels of TNF-a, IL-6, sICAM-1, sVACM-1, to study the mechanism of PPARy by RNAi.Results:The results of NO and Ang II:Regardless of before modeling or after modeling, The low expression of PPARy can make the level of NO significantly decline, the level of Ang Ⅱ significantly rise,which showing that the low expression of PPARy increase the susceptibility of HUVECs IR and increase the degree of HUVECs IR.The resules of TNT-,IL-6,sICAM-1and sVACM-1showing that:The low expression of PPARy made the levels of inflammatory factors significantly rice,to increase the response of inflammatory in HUVECs.The result of protein expression: Silencing the gene of PPARy can make the expression of eNOS and IκBα significantly reduce, the expression of IKKα/β has no significant change,the expression of pS176-IKKα/β significantly rice. Conclusion:The low expression of PPARy increase the susceptibility of HUVECs IR or increase the degree of HUVECs IR.The mechanism of PPARy regulation IR and its susceptibility is depend on the pathway of IKKα/βIκBα/NF-κB.