The Role Of PTEN-mediated PI3K/Akt Signaling Pathway In Podocyte Injury

Objective:To study the role of Akt and its downstream molecule GSK3β in the PTEN-PI3K/Akt signaling pathway in glomerular podocyte injury,and to clarify the molecular mechanism of PTEN-mediated PI3K/Akt signaling pathway in the regulation of podocyte injury.Methods:The podocytes was cultured in vitro.The experiment was divided into two parts.The first part was divided into control group,puromycin aminonucleotide group(PAN group),DEX group(PAN+DEX group).The second part was divided into ①Empty cell group;②Empty vector group;③OE-PTEN group;④normal control group(Contron group);⑤si-PTEN group.The control was treated with 0.02%DMSO RPMI 1640 medium to culture glomerular podocytes,the puromycin aminonucleotide group was treated with Puromycin aminonucleotide(final concentration 50μg/ml),and the DEX+PAN group:DEX 1 μmol/L and PAN(final concentration of 50μg/ml)was added to culture glomerular podocytes.The PTEN silencing group and the PTEN overexpression group were carried out according to the instructions of the kit and the operating procedures of this experiment.The changes of the glomerular podocyte cell body area in each group were observed by inverted phase contrast microscope;the expression and distribution of related proteins were detected by laser confocal respectively;Flow cytometry was used to detect and analyze the changes in the apoptosis rate of glomerular podocytes in different group;Western blotting technology was used to detect the expression of glomerular podocyte-related proteins;transmission electron microscopy(TEM)was used to observe the morphology and mitochondrial structure of glomerular podocytes in different group.Results:①After PAN-injured podocytes,the expression of PTEN protein decreased,the apoptosis rate increased,and autophagy was inhibited,and DEX intervention could reverse the above changes;after PAN-injured podocytes,the expressions of p-Akt and p-GSK3 β decreased,and DEX could reverse podocytes The expression of p-Akt and p-GSK3 β in cells;compared with the Control,after silencing PTEN gene,mitochondria were gradually swollen and rounded,mitochondrial cristae were disordered,mitophagy was inhibited,the expression of p-Akt was increased,and the overexpression of p-Akt was increased.These changes can be reversed after PTEN gene.Conclusion:In this study,We found that dexamethasone may upregulate the expression of PTEN and inhibit the PI3K/Akt signaling pathway to protect the effect of puromycin aminonucleotide-induced glomerular podocyte injury;PTEN can regulate glomerular podocyte injury by inhibiting the PI3K/AKT signaling pathway and activating autophagy.