Role Of PKCδ In The Phosphorylation And Mitochondrial Translocation Of P66Shc In Human Renal Tubular Cells Induced By High Glucose

Objective:To observe the mRNA expressions and protein phosphorylation levels of PKC8and p66Shc in renal tubular epithelial cells (HK-2) with high glucose stimulatation.To study the effect of PKC8on p66Shc phosphorylation and mitochondrial translocation in HK-2cells.Methods:1. Using different concentrations of glucose to stimulate HK-2cells to detect the gene expression and protein phosphorylation levels of PKC8and p66shc by Real Time PCR and Western Blot; Using high concentrations of glucose to stimulate HK-2cells to detect the gene expression and protein phosphorylation levels of PKC8and p66shc at different time points by Real Time PCR and Western Blot.2.(1) To analysis the level of p66Shc protein phosphorylation with or without high glucose treatment under the intervention of PKC8inhibitor by Western blot and immunofluorescence Technical.(2) HK-2cells were treated as above mentioned, and then extract protein from the cytoplasm and mitochondria to observe the p66Shc protein phosphorylation and mitochondrial translocation by Western blot.Results:1. Both mRNA expression and protein phosphorylation levels of PKCδ and p66Shc increased significantly in a dose and time-dependent manner compared with the control group (P<0.05);2. The pharmacological PKCδ inhibitor Rottlerin significantly attenuated p66Shc phosphorylation level of cytoplasm and mitochondria under high glucose environment compared with the control group (#P <0.01).Conclusion: 1. High glucose can stimulate the transcription and phosphorylation of PKCδ and p66Shc in HK-2cells.2.PKCδ can promote the phosphorylation and mitochondrial translocation of p66Shc in HK-2cells with high glucose stimulation.