Downregulation Of PTEN Expression In Renal Tubular Epithelial Cells Induced By High-glucose Cultivation Promotes Fibrogenesis And The Mechanism Of Interference With Medicine

Objective: This study aims to observe the influences of high-glucose on the protein and m RNA expression of PTEN and PI3K/Akt pathway in renal tubular epithelial cells, further investigate effects on the expression of PTEN and PI3K/Akt pathway in renal tubular epithelial cells cultured with Rosuvastatin(Ros), and explore the role and possible mechanism of Ros in renal tubular fibrosis pathogenesis. Methods: The renal tubular epithelial cells(NRK52E cells) treated with high-glucose condition in vitro. Immunocytochemistry and Immunoflurescence cytochemistry were used to detectthe expression of CK-18, E-cadherinin and α-SMA to identify NRK-52 E cells. MTT was used to determine the viability of NRK52 E cells with Ros. The cells were randomly divided into normal glucose(NG) groups: treated with normal glucose[DMEM(glucose 5.5 mmol/L) + 2% FBS], high glucose(HG) groups: treated with high glucose [19.5mmol/L D-glucose + DMEM + 2% FBS] and high glucose+Ros(10u M)(HR) groups: treated with high glucose and10 u M Ros, each group was cultured for 2h, 12 h, 24 h and 48 h. Western blotting was used to detect the proteins expression of PTEN, p-Akt(ser473), E-caderin, α-SMA and Col-Ⅳ under different conditions. Real-time PCR was employed to detect PTEN m RNA expression. Results:(1) Immunofluorescence analysis showed that NRE52 E cells was renal tubular epithelial cells.(2)The results of MTT shown that the concentration of 0 u M to 50 u M Ros had no effect on growth of cells, so we choose 10 u M Ros to co-cultivate cells according to references and this results.(3)Compared with those in the normal glucose group, the protein and m RNA expression of PTEN significantly reduced(P<0.05), and gradually decreased with the extension of high glucose cultivate times, also to the protein expression of E-cadherin. Instead, the expression of p-Akt(Ser473), α-SMA and Col-Ⅳ increased remarkably(P<0.05) in renal tubular epithelium after treated with high glucose for 12 hours. Compared with those in the high glucose group, Ros decreased the expression of p-Akt(Ser473),α-SMA and Col-Ⅳ(P<0.05), and upregulated the expression of PTEN protein and m RNA in renal tubular epithelial cells. Conclusion: High glucose reduced the expression of PTEN and activated PI3K/Akt pathway which contributed to renal tubule epithelial cells epithelial-mesenchymal transition(EMT). Ros could inhibit EMT mediated by high glucose, with the mechanism may relate to up-regulating the expression of PTEN which deactivate PI3K/Akt signal pathway.