Abstract
Objective To investigate the effect of glimepiride on PI3K/Akt pathway in rat mandible osteoblasts.Methods Primary osteoblasts were isolated and cultured.Osteoblasts were seeded onto 25 cm2 plates at a density of 1× 105 cells/plate.All the groups were cultured for 15 min,30min,60 min and 120 min,respectively.The cell lyses were collected equally to ensure each sample containing equal protein.The electrophoresis was performed and detected with antibodies against PI3- kinase P85,IRS-1,P-IRS-1,Akt,and P-Akt (Ser-473),which were diluted in TBS-T for 4 hours at room temperature.After three washes by TBS-T,the blots were subsequently incubated with a sheep anti-rabbit peroxidase-conjugated secondary antibody for 50 min at room temperature.The blots were visualized by enhanced chemiluminescence using Labworks 4.5 Image Acquisition System.For normalization purposes,the same blot was also probed with rabbit anti-rat β-action antibody.Results The expression of PI3K P85,Akt/P Akt and IRS-1/P-IRS-1 protein expression was significantly increased by glimepiride in the cultured osteoblasts.Conclusions Glimepiride activates the PI3K/Akt pathway in osteoblasts. Key words: Glimepiride; Osteoblasts; Mandible
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