Diabetes, Metabolic Syndrome and Obesity (May 2021)
A-FABP-PTEN/AKT Regulates Insulin Resistance in Preadipocyte Cell 3T3-L1 Cells
Abstract
Rensiqin Wu,1 Hui Wang,1 Jian Huangfu,1 Rui Xiao2 1Department of Endocrinology, The Affiliated Hospital of Inner Mongolia Medical University, Hohhot, 010000, Inner Mongolia, People’s Republic of China; 2Key Laboratory of Molecular Pathology, Inner Mongolia Medical University, Huhhot, 010000, Inner Mongolia, People’s Republic of ChinaCorrespondence: Jian HuangfuDepartment of Endocrinology, The Affiliated Hospital of Inner Mongolia Medical University, NO.1st Tunnel North Road, Hohhot, 010000, Inner Mongolia, People’s Republic of ChinaTel +86-18686053304Email [email protected] XiaoKey Laboratory of Molecular Pathology, Inner Mongolia Medical University, NO.1st Tunnel North Road, Huhhot, 010000, Inner Mongolia, People’s Republic of ChinaTel +86-18686053304Email [email protected]: The purpose of this study was to explore the regulation of A-FABP-PTEN/AKT on insulin resistance in preadipocyte 3T3-L1 cell.Methods: siRNA interference method was used to knock-down the A-FABP expression in 3T3-L1 cells. The cell proliferation was detected by oil-O staining and MTT. The protein and mRNA expression levels of A-FABP, PTEN and AKT were detected by Western blot and qPCR.Results: Inhibition of A-FABP expression increased cell proliferation activity of the 3T3-L1 cells. Moreover, siRNA3 significantly reduced A-FABP mRNA expression compared with siRNA1 and siRNA2 (P< 0.05). The A-FABP mRNA level was significantly increased in the induced 3T3-L1 cells, while the PTEN mRNA expression was significantly decreased (P< 0.05). Inhibition of A-FABP can significantly increase the PTEN mRNA expression in the process of induced 3T3-L1 cells (P< 0.05). Overexpression of A-FABP can also increase the PTEN mRNA expression in the process of 3T3-L1 cell proliferation (P< 0.05). Furthermore, the protein expression levels of PTEN and p-AKT expression were not changed in the process of 3T3-L1 cell proliferation with or without A-FABP interference (P> 0.05). However, inhibition of A-FABP significantly increased the PTEN protein expression and reduced the p-AKT protein expression in the induced 3T3-L1 cells.Conclusion: Our finding suggested that A-FABP can directly inhibit the phosphorylation of AKT and increase the PTEN expression in the process of normal adipocyte differentiation, which speculated that A-FABP played a crucial role by adjusting the AKT activity in the process of adipocyte differentiation.Keywords: A-FABP, PTEN, AKT, preadipocyte differentiation, insulin resistance
