Wu Rensiqin, Wang Hui, Huangfu Jian, Xiao Rui
Department of Endocrinology, The Affiliated Hospital of Inner Mongolia Medical University, Hohhot, 010000, Inner Mongolia, People's Republic of China.
Key Laboratory of Molecular Pathology, Inner Mongolia Medical University, Huhhot, 010000, Inner Mongolia, People's Republic of China.
Diabetes Metab Syndr Obes. 2021 May 11;14:2103-2110. doi: 10.2147/DMSO.S305872. eCollection 2021.
The purpose of this study was to explore the regulation of A-FABP-PTEN/AKT on insulin resistance in preadipocyte 3T3-L1 cell.
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.
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 (<0.05). The A-FABP mRNA level was significantly increased in the induced 3T3-L1 cells, while the PTEN mRNA expression was significantly decreased (<0.05). Inhibition of A-FABP can significantly increase the PTEN mRNA expression in the process of induced 3T3-L1 cells (<0.05). Overexpression of A-FABP can also increase the PTEN mRNA expression in the process of 3T3-L1 cell proliferation (<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 (>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.
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.
本研究旨在探讨A-FABP-PTEN/AKT对前脂肪细胞3T3-L1细胞胰岛素抵抗的调控作用。
采用siRNA干扰法敲低3T3-L1细胞中A-FABP的表达。通过油红O染色和MTT检测细胞增殖情况。采用蛋白质免疫印迹法(Western blot)和实时定量聚合酶链反应(qPCR)检测A-FABP、PTEN和AKT的蛋白质及mRNA表达水平。
抑制A-FABP表达可增加3T3-L1细胞的增殖活性。此外,与siRNA1和siRNA2相比,siRNA3显著降低了A-FABP mRNA表达(<0.05)。诱导后的3T3-L1细胞中A-FABP mRNA水平显著升高,而PTEN mRNA表达显著降低(<0.