Gok Muslum, Zeybek N Dilara, Bodur Ebru
Hacettepe University, Faculty of Medicine, Department of Medical Biochemistry, 06100, Ankara, Turkey.
Hacettepe University, Faculty of Medicine, Department of Histology and Embryology, 06100, Ankara, Turkey.
Chem Biol Interact. 2016 Nov 25;259(Pt B):276-281. doi: 10.1016/j.cbi.2016.04.029. Epub 2016 Apr 20.
Butyrylcholinesterase (BChE) is mostly associated with the detoxification of xenobiotics. In this study to analyze the involvement of BChE in lipid metabolism, linoleic acid (LA) and α-linolenic acid (ALA) were applied to HepG2 cells along with expression of wild type human BChE. After 48 h of these treatments WST-1 cell proliferation assay, FACS analysis, RT-PCR, Oil Red O staining and activity assays were performed. Application of high concentrations of LA to HepG2 cells without BChE transfection lead to detachment of the cells. The IC value LA was found as 149.3 μM whereas the IC value for ALA could not be calculated. Hence, in order to display minimal effects on cell viability, 5 μM was chosen as appropriate concentration for LA and ALA application to HepG2 cells. Transfection of wild-type BChE plasmid to HepG2 cells yielded increased BChE expression. Application of 5 μM ALA after BChE transfection to HepG2 cells resulted in increased expression of BChE. Although with this low concentration the number of apoptotic cells was decreased with ALA treatments, LA application did not cause a similar result with the same dose. Moreover ghost cell like property was observed in LA-treated cells. Application of ALA, on the other hand, led to an overall increase in cell numbers, BChE expression and activity. Our results indicate that BChE expression might be regulated by ALA in HepG2 cells.
丁酰胆碱酯酶(BChE)主要与外源性物质的解毒作用相关。在本研究中,为了分析BChE在脂质代谢中的作用,将亚油酸(LA)和α-亚麻酸(ALA)与野生型人BChE的表达一起应用于HepG2细胞。经过48小时的这些处理后,进行了WST-1细胞增殖测定、流式细胞术分析、逆转录聚合酶链反应(RT-PCR)、油红O染色和活性测定。在未转染BChE的情况下,对HepG2细胞应用高浓度的LA会导致细胞脱离。发现LA的半数抑制浓度(IC)值为149.3 μM,而ALA的IC值无法计算。因此,为了对细胞活力显示最小影响,选择5 μM作为LA和ALA应用于HepG2细胞的合适浓度。将野生型BChE质粒转染到HepG2细胞中可使BChE表达增加。在将BChE转染到HepG2细胞后应用5 μM ALA导致BChE表达增加。尽管在这种低浓度下,ALA处理使凋亡细胞数量减少,但相同剂量的LA应用并未产生类似结果。此外,在LA处理的细胞中观察到了类似空泡细胞的特性。另一方面,ALA的应用导致细胞数量、BChE表达和活性总体增加。我们的结果表明,在HepG2细胞中,ALA可能调节BChE的表达。