Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan.
Department of Nutrition, Chung Shan Medical University, Taichung, Taiwan.
Oxid Med Cell Longev. 2018 May 10;2018:2371734. doi: 10.1155/2018/2371734. eCollection 2018.
This study is aimed at developing a method that can inhibit resveratrol (Res) degradation in Dulbecco's modified Eagle medium (DMEM) and at evaluating the effects of Res on the replicative lifespan of Hs68 cells. We hypothesized that Res can extend the lifespan of Hs68 cells if we can inhibit the oxidative degradation of Res in the medium. We found that the addition of ≥5 U/mL SOD to the medium could completely inhibit Res degradation in DMEM. Fetal bovine serum (FBS) contained 29.3 ± 1.1 U/mL of SOD activity. FBS could prevent Res degradation in the medium through SOD activity and Res-FBS interaction, but the regular FBS concentration (i.e., 10% FBS) exhibited an insufficient ability to completely inhibit Res degradation. We found that pyruvate (1 mM) could potentiate SOD to scavenge superoxide at approximately 2.2-fold. Thus, 10% FBS combined with pyruvate (1 mM) could completely inhibit Res degradation. When Res was not degraded, it still shortened the lifespan of Hs68 cells. Overall, the proposed method involving 10% FBS combined with pyruvate (1 mM) could completely prevent Res degradation. However, in contrast to our hypothesis, Res could induce the shortening of the lifespan of Hs68 cells. The stability of Res analogs (i.e., oxy-Res and acetyl-Res) in the medium and their effects on the lifespan of Hs68 cells were also investigated.
本研究旨在开发一种方法,以抑制杜氏改良伊格尔培养基(DMEM)中白藜芦醇(Res)的降解,并评估 Res 对 Hs68 细胞复制寿命的影响。我们假设,如果我们能抑制培养基中 Res 的氧化降解,Res 可以延长 Hs68 细胞的寿命。我们发现,向培养基中添加≥5 U/mL 的 SOD 可完全抑制 DMEM 中 Res 的降解。胎牛血清(FBS)含有 29.3±1.1 U/mL 的 SOD 活性。FBS 可以通过 SOD 活性和 Res-FBS 相互作用来防止培养基中 Res 的降解,但常规 FBS 浓度(即 10% FBS)抑制 Res 降解的能力不足。我们发现,1mM 丙酮酸可以使 SOD 增强约 2.2 倍来清除超氧自由基。因此,10% FBS 与 1mM 丙酮酸结合可完全抑制 Res 的降解。当 Res 不被降解时,它仍然缩短了 Hs68 细胞的寿命。总体而言,所提出的方法涉及 10% FBS 与 1mM 丙酮酸结合,可以完全防止 Res 的降解。然而,与我们的假设相反,Res 可以诱导 Hs68 细胞寿命缩短。还研究了 Res 类似物(即氧基 Res 和乙酰基 Res)在培养基中的稳定性及其对 Hs68 细胞寿命的影响。