Fukai Moto, Nakayabu Takuya, Ohtani Shintaro, Shibata Kengo, Shimada Shingo, Sakamoto Soudai, Fuda Hirotoshi, Furukawa Takayuki, Watanabe Mitsugu, Hui Shu-Ping, Chiba Hitoshi, Shimamura Tsuyoshi, Taketomi Akinobu
Department of Gastroenterological Surgery I, Graduate School of Medicine, Hokkaido University, Nishi 7, Kita 15, Kita-ku, Sapporo 060-8638, Hokkaido, Japan.
Faculty of Health Sciences, Graduate School of Health Sciences, Hokkaido University, Nishi5, Kita12, Kita-ku, Sapporo 060-0812, Hokkaido, Japan.
J Clin Med. 2021 May 4;10(9):1972. doi: 10.3390/jcm10091972.
Cold preservation in University of Wisconsin (UW) solution is not enough to maintain the viability of the small intestine, due to the oxidative stress. The novel phenolic antioxidant 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA) has dual properties to reduce oxidative stress, radical scavenging, and antioxidant protein induction, in other cells. This study was designed to determine whether DHMBA reduces cold preservation injury of enterocytes, and to identify the effector site. Enterocytes were subjected to 48-h cold preservation under atmosphere in UW solution (±DHMBA), and then returned to normal culture to replicate reperfusion of the small intestine after cold preservation. At the end of cold preservation (ECP) and at 1, 3, 6, and 72 h after rewarming (R1h, R3h, R6h, and R72h), we evaluated cell function and the injury mechanism. The results showed that DHMBA protected mitochondrial function mainly during cold preservation, and suppressed cell death after rewarming, as shown by the MTT, ATP, mitochondrial membrane potential, LDH, and lipid peroxidation assays, together with enhanced survival signals (PI3K, Akt, p70S6K) and induction of antioxidant proteins (HO-1, NQO-1, TRX-1). We found that DHMBA mitigates the cold-induced injury of enterocytes by protecting the mitochondria through direct and indirect antioxidative activities.
由于氧化应激，在威斯康星大学（UW）溶液中进行冷保存不足以维持小肠的活力。新型酚类抗氧化剂3,5-二羟基-4-甲氧基苄醇（DHMBA）在其他细胞中具有降低氧化应激、清除自由基和诱导抗氧化蛋白的双重特性。本研究旨在确定DHMBA是否能减轻肠细胞的冷保存损伤，并确定其作用位点。将肠细胞在UW溶液（±DHMBA）中于大气环境下进行48小时冷保存，然后恢复正常培养以模拟冷保存后小肠的再灌注。在冷保存结束时（ECP）以及复温后1、3、6和72小时（R1h、R3h、R6h和R72h），我们评估了细胞功能和损伤机制。结果表明，如MTT、ATP、线粒体膜电位、LDH和脂质过氧化测定所示，DHMBA主要在冷保存期间保护线粒体功能，并在复温后抑制细胞死亡，同时增强了存活信号（PI3K、Akt、p70S6K）并诱导了抗氧化蛋白（HO-1、NQO-1、TRX-1）。我们发现，DHMBA通过直接和间接的抗氧化活性保护线粒体，从而减轻肠细胞的冷诱导损伤。
