From the FH "Sammy" Ross Trauma Center, Department of Surgery, Carolinas Medical Center, Charlotte, North Carolina.
J Trauma Acute Care Surg. 2014 Feb;76(2):409-17. doi: 10.1097/TA.0000000000000096.
Oxidative stress following hemorrhagic shock and resuscitation (HSR) is regulated, in part, by inflammatory and apoptotic mediators such as necrosis factor κB (NF-κB) and p53. Sirtuin 1 (Sirt-1) is a metabolic intermediary that regulates stress responses by suppressing NF-κB and p53 activity. Resveratrol is a naturally occurring polyphenolic antioxidant and Sirt-1 agonist. The aim of this study was to determine whether resveratrol protects hepatocytes following HSR or hypoxia.
In vivo, HSR was achieved in male rats by arterial blood withdrawal to 30 ± 2 mm Hg for 1 hour before resuscitation with or without resveratrol (Res, 30 mg/kg). Hepatic tissue was stained and scored for necrosis, interleukin 6, and Sirt-1 expression. In vitro, primary rat hepatocytes were subjected to 8 hours of hypoxia without or with Res (100 µM). Cells were analyzed immediately or after 6 hours of normoxia, for survival and markers of injury (lactate dehydrogenase assay, lipid peroxidation, and mitochondrial integrity). Cell lysates were collected for cytochrome c analysis and immunoprecipitated using antibodies against NF-κB (p65) or p53.
In vivo, animals subject to HSR exhibited increased expression of markers of hepatocyte damage compared with those sham operated, concomitant with lower Sirt-1 expression. In vitro, hypoxia followed by normoxia resulted in increased cell death, an effect that was blunted by Res. Analysis of cell and mitochondrial function demonstrated that Res inhibited the detrimental effects of hypoxia in isolated hepatocytes.
Resveratrol prevents cell death in HSR and exerts a protective effect on the mitochondria in a hepatocyte model of hypoxic injury-reoxygenation possibly via Sirt-1 modulation of p53 and NF-κB activity.
出血性休克复苏（HSR）后氧化应激部分受到炎症和凋亡介质的调节，如核因子 κB（NF-κB）和 p53。Sirtuin 1（Sirt-1）是一种代谢中间产物，通过抑制 NF-κB 和 p53 的活性来调节应激反应。白藜芦醇是一种天然存在的多酚抗氧化剂和 Sirt-1 激动剂。本研究旨在确定白藜芦醇是否能在 HSR 或缺氧后保护肝细胞。
在体内，雄性大鼠通过动脉采血至 30±2mmHg 1 小时，然后进行 HSR，复苏时用或不用白藜芦醇（Res，30mg/kg）。对肝组织进行染色并对坏死、白细胞介素 6 和 Sirt-1 表达进行评分。在体外，原代大鼠肝细胞在无或有 Res（100µM）的情况下缺氧 8 小时。细胞立即或在正常氧 6 小时后进行分析，用于检测存活和损伤标志物（乳酸脱氢酶测定、脂质过氧化和线粒体完整性）。收集细胞裂解物进行细胞色素 c 分析，并使用针对 NF-κB（p65）或 p53 的抗体进行免疫沉淀。
体内，与假手术组相比，HSR 动物表现出肝损伤标志物表达增加，同时 Sirt-1 表达降低。体外，缺氧后再氧合导致细胞死亡增加，而 Res 则减弱了这种作用。细胞和线粒体功能分析表明，Res 抑制了缺氧对分离肝细胞的有害影响。
白藜芦醇可预防 HSR 中的细胞死亡，并在缺氧损伤-再氧合的肝细胞模型中对线粒体发挥保护作用，这可能是通过 Sirt-1 调节 p53 和 NF-κB 的活性。
