Division of Trauma, Critical Care, and Burn at The Ohio State University Wexner Medical Center, The Ohio State University, Columbus, Ohio, USA.
Institute for Diabetes, Obesity and Metabolism and Department of Physiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Surg Infect (Larchmt). 2023 Nov;24(9):788-796. doi: 10.1089/sur.2023.161.
Sirtuin 3 (SIRT3) is a nicotinamide adenine dinucleotide (NAD)-dependent deacetylase that confers resilience to cellular stress by promoting mitochondrial activity. Mitochondrial dysfunction is a major driver of inflammation during sepsis. We hypothesize that expression improves survival in polymicrobial sepsis by mitigating the inflammatory response. knockout (S3KO) and wild-type (WT) mice underwent cecal ligation and puncture (CLP) or sham surgery. mRNA expression was quantified using quantitative polymerase chain reaction (qPCR) and protein expression was quantified using enzyme-linked immunosorbent assay (ELISA). Spectrophotometric assays were used to quantify serum markers of organ dysfunction. For in vitro studies, bone marrow-derived macrophages (BMDMs) were harvested from S3KO and WT mice and treated with lipopolysaccharide (LPS). After CLP, hepatic levels decreased from baseline by nine hours and remained depressed at 24 hours. Peak serum interleukin-6 (IL-6) protein levels were higher in S3KO mice. In LPS-treated BMDMs, mRNA levels peaked earlier in S3KO cells, although peak levels were comparable to WT. Although S3KO mice had decreased median survival after CLP compared with WT, there was no difference in five-day survival or organ dysfunction. Although S3KO mice initially had increased inflammation and mortality, this difference abated with time, and overall survival was comparable between the groups. This pattern is consistent with the timeline of sepsis-induced downregulation in WT mice, and suggests that downregulation occurring in sepsis is at least partially responsible for the initial hyperinflammatory response and subsequent mortality. Our data support upregulation of as a promising therapeutic strategy for further research in sepsis.
Sirtuin 3（SIRT3）是一种烟酰胺腺嘌呤二核苷酸（NAD）依赖性去乙酰化酶，通过促进线粒体活性赋予细胞应激抗性。线粒体功能障碍是脓毒症中炎症的主要驱动因素。我们假设通过减轻炎症反应，表达可改善多微生物脓毒症的存活率。Sirtuin 3 敲除（S3KO）和野生型（WT）小鼠接受盲肠结扎和穿孔（CLP）或假手术。使用定量聚合酶链反应（qPCR）定量测定 mRNA 表达，使用酶联免疫吸附测定（ELISA）定量测定蛋白表达。分光光度法测定用于定量血清器官功能障碍标志物。对于体外研究，从 S3KO 和 WT 小鼠中采集骨髓来源的巨噬细胞（BMDM），并用脂多糖（LPS）处理。在 CLP 后，肝组织中的水平在 9 小时内从基线下降，并且在 24 小时时仍处于下降状态。S3KO 小鼠的血清白细胞介素 6（IL-6）蛋白水平更高。在 LPS 处理的 BMDM 中，S3KO 细胞中的 mRNA 水平更早达到峰值，尽管峰值水平与 WT 相当。尽管与 WT 相比，S3KO 小鼠在 CLP 后中位存活时间缩短，但 5 天存活率或器官功能障碍无差异。尽管 S3KO 小鼠最初具有更高的炎症和死亡率，但这种差异随着时间的推移而减弱，两组之间的总体存活率相当。这种模式与 WT 小鼠中脓毒症诱导的下调的时间一致，这表明脓毒症中发生的下调至少部分是初始过度炎症反应和随后死亡的原因。我们的数据支持上调作为脓毒症进一步研究的有前途的治疗策略。
