Evidence for simvastatin anti-inflammatory actions based on quantitative analyses of NETosis and other inflammation/oxidation markers.
Evidence for simvastatin anti-inflammatory actions based on quantitative analyses of NETosis and other inflammation/oxidation markers.
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DOI：
10.1016/j.rinim.2014.03.001
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发表时间：
2014-01-01
期刊：
影响因子：
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通讯作者：
Ali, Ashraf
中科院分区：
文献类型：
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作者：
Al-Ghoul, Walid M;Kim, Margarita S;Ali, Ashraf
辛伐他汀（SMV）除了具有经典的降胆固醇作用外，还被证明具有良好的抗炎特性。我们在一个主要的热损伤小鼠模型（三度烫伤，约20%体表面积）中测试了这些新出现的效应，该模型先前已记录有炎症介导的肠道缺陷。中性粒细胞胞外陷阱（NETs）炎症测量方法与经典的肠道黏膜炎症和通透性测量方法一起使用，外源性褪黑素治疗作为阳性对照。我们的假设是辛伐他汀对烧伤后早期肠道黏膜炎症和通透性具有保护性治疗作用。为了验证这一假设，我们将未治疗的热损伤（TI）成年雄性小鼠与烧伤后立即以及在第二天处死前两小时接受辛伐他汀（0.2mg/kg腹腔注射，TI + SMV）治疗的同窝TI小鼠进行比较；将接受褪黑素治疗（Mel）（1.86mg/kg腹腔注射，TI + Mel）的小鼠作为阳性对照进行比较。对小鼠进行了以下评估：（1）使用经典的羰基、Gr - 1和髓过氧化物酶免疫组化或生化分析，检测末端回肠黏膜中的组织氧化和中性粒细胞浸润情况，（2）利用对替代NETosis生物标志物吖啶橙和Gr - 1的流式细胞术分析，检测末端回肠和结肠黏膜匀浆以及腹腔和血液样本中的NETosis情况，（3）用异硫氰酸荧光素 - 葡聚糖和跨上皮电阻（TEER）测量末端回肠和结肠中的跨上皮肠道通透性。我们的结果显示，辛伐他汀和褪黑素对以下情况表现出持续可比的治疗保护作用：（1）如末端回肠中蛋白质羰基化标记以及髓过氧化物酶（MPO）和Gr - 1浸润所显示的肠道黏膜氧化应激，（2）如利用吖啶橙和Gr - 1流式细胞术及显微镜在肠道环境、腹腔灌洗液和血浆中所显示的NETosis，（3）如通过异硫氰酸荧光素 - 葡聚糖渗漏和TEER在回肠和结肠中所评估的跨上皮肠道通透性。因此，辛伐他汀表现出强烈的急性抗炎作用，与肠道组织和全身性NETosis的显著减少以及肠道黏膜通透性降低相关。
Simvastatin (SMV) has been shown to exhibit promising anti-inflammatory properties alongside its classic cholesterol lowering action. We tested these emerging effects in a major thermal injury mouse model (3rd degree scald, ~20% TBSA) with previously documented, inflammation-mediated intestinal defects. Neutrophil extracellular traps (NETs) inflammation measurement methods were used alongside classic gut mucosa inflammation and leakiness measurements with exogenous melatonin treatment as a positive control. Our hypothesis is that simvastatin has protective therapeutic effects against early postburn gut mucosa inflammation and leakiness. To test this hypothesis, we compared untreated thermal injury (TI) adult male mice with TI littermates treated with simvastatin (0.2mg/kg i.p., TI+SMV) immediately following burn injury and twohours before being sacrificed the day after; melatonin-treated (Mel) (1.86mg/kg i.p., TI +Mel) mice were compared as a positive control. Mice were assessed for the following: (1) tissue oxidation and neutrophil infiltration in terminal ileum mucosa using classic carbonyl, Gr-1, and myeloperoxidase immunohistochemical or biochemical assays, (2) NETosis in terminal ileum and colon mucosa homogenates and peritoneal and fluid blood samples utilizing flow cytometric analyses of the surrogate NETosis biomarkers, picogreen and Gr-1, and (3) transepithelial gut leakiness as measured in terminal ileum and colon with FITC-dextran and transepithelial electrical resistance (TEER). Our results reveal that simvastatin and melatonin exhibit consistently comparable therapeutic protective effects against the following: (1) gut mucosa oxidative stress as revealed in the terminal ileum by markers of protein carbonylation as well as myeloperoxidase (MPO) and Gr-1 infiltration, (2) NETosis as revealed in the gut milieu, peritoneal lavage and plasma utilizing picogreen and Gr-1 flow cytometry and microscopy, and (3) transepithelial gut leakiness as assessed in the ileum and colon by FITC-dextran leakiness and TEER. Thus, simvastatin exhibits strong acute anti-inflammatory actions associated with marked decreases in gut tissue and systemic NETosis and decreased gut mucosa leakiness.