国际麻醉学与复苏杂志   2015, Issue (4): 1-1
    
白芦藜醇对小鼠脓毒症相关性脑病炎性损伤的保护作用
隋大鸣, 王嘉锋, 邓小明1()
1.上海市第二军医大学附属长海医院麻醉科
The protective effect of resveratrol on the cerebral inflammatory damage in mice with sepsis-associated encephalopathy
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摘要:

目的 探讨沉默信息调控因子1(sirtuin 1, Sirt1)的天然激动剂白芦藜醇(resveratrol, Res)对脓毒症相关性脑病(sepsis-associated encephalopathy, SAE)炎性损伤的保护作用。 方法 采用盲肠结扎穿孔(cecal ligation and puncture, CLP)方法制作脓毒症小鼠模型,18只C57B/L小鼠按随机数字表法分为3组,每组6只:假手术组(Sham组)、CLP脓毒症模型组(CLP组)以及CLP+Res注射组(Res组)。术后24 h脑组织取材,末端脱氧核糖核苷酸转移酶介导的缺口末端标记法(terminal-deoxynucleotidyl transferase mediated nick end labelling, TUNEL)观察细胞凋亡情况;分离皮质和海马,采用实时定量聚合酶链式反应(real-time polymerase chain reaction, RT-PCR)分别检测Sirt1、含热蛋白结构域NOD样受体家族蛋白-3(NLR family pyrin domain containing-3 protein, NALP3)和白细胞介素(interleukin, IL)-1β的mRNA含量。 结果 TUNEL 染色后电镜下观察,海马Res组阳性细胞数量少于CLP组,凋亡细胞多集中于海马CA1区。Res组小鼠海马和皮质的Sirt1 mRNA的相对表达水平分别为4.62±0.64和4.60±0.61,显著高于Sham组和CLP组(P<0.01)。Res组海马和皮质的NALP3相对表达水平分别为0.86±0.26和0.94±0.35,显著低于CLP组(P<0.05);与Sham组比较,差异无统计学意义 (P>0.05)。Res组海马的IL-1β mRNA相对表达水平为0.57±0.17,显著低于Sham组和CLP组,差异有统计学意义(P<0.01);Res组皮质的IL-1β mRNA相对表达水平为1.61±0.44,与Sham组和CLP组相似,差异无统计学意义(P>0.05)。 结论 SAE时IL-1β在海马区引起的炎性损伤较皮质区明显,Sirt1激动剂白芦藜醇对SAE炎性损伤具有保护作用。

关键词: 白芦藜醇; 脓毒症; 脑病; 盲肠结扎穿孔; 白细胞介素-1β
Abstract:

Objective To investigate the protective effect of sirtuin 1(Sirt 1) agonist, resveratrol(Res), on the cerebral inflammatory damage in mouse with sepsis-associated encephalopathy(SAE). Methods Sepsis model was induced by cecal ligation and puncture(CLP). A total of 18 male C57BL/6 mice were divided into 3 groups randomly by the table of randomization: Sham group, CLP group and Res group(n=6). The brains were harvested 24 h after operation and apoptotic cell death was evaluated by terminal-deoxynucleotidyl transferase mediated nick end labelling(TUNEL) staining. The levels of Sirt1, NLR family pyrin domain containing-3 protein(NALP3) and interleukin(IL)-1β in hippocampus or cortex were measured using real-time polymerase chain reaction(RT-PCR). Results The number of TUNEL positive apoptotic cells in the hippocampal region was significantly less in Res group than that in CLP group. The apoptotic cells were more present in CA1 regions than in the cortex. The RT-PCR results showed that the expression levels of Sirt1 mRNA in hippocampus and cortex in Res group were 4.62±0.64 and 4.60±0.61 respectively, significantly higher than that in Sham group and CLP group(P<0.01). Levels of NALP3 mRNA in hippocampus and cortex in Res group were 0.86±0.26 and 0.94±0.35 respectively, which were significantly lower than that in the CLP group(P<0.05), but similar to that in Sham group. IL-1β mRNA in hippocampus in Res group was 0.57±0.17, significantly lower than in Sham group and CLP group(P<0.01). In cortex, IL-1β in Res group was 1.61±0.44, which was similar to that in Sham group and CLP group(P>0.05). Conclusions Inflammation damage mediated by IL-1β occurs during SAE mainly in the hippocampal region compared with that in cortex and resveratrol plays a protective role against SAE.

Key words: Resveratrol; Sepsis; Encephalopathy; Cecal ligation and puncture; Interleukin-1β