Effects of hypothermia on ex vivo microglial production of pro- and anti-inflammatory cytokines and nitric oxide in hypoxic-ischemic brain-injured mice.
Effects of hypothermia on ex vivo microglial production of pro- and anti-inflammatory cytokines and nitric oxide in hypoxic-ischemic brain-injured mice.
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DOI：
10.5114/fn.2014.43786
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发表时间：
2014-06
影响因子：
2
通讯作者：
T. Matsui;H. Kida;Takuya Iha;Tabito Obara;S. Nomura;T. Fujimiya;Michiyasu Suzuki
中科院分区：
文献类型：
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作者：
T. Matsui;H. Kida;Takuya Iha;Tabito Obara;S. Nomura;T. Fujimiya;Michiyasu Suzuki
激活的小胶质细胞产生神经毒性因子，包括促炎细胞因子和一氧化氮（NO），以响应神经元破坏。通过包括低温在内的各种方法抑制小胶质细胞释放这些因子的治疗性抑制被认为在严重脑损伤后具有神经保护作用。我们通过刺激toll样受体（TLRs），研究了低温培养对缺氧缺血性（HI）脑损伤小鼠体外小胶质细胞产生促炎性和抗炎细胞因子和NO的影响，toll样受体（TLRs）在无菌中枢神经系统损伤的病理过程中起重要作用。材料与方法2日龄小鼠结扎右侧颈总动脉，6%氧溶30 min， 37℃放置24 h，分离小胶质细胞，分别用TLR2和TLR4激动剂在33℃和37℃培养。测定培养上清液中细胞因子和NO水平。结果与37°C相比，低温（33°C）降低了6 h肿瘤坏死因子-α （TNF-α：一种促炎细胞因子）和白细胞介素-10 （IL-10）的产生。结论在tlr激活的HI脑损伤小鼠的小胶质细胞中，低温可暂时降低TNF-α、IL-10和NO的产生，这一临床相关发现提示，治疗性低温的神经保护作用与小胶质细胞释放的早期和晚期炎症因子以及晚期抗炎因子的衰减有关。
INTRODUCTION Activated microglia produce neurotoxic factors, including pro-inflammatory cytokines and nitric oxide (NO), in response to neuronal destruction. Therapeutic suppression of microglial release of these factors by various approaches including hypothermia is considered to be neuroprotective after severe brain damage. We examined the effects of hypothermic culture on the production of pro- and anti-inflammatory cytokines and NO in ex vivo microglia that were derived from mice with hypoxic-ischemic (HI) brain injury, through the stimulation of toll-like receptors (TLRs) that play significant roles in the pathological processes underlying a sterile central nervous system injury. MATERIAL AND METHODS Two-day-old mice underwent the right common carotid artery ligation followed by 6% oxygen for 30 min, and thereafter were placed at 37°C for 24 h, after which microglia were isolated and then cultured with TLR2 and TLR4 agonists at 33°C and 37°C. Cytokine and NO levels in culture supernatants were measured. RESULTS Compared with 37°C, hypothermia (33°C) reduced the production of tumour necrosis factor-alpha (TNF-α: a pro-inflammatory cytokine) at 6 h and interleukin-10 (IL-10: an anti-inflammatory cytokine) and NO at 48 h. CONCLUSIONS In TLR-activated microglia that were derived from mice with HI brain injury, hypothermia reduced the production of TNF-α, IL-10, and NO temporally, a clinically relevant finding suggesting that neuroprotection conferred by therapeutic hypothermia is related to attenuation of early-phase and late-phase inflammatory factors as well as that of late-phase anti-inflammatory factor(s) released from microglia.