Antishock effect of anisodamine involves a novel pathway for activating α7 nicotinic acetylcholine receptor
Antishock effect of anisodamine involves a novel pathway for activating α7 nicotinic acetylcholine receptor
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DOI：
10.1097/ccm.0b013e31819598f5
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发表时间：
2009-02-01
影响因子：
8.8
通讯作者：
Su, Ding-Feng
中科院分区：
文献类型：
--
作者：
Liu, Chong;Shen, Fu-Ming;Su, Ding-Feng
目的：迷走神经刺激通过烟碱乙酰胆碱受体（α 7 nAChR）信号传导抑制促炎细胞因子的产生。山莨菪碱是一种毒蕈碱型乙酰胆碱受体拮抗剂，在我国临床上已用于治疗各种休克，但其作用机制尚不清楚。在这里，我们测试了山莨菪碱是否通过激活α 7 nAChR达到抗休克作用的假设。设计。随机和对照的体外和体内研究。设置。研究实验室和动物设施室。Sprague-Dawley大鼠、昆明小鼠、α 7 nAChR缺陷小鼠和RAW 264.7细胞。用脂多糖（LPS）（15 mg/kg，静脉内）注射Sprague-Dawley大鼠以诱导败血性休克。在山莨菪碱（10 mg/kg，静脉内）给药前10分钟给予选择性α 7 nAChR拮抗剂甲基甘草次酸（10 mg/kg，腹腔内）。在LPS发作后2小时监测平均动脉压并分析细胞因子。在迷走神经切断小鼠和α 7 nAChR缺陷小鼠中，分别评价了山莨菪碱的抗休克作用。用异硫氰酸荧光素标记的α-银环蛇毒素对RAW264.7细胞进行染色，观察荧光强度。小鼠腹腔巨噬细胞经LPS预处理和刺激后，用酶联免疫吸附测定法测定上清液中肿瘤坏死因子（TNF）-α的含量。甲基乌头碱显著拮抗山莨菪碱对LPS诱导的平均动脉压和TNF-α、IL-1 β表达的有益作用。在迷走神经切断小鼠和α 7 nAChR缺陷小鼠中，山莨菪碱的抗休克作用明显减弱。在体外实验中，山莨菪碱显著增强乙酰胆碱对异硫氰酸荧光素标记的α-银环蛇毒素荧光强度和LPS刺激的TNF-α产生的影响。结论：山莨菪碱的抗休克作用与α 7 nAChR依赖的抗炎通路密切相关。(Crit Care Med 2009; 37：634-641）
Objective: Vagus nerve stimulation inhibits proinflammatory cytokine production by signaling through the 0 nicotinic acetylcholine receptor (alpha 7nAChR). Anisodamine, a muscarinic acetylcholine receptor antagonist, has been used clinically in China for treatment of various shocks, but the mechanism was poorly understood. Here, we tested the hypothesis whether anisodamine attained its antishock effect through activation of alpha 7nAChR.Design. Randomized and controlled in vitro and in vivo study.Settings. Research laboratory and animal facility rooms.Subjects. Sprague-Dawley rats, Kunming mice, alpha 7nAChR-deficient mice, and RAW264.7 cells.Interventions. Sprague-Dawley rats were injected with lipopolysaccharide (LPS) (15 mg/kg, intravenous) to induce septic shock. Methyllycaconitine, a selective alpha 7nAChR antagonist, was administered (10 mg/kg, intraperitoneal) 10 minutes before anisodamine (10 mg/kg, intravenous). Mean arterial pressure was monitored and cytokines were analyzed 2 hours after the onset of LPS. In vagotomized mice and alpha 7nAChR-deficient mice, the antishock effect of anisodamine was appraised, respectively. RAW264.7 cells were stained by fluorescein isothiocyanatelabeled-alpha-bungarotoxin and the fluorescence intensity was observed. Mice peritoneal macrophages were pretreated and stimulated with LPS, and tumor necrosis factor (TNF)-alpha in the supernatant was measured by enzyme-linked immunosorbent assay.Measurements and Main Results. Methyllycaconitine significantly antagonized the beneficial effect of anisodamine on mean arterial pressure and TNF-alpha, interleukin-1 beta expression in response to LPS. The antishock effects of anisodamine were markedly attenuated in vagotomized mice and alpha 7nAChR-deficient mice. In vitro, anisodamine significantly augmented the effect of acetylcholine on fluorescence intensity stained with fluorescein isothiocyanate-labeled-alpha-bungarotoxin and TNF-alpha production stimulated with LPS.Conclusion: These findings demonstrate that the antishock effect of anisodamine is intimately linked to alpha 7nAChR-dependent anti-inflammatory pathway. (Crit Care Med 2009; 37:634-641)