A combination of neostigmine and anisodamine protects against ischemic stroke by activating α7nAChR
A combination of neostigmine and anisodamine protects against ischemic stroke by activating α7nAChR
复制标题
新斯的明和山莨菪明的组合通过激活 α7nAChR 预防缺血性中风
DOI：
10.1111/ijs.12458
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发表时间：
2015-07-01
影响因子：
6.7
通讯作者：
Liu, Ai-Jun
中科院分区：
文献类型：
--
作者：
Qian, Jiao;Zhang, Jing-Ming;Liu, Ai-Jun
背景：新斯的明增加内源性乙酰胆碱可降低缺血性脑损伤。对毒蕈碱受体的脱靶作用产生多种不良反应，限制了其在脑卒中中的临床应用。目的：研究新斯的明与山莨菪碱联合用药的神经保护作用及其机制。方法采用Sprague-Dawley大鼠大脑中动脉闭塞法。研究了新斯的明与山莨菪碱不同比例联用的神经保护作用，以确定最佳的联用组合和理想的治疗窗口期。通过测量7个烟碱受体敲除小鼠的梗死面积、促炎细胞因子的表达和细胞凋亡的生物标志物，研究7个烟碱乙酰胆碱受体的潜在参与。我们在RAW264.7细胞中进行了一系列体外实验，探讨可能的分子机制。结果新斯的明/山莨菪碱联合用药具有神经保护作用。这种保护在1:500的比例下最有效。在此比例下，结合增加了乙酰胆碱与7烟碱乙酰胆碱受体的结合，降低了促炎细胞因子。这种神经保护作用仅在野生型小鼠中明显，在7只烟碱乙酰胆碱受体敲除小鼠中不明显。在7种烟碱乙酰胆碱受体野生型小鼠中，联合治疗显著降低了Bad和Bax的表达，升高了Bcl-2和Bcl-xl的表达，而在敲除小鼠中无明显作用。该组合不影响caspase-8、cleaved caspase-8或caspase-12。结论本研究确定了新斯的明与山莨菪碱联合应用对缺血性脑卒中的最佳治疗方案，并提示乙酰胆碱-7烟碱受体参与了缺血性脑卒中的保护作用。
BackgroundIncreasing endogenous acetylcholine by neostigmine decreased the ischemic cerebral injury. The off-target action on muscarinic receptor produced a variety of adverse effects and limited the clinical application on stroke.AimWe combined neostigmine with anisodamine and investigated the neuroprotection and mechanism.MethodsMale Sprague-Dawley rats were subjected to middle cerebral artery occlusion. Neuroprotective action of neostigmine in combination with anisodamine at varying ratios was examined to determine the optimal combination as well as ideal therapeutic window. Potential involvement of 7 nicotinic acetylcholine receptor was examined by measuring the infarct size, the expression of proinflammatory cytokines, and the biomarkers of apoptosis in 7 nicotinic acetylcholine receptor knockout mice. A set of in vitro experiments was conducted in RAW264.7 cells to probe into potential molecular mechanisms.ResultsThe neostigmine/anisodamine combination conferred neuroprotection. The protection was most potent at a ratio of 1:500. At such a ratio, the combination increased the binding of acetylcholine to 7 nicotinic acetylcholine receptor and reduced proinflammatory cytokines. The neuroprotection was evident only in wild-type and not in 7 nicotinic acetylcholine receptor knockout mice. The combination significantly decreased the expression of Bad and Bax, and increased Bcl-2 and Bcl-xl in 7 nicotinic acetylcholine receptor wild-type mice but not in knockout mice. The combination did not affect caspase-8, cleaved caspase-8, or caspase-12.ConclusionsCurrent study identified the optimal combination of neostigmine and anisodamine against ischemic stroke, and indicated that the acetylcholine-7 nicotinic acetylcholine receptor is involved in the protective effects.