Cannabinoid-mediated neuroprotection following interferon-gamma treatment in a three-dimensional mouse brain aggregate cell culture
Cannabinoid-mediated neuroprotection following interferon-gamma treatment in a three-dimensional mouse brain aggregate cell culture
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DOI：
10.1111/j.1460-9568.2004.03711.x
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发表时间：
2004-11-01
影响因子：
3.4
通讯作者：
Diemel, LT
中科院分区：
文献类型：
--
作者：
Jackson, SJ;Baker, D;Diemel, LT
多发性硬化症越来越被认为是一种由中枢神经系统（CNS）炎症引发的神经退行性疾病。脱髓鞘相关的轴突或神经元损伤是残疾的主要原因，并且迄今为止尚未被可用的药物疗法成功靶向。内源性和施用的大麻素的神经保护特性已经在兴奋性毒性、氧化性、创伤性和缺血性损伤后的CNS损伤的体内和体外模型中显示，具有主要的凋亡效应器机制。在这项研究中，胎儿小鼠端脑聚集细胞培养系统的开发，比较大麻素受体1基因敲除小鼠与野生型对应的组织。聚集体形成和神经丝/髓鞘碱性蛋白蓄积取决于胎仔解剖年龄和所用种属。用干扰素-γ治疗后，在体外端脑组织中评估髓鞘碱性蛋白、神经丝、神经元去磷酸化和半胱天冬酶3活化的水平。细胞因子治疗导致大麻素受体1敲除培养物中神经元标记物neuroadherent-H的显著损失，但在野生型中没有，表明大麻素受体1基因的存在可以是神经保护性的。在基因敲除动物的培养物中，胱天蛋白酶3的活化更高，表明细胞死亡的凋亡机制。敲除小鼠中去磷酸化神经丝水平显著升高，支持神经丝去磷酸化是神经元损伤的标志物的前提。总之，这些结果表明，可以通过大麻素受体1引起神经保护，并指出大麻素化合物在脱髓鞘疾病如多发性硬化症中的潜在治疗作用。
Multiple sclerosis is increasingly recognized as a neurodegenerative disease which is triggered by inflammation in the central nervous system (CNS). Demyelination-associated axonal or neuronal damage is a primary cause of disability and has thus far not been successfully targeted by available drug therapies. The neuroprotective properties of both endogenous and administered cannabinoids have been shown in in vivo and in vitro models of CNS damage following excitotoxic, oxidative, traumatic and ischaemic insults, with a predominantly apoptotic effector mechanism. In this study a foetal mouse telencephalon aggregate cell culture system was developed to compare tissue from cannabinoid receptor 1 knockout mice with wildtype counterparts. Aggregate formation and neurofilament/myelin basic protein accumulation were dependent on the age of foetal dissection and species used. Following treatment with interferon-gamma, levels of myelin basic protein, neurofilament, neuronal dephosphorylation and caspase 3 activation were assessed in telencephalon tissue in vitro. Cytokine treatment resulted in significant loss of the neuronal marker neurofilament-H in cannabinoid receptor 1 knockout cultures but not in wildtypes, indicating that presence of the cannabinoid receptor 1 gene can be neuroprotective. Caspase 3 activation was higher in cultures from knockout animals, indicating an apoptotic mechanism of cell death. Dephosphorylated neurofilament levels were significantly elevated in knockout mice, lending support to the premise that neurofilament dephosphorylation is a marker for neuronal damage. Taken together, these results indicate that neuroprotection could be elicited through the cannabinoid receptor 1, and point towards a potential therapeutic role for cannabinoid compounds in demyelinating conditions such as multiple sclerosis.