脑缺血诱导学习记忆损伤机制及灯盏花素的保护作用
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摘要
随着世界人口日益老龄化，脑血管疾病的发病率逐年升高，并以其居高不下的致死率和致残率给人类健康和生存造成严重威胁。而脑缺血是诱发此类疾病的重要原因之一。因此，探讨脑缺血的损伤机制对预防和治疗缺血性脑血管疾病意义重大。脑缺血发生后引起的病理生理过程包括能量耗竭、血流再通后的再灌注损伤、神经胶质细胞功能的重新安排，离子平衡紊乱，自由基引起的脂质过氧化损伤，炎症反应，以及兴奋性氨基酸等神经递质的改变均可诱导神经细胞死亡，继而导致学习和记忆等脑功能的损伤。但有关脑缺血引起学习记忆的损伤机制目前尚无定论。
本研究通过双侧颈总动脉重复结扎建立小鼠前脑缺血再灌注模型，综合应用水迷宫等行为检测方法、结合石蜡切片技术、琼脂糖凝胶电泳技术、逆转录聚合酶链反应(RT-PCR)RNA扩增等技术，从整体、细胞和分子水平研究了脑缺血引起学习记忆功能损伤的机制，并应用于中药灯盏花素抗脑缺血引起学习记忆功能减退的研究。
本研究首先观察了重复前脑缺血再灌注模型小鼠在新异环境中的自发活动和水迷宫行为测试，发现前脑重复缺血再灌注并没有明显影响小鼠在新异环境中的自发活动和探究行为(P＞0.05)，但可使小鼠的空间学习记忆能力显著下降(P＜0.01)。以银杏提取物(35mg·kg~(-1))为阳性对照，灯盏花素(5，20，80 mg·kg~(-1))治疗后，缺血小鼠学习记忆能力显著增强(P＜0.05，P＜0.01)。
With the aging of the world population growing, the incidence of cerebrovascular diseases is increasing, and the high mortality and morbidity rates of them pose a serious threat to human health and survival. Cerebral ischemia is one of the important reasons for the diseases. Therefore, the investigation of the mechanisms underlying cerebral ischemic damage is very significant for the prevention and treatment of cerebrovascular diseases. The pathological and physiological process caused by cerebral ischemia, including energy exhaust, neurons damage after reperfusion, reorder of glial cells, ion balance disorders, lipid peroxidation caused by free radical damage, inflammation and excitatory amino acid neurotransmitter changes, can induce neuronal cell death, then resulting in the injury of brain functions such as learning and memory. However, the mechanism underlymg cerebral ischemia-induced learning-memory damage has not a verdict at present.
In the present study, Open field behavior, Morris water maze, and paraffin section techniques, agarose gel electrophoresis techniques, reverse transcription polymerase chain reaction (RT-PCR) amplification techniques were used to investigate the mechanism underlying cerebral ischemia-induced learning-memory damage, further to study the protection of breviscapine against repeated cerebral ischemia-reperfusion induced by fastening bilateral common carotid artery.
The spontaneous behavior and learning-memory of repeated ischemia-reperfusion mice were first tested using open field and Morris water maze. The results showed that repeated ischemia-reperfusion did not affect the spontaneous behavior (P>0.05), but impaired learning-memory (P<0.01). Regarding Ginkgo biloba extract (35mg·kg~(-1)) as a positive control, breviscapine (5,20,80mg·kg~(-1)) can dramatically improve the ability of learning-memory after ischemia-reperfusion (P<0.05, P<0.01).
本研究通过双侧颈总动脉重复结扎建立小鼠前脑缺血再灌注模型，综合应用水迷宫等行为检测方法、结合石蜡切片技术、琼脂糖凝胶电泳技术、逆转录聚合酶链反应(RT-PCR)RNA扩增等技术，从整体、细胞和分子水平研究了脑缺血引起学习记忆功能损伤的机制，并应用于中药灯盏花素抗脑缺血引起学习记忆功能减退的研究。
本研究首先观察了重复前脑缺血再灌注模型小鼠在新异环境中的自发活动和水迷宫行为测试，发现前脑重复缺血再灌注并没有明显影响小鼠在新异环境中的自发活动和探究行为(P＞0.05)，但可使小鼠的空间学习记忆能力显著下降(P＜0.01)。以银杏提取物(35mg·kg~(-1))为阳性对照，灯盏花素(5，20，80 mg·kg~(-1))治疗后，缺血小鼠学习记忆能力显著增强(P＜0.05，P＜0.01)。
With the aging of the world population growing, the incidence of cerebrovascular diseases is increasing, and the high mortality and morbidity rates of them pose a serious threat to human health and survival. Cerebral ischemia is one of the important reasons for the diseases. Therefore, the investigation of the mechanisms underlying cerebral ischemic damage is very significant for the prevention and treatment of cerebrovascular diseases. The pathological and physiological process caused by cerebral ischemia, including energy exhaust, neurons damage after reperfusion, reorder of glial cells, ion balance disorders, lipid peroxidation caused by free radical damage, inflammation and excitatory amino acid neurotransmitter changes, can induce neuronal cell death, then resulting in the injury of brain functions such as learning and memory. However, the mechanism underlymg cerebral ischemia-induced learning-memory damage has not a verdict at present.
In the present study, Open field behavior, Morris water maze, and paraffin section techniques, agarose gel electrophoresis techniques, reverse transcription polymerase chain reaction (RT-PCR) amplification techniques were used to investigate the mechanism underlying cerebral ischemia-induced learning-memory damage, further to study the protection of breviscapine against repeated cerebral ischemia-reperfusion induced by fastening bilateral common carotid artery.
The spontaneous behavior and learning-memory of repeated ischemia-reperfusion mice were first tested using open field and Morris water maze. The results showed that repeated ischemia-reperfusion did not affect the spontaneous behavior (P>0.05), but impaired learning-memory (P<0.01). Regarding Ginkgo biloba extract (35mg·kg~(-1)) as a positive control, breviscapine (5,20,80mg·kg~(-1)) can dramatically improve the ability of learning-memory after ischemia-reperfusion (P<0.05, P<0.01).
引文
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