慢性锂拮抗缺血性脑损伤机制的研究
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摘要
脑中风(缺血)是当今社会主要疾病之一，它的发病率高、死亡率高、致残率高；给病人带来极大的痛苦，也给社会带来沉重的负担。积极预防和早期治疗非常重要。大量的实验研究提示缺血后大脑中的神经元再生增加。近期研究发现：原本用作情绪稳定剂的锂盐对缺血性中风有保护作用。抗抑郁的研究发现锂盐可促进神经元再生，但是目前尚不清楚神经再生是否参与了锂盐对缺血性中风的保护作用。本文采用Sprague-Dawley(SD)大鼠的四动脉结扎全脑缺血模型，旨在阐明锂在缺血性脑中风中的作用机制，着重研究锂通过ERK信号通路促进缺血后神经再生和行为损伤的恢复。
1．锂改善短暂性全脑缺血引起的行为学异常
长期慢性锂能通过抑制细胞凋亡在缺血性脑损伤中起保护作用。为了阐明锂能否改善短暂性全脑缺血引起的行为异常，我们研究了锂对大鼠行为学的影响：包括平衡木，高架十字迷宫，开野和Morris水迷宫。我们的结果显示SD大鼠四动脉结扎引起全脑缺血15分钟后第7天，大鼠的存活率降低，一般状态变差，平衡木中的表现变差，高架十字迷宫和开野试验中活动过度，如进入(?)放臂的次数和时间增多，爬格增多，直立和理毛增多等。给与锂之后，没有提高大鼠的存活率，但是能明显改善大鼠的一般状态；而且能抑制缺血引起的过度活跃现象。缺血后第8和第9天，大鼠的学习记忆能力严重受损，在Morris水迷宫中表现为：找到水下平台需要的时间增长，在原有平台象限停留的时间缩短给与锂之后，能明显改善缺血引起的学习记忆能力损伤。组织学分析显示锂能明显降低海马CA1区细胞死亡。我们的研究进一步从整体功能方面证实了锂在脑缺血再灌注损伤中的保护作用。
2锂在短暂性全脑缺血的大鼠中，通过ERK信号通路调节海马中的神经再生，并促进空间学习记忆的恢复
近年的研究证实锂对脑缺血有保护作用；海马中的神经再生是否参与了这种作用还不清楚；ERK信号通路在调节神经再生中起着重要的作用，所以我们研究了脑缺血的大鼠中，锂是否能通过ERK信号通路调节海马中的神经再生，并改善学习记忆。大鼠每天腹腔注射氯化锂(1mmol／kg)，2周后四动脉结扎缺血15分钟。缺血后第6天，或从缺血后第6天到第8天，5-溴2-脱氧尿嘧啶(50mg／kg)每日腹腔注射2次。通过免疫印迹的方法，我们发现锂能促进缺血后ERK1／2的激活；缺血后第7，第21天，与缺血组相比，锂能促进海马齿状回Brdu阳性细胞数量的增加。而且，锂能增加Brdu阳性细胞的存活率。U0126抑制ERK1／2的活性后，锂的作用也被抑制。锂没有明显影响Brdu阳性细胞的神经元和星形胶质细胞分化率。同时，锂能改善Morris水迷宫中大鼠的空间学习记忆能力；而U0126能消除锂对学习记忆的改善。这些结果提示在短暂性全脑缺血的大鼠中，锂能通过ERK信号通路增加海马中细胞的再生和存活，改善行为损伤；提示锂可能联合其他神经保护药物在中风的治疗中有潜在的临床意义。
Ischemic stroke is a leading cause of death and disability worldwide In more than80% of cases, it results from a transient or permanent reduction in cere(?)ral blood flow,caused by occlusion of a cerebral artery by an embolus or loc(?)l thrombosis.Two-thirds of patients survive the initial event but are left with signifi(?)ant degrees ofsensorimotor, cognitive, or other impairment. So it is critical for active (?)revention andearly treatment. Increasing evidence have showed that neurogenesis is i(?)creased in thebrain after ischemia. Furthermore, lithium, a mood stabilizer, car facilitate theneurogenesis and protect the brain against ischemic injury. In the c(?)rent study, toformulate the role of lithium and the underlying mechanisms in ischem c rats inducedby four-vessel occlusion, we examined whether lithium improves he behavioraldisorder in ischemic rats and whether lithium regulates hippocampal n urogenesis byERK pathway. Formulating the role and mechanism of lithium acti(?)ns may havepotential clinical benefits for the treatment of stroke in combination with otherneuroprotective agents.
1. Lithium improves the behavioral disorder in rats subjected to tr(?)nsient globalcerebral ischemia
Previous study has indicated that chronic treatment with lithium protects brainagainst ischemic injury by reducing apoptotic death. To investigate whether lithiumimproves the behavioral disorder induced by transient global cerebra ischemia, weexamined the effects of lithium treatment on the performance of r(?)s in a set ofbehavioral tests, i.e. beam balance, elevated plus maze (EPM), open fi(?)ld and Morriswater maze. Our results showed that lithium attenuated the worse gene(?)l 'well-being'and the worse performance in beam balance, and hyperactivity in EPM and open field,including increased open arm entries, time spent in the open arms, s(?)ares crossed,rearing and grooming over 7 days after 15 min ischemia, which were induced byfour-vessel occlusion in Sprague-Dawley rats. Moreover, lithium improved theinjured spatial learning and memory ability in Morris water maze at post-ischemicdays 8 and 9. Histological analysis displayed that it decreased obvious(?) cell death in##原图像不清晰 hippocampal CA1 region. Our study further confirmed the protective role of lithiumin the ischemia-reperfusion injury and suggested that lithium might be a helpfultherapeutic approach to the treatment of stroke combining with other neuroprotectiveagents.
2. Lithium regulates hippocampal neurogenesis by ERK pathway and facilitatesrecovery of spatial learning and memory in rats after transient global cerebralischemia
Recent studies have demonstrated that lithium has a neuroprotective effect againstbrain ischemia. Whether this effect is mediated by hippocampal neurogenesis remainsunknown. The ERK (extracellular signal-regulated kinase) pathway plays an essentialrole in regulating neurogenesis. The present study was undertaken to investigatewhether lithium regulates hippocampal neurogenesis by the ERK pathway andimproves spatial learning and memory deficits in rats after ischemia. Rats were dailyinjected with lithium (1mmol/kg) and two weeks later subjected to 15-minuteischemia induced by four-vessel occlusion method. 5-bromo-2'-deoxyuridine (Brdu;50mg/kg) was administrated twice daily at postischemic day 6, or for 3 days frompostischemic day 6 to 8. We found that lithium increased the ERK1/2 activation afterischemia by western blotting analysis. There was a significant increase inBrdu-positive cells in the hippocampal dentate gyrus after lithium treatment,compared with ischemia group at postischemic day 7 and 21. And importantly, thesurvival rate of Brdu-positive cells was elevated by lithium. Inhibition of the ERK1/2activation by U0126 diminished these effects of lithium. The percentages ofBrdu-positive cells that expressed a neuronal marker or an astrocytic marker were notsignificantly influenced by lithium. Moreover, lithium improved the impaired spatiallearning and memory ability in Morris water maze, and U0126 attenuated thebehavioral improvement by lithium. These results suggest that lithium up-regulatesthe generation and survival of new-born cells in the hippocampus by the ERKpathway and improves the behavioral disorder in rats after transient global cerebralischemia.
1．锂改善短暂性全脑缺血引起的行为学异常
长期慢性锂能通过抑制细胞凋亡在缺血性脑损伤中起保护作用。为了阐明锂能否改善短暂性全脑缺血引起的行为异常，我们研究了锂对大鼠行为学的影响：包括平衡木，高架十字迷宫，开野和Morris水迷宫。我们的结果显示SD大鼠四动脉结扎引起全脑缺血15分钟后第7天，大鼠的存活率降低，一般状态变差，平衡木中的表现变差，高架十字迷宫和开野试验中活动过度，如进入(?)放臂的次数和时间增多，爬格增多，直立和理毛增多等。给与锂之后，没有提高大鼠的存活率，但是能明显改善大鼠的一般状态；而且能抑制缺血引起的过度活跃现象。缺血后第8和第9天，大鼠的学习记忆能力严重受损，在Morris水迷宫中表现为：找到水下平台需要的时间增长，在原有平台象限停留的时间缩短给与锂之后，能明显改善缺血引起的学习记忆能力损伤。组织学分析显示锂能明显降低海马CA1区细胞死亡。我们的研究进一步从整体功能方面证实了锂在脑缺血再灌注损伤中的保护作用。
2锂在短暂性全脑缺血的大鼠中，通过ERK信号通路调节海马中的神经再生，并促进空间学习记忆的恢复
近年的研究证实锂对脑缺血有保护作用；海马中的神经再生是否参与了这种作用还不清楚；ERK信号通路在调节神经再生中起着重要的作用，所以我们研究了脑缺血的大鼠中，锂是否能通过ERK信号通路调节海马中的神经再生，并改善学习记忆。大鼠每天腹腔注射氯化锂(1mmol／kg)，2周后四动脉结扎缺血15分钟。缺血后第6天，或从缺血后第6天到第8天，5-溴2-脱氧尿嘧啶(50mg／kg)每日腹腔注射2次。通过免疫印迹的方法，我们发现锂能促进缺血后ERK1／2的激活；缺血后第7，第21天，与缺血组相比，锂能促进海马齿状回Brdu阳性细胞数量的增加。而且，锂能增加Brdu阳性细胞的存活率。U0126抑制ERK1／2的活性后，锂的作用也被抑制。锂没有明显影响Brdu阳性细胞的神经元和星形胶质细胞分化率。同时，锂能改善Morris水迷宫中大鼠的空间学习记忆能力；而U0126能消除锂对学习记忆的改善。这些结果提示在短暂性全脑缺血的大鼠中，锂能通过ERK信号通路增加海马中细胞的再生和存活，改善行为损伤；提示锂可能联合其他神经保护药物在中风的治疗中有潜在的临床意义。
Ischemic stroke is a leading cause of death and disability worldwide In more than80% of cases, it results from a transient or permanent reduction in cere(?)ral blood flow,caused by occlusion of a cerebral artery by an embolus or loc(?)l thrombosis.Two-thirds of patients survive the initial event but are left with signifi(?)ant degrees ofsensorimotor, cognitive, or other impairment. So it is critical for active (?)revention andearly treatment. Increasing evidence have showed that neurogenesis is i(?)creased in thebrain after ischemia. Furthermore, lithium, a mood stabilizer, car facilitate theneurogenesis and protect the brain against ischemic injury. In the c(?)rent study, toformulate the role of lithium and the underlying mechanisms in ischem c rats inducedby four-vessel occlusion, we examined whether lithium improves he behavioraldisorder in ischemic rats and whether lithium regulates hippocampal n urogenesis byERK pathway. Formulating the role and mechanism of lithium acti(?)ns may havepotential clinical benefits for the treatment of stroke in combination with otherneuroprotective agents.
1. Lithium improves the behavioral disorder in rats subjected to tr(?)nsient globalcerebral ischemia
Previous study has indicated that chronic treatment with lithium protects brainagainst ischemic injury by reducing apoptotic death. To investigate whether lithiumimproves the behavioral disorder induced by transient global cerebra ischemia, weexamined the effects of lithium treatment on the performance of r(?)s in a set ofbehavioral tests, i.e. beam balance, elevated plus maze (EPM), open fi(?)ld and Morriswater maze. Our results showed that lithium attenuated the worse gene(?)l 'well-being'and the worse performance in beam balance, and hyperactivity in EPM and open field,including increased open arm entries, time spent in the open arms, s(?)ares crossed,rearing and grooming over 7 days after 15 min ischemia, which were induced byfour-vessel occlusion in Sprague-Dawley rats. Moreover, lithium improved theinjured spatial learning and memory ability in Morris water maze at post-ischemicdays 8 and 9. Histological analysis displayed that it decreased obvious(?) cell death in##原图像不清晰 hippocampal CA1 region. Our study further confirmed the protective role of lithiumin the ischemia-reperfusion injury and suggested that lithium might be a helpfultherapeutic approach to the treatment of stroke combining with other neuroprotectiveagents.
2. Lithium regulates hippocampal neurogenesis by ERK pathway and facilitatesrecovery of spatial learning and memory in rats after transient global cerebralischemia
Recent studies have demonstrated that lithium has a neuroprotective effect againstbrain ischemia. Whether this effect is mediated by hippocampal neurogenesis remainsunknown. The ERK (extracellular signal-regulated kinase) pathway plays an essentialrole in regulating neurogenesis. The present study was undertaken to investigatewhether lithium regulates hippocampal neurogenesis by the ERK pathway andimproves spatial learning and memory deficits in rats after ischemia. Rats were dailyinjected with lithium (1mmol/kg) and two weeks later subjected to 15-minuteischemia induced by four-vessel occlusion method. 5-bromo-2'-deoxyuridine (Brdu;50mg/kg) was administrated twice daily at postischemic day 6, or for 3 days frompostischemic day 6 to 8. We found that lithium increased the ERK1/2 activation afterischemia by western blotting analysis. There was a significant increase inBrdu-positive cells in the hippocampal dentate gyrus after lithium treatment,compared with ischemia group at postischemic day 7 and 21. And importantly, thesurvival rate of Brdu-positive cells was elevated by lithium. Inhibition of the ERK1/2activation by U0126 diminished these effects of lithium. The percentages ofBrdu-positive cells that expressed a neuronal marker or an astrocytic marker were notsignificantly influenced by lithium. Moreover, lithium improved the impaired spatiallearning and memory ability in Morris water maze, and U0126 attenuated thebehavioral improvement by lithium. These results suggest that lithium up-regulatesthe generation and survival of new-born cells in the hippocampus by the ERKpathway and improves the behavioral disorder in rats after transient global cerebralischemia.
引文
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