Apelin—13对脑缺血再灌注损伤作用的初步研究(1)
【摘要】 目的：研究Apelin-13对缺血/再灌注小鼠脑损伤的作用，并初步探讨其对自噬性细胞死亡的影响。方法：取健康雄性CD-1小鼠，随机分为假手术(sham)组、生理盐水(Saline)组、Apelin组；Apelin组和Saline组首先建立脑缺血再灌注(MCAO/R)模型，再灌注即刻给予同侧侧脑室注射Apelin-13或等量生理盐水，然后在再灌注48 h后，进行神经功能方面检测，同时利用2，3，5-氯化三苯基四氮唑(TTC)染色评估脑梗死体积；缺血再灌注24 h和48 h后，采用West-blot法检测各组自噬相关蛋白LC3的表达情况。结果：Apelin-13可显著降低小鼠脑缺血再灌注后神经功能缺损症状，减少脑梗死体积，减低自噬相关蛋白LC3的表达及LC3-Ⅱ/LC3-Ⅰ的比值。结论：Apelin-13对小鼠脑缺血再灌注损伤有一定的保护作用，抑制神经细胞的自噬性死亡可能是其作用机制之一。
【关键词】 Apelin-13； 脑缺血再灌注模型(MCAO/R)； 自噬； LC3
The Effect of Apelin-13 on Mice after Middle Cerebral Artery Occlusion-Reperfusion Injury/KUAI Jin-xia，BAO Hai-jun，LI Zhou-ru，et al.//Medical Innovation of China，2016，13(20)：018-021
【Abstract】 Objective：To observe the effect of apelin-13 on the brain injury induced by middle cerebral artery occlusion-reperfusion(MCAO/R)，and to explore the relationship between apelin-13 and autophagy in MCAO/R. Method：A total of 48 mice were randomly divided into sham，saline and apelin-13 groups.Firstly，mice s MCAO/R model was established using the suture’s method.After MCAO/R， Apelin-13 and saline groups were pretreated with an immediate injection of Apelin-13 and saline into the mice brain lateral ventricle.The effect of neurological deficit scores and the cerebral infarct volume was measured with TTC at 48h after MCAO/R.At last，to determine the role of Apelin-13 on the brain injury induced by MCAO/R，the autophagy associated proteins LC3 were also assessed with western-bloting.Result：Apelin-13 can significantly decreased neural dysfunction and cerebral infarct volume of mice after MCAO/R 48 h.Inadditionally，Apelin-13 also reduce the amount of protein LC3 expression and down-regulated protein LC3-Ⅱ/LC3-Ⅰratio，at 24 h and 48 h after MCAO/R.Conclusion：Apelin-13 preconditioning has obvious protective effect on mice after MCAO/R，and the mechanism may possibly by suppressing neuron autophagy.
【Key words】 Apelin-13； MCAO/R； Autophagy； LC3
First-author’s address：Xuzhou Medical University，Xuzhou 221002，China
doi：10.3969/j.issn.1674-4985.2016.20.005
脑缺血缺氧可引起的神经细胞结构破坏和脑功能的缺损，脑缺血再灌注损伤是脑缺血缺氧后恢复缺血区血供的又一次损伤，也是一种继发性神经元损伤，与神经细胞的继发性神经元死亡密切相关[1]。近年来有研究表明自噬与脑缺血损伤的发生、发展有着密切的关系[2-3]。研究发现脂肪因子Apelin-13能够对神经细胞的损伤发挥保护作用，但其具体作用机制还不甚明了[4-6]。本实验拟通过观察Aplin-13在脑缺血再灌注损伤中是否起到了保护性作用，并通过检测自噬相关蛋白LC3的表达，研究其对神经细胞自噬的影响，探讨其在缺血再灌注脑损伤中的作用机制。
1 材料与方法
1.1 实验动物及分组 共48只健康雄性CD-1小鼠，体重30～35 g(由徐州医学院动物实验中心提供)，随机分为如下各组，实验1：sham组6只、Saline组6只、Apelin-13组6只，共18只用于行为学检测和TTC实验；实验2组：sham组、再灌注24 h Saline组、再灌注48 h Saline组、再灌注24 h Apelin-13组、再灌注48 h Apelin-13组，每组6只，共30只用于Western-Blot测定。, 百拇医药(蒯锦霞 包海军 李周儒 董国凯 殷文江 孙晓明 李姗姗 蔡红星)
【关键词】 Apelin-13； 脑缺血再灌注模型(MCAO/R)； 自噬； LC3
The Effect of Apelin-13 on Mice after Middle Cerebral Artery Occlusion-Reperfusion Injury/KUAI Jin-xia，BAO Hai-jun，LI Zhou-ru，et al.//Medical Innovation of China，2016，13(20)：018-021
【Abstract】 Objective：To observe the effect of apelin-13 on the brain injury induced by middle cerebral artery occlusion-reperfusion(MCAO/R)，and to explore the relationship between apelin-13 and autophagy in MCAO/R. Method：A total of 48 mice were randomly divided into sham，saline and apelin-13 groups.Firstly，mice s MCAO/R model was established using the suture’s method.After MCAO/R， Apelin-13 and saline groups were pretreated with an immediate injection of Apelin-13 and saline into the mice brain lateral ventricle.The effect of neurological deficit scores and the cerebral infarct volume was measured with TTC at 48h after MCAO/R.At last，to determine the role of Apelin-13 on the brain injury induced by MCAO/R，the autophagy associated proteins LC3 were also assessed with western-bloting.Result：Apelin-13 can significantly decreased neural dysfunction and cerebral infarct volume of mice after MCAO/R 48 h.Inadditionally，Apelin-13 also reduce the amount of protein LC3 expression and down-regulated protein LC3-Ⅱ/LC3-Ⅰratio，at 24 h and 48 h after MCAO/R.Conclusion：Apelin-13 preconditioning has obvious protective effect on mice after MCAO/R，and the mechanism may possibly by suppressing neuron autophagy.
【Key words】 Apelin-13； MCAO/R； Autophagy； LC3
First-author’s address：Xuzhou Medical University，Xuzhou 221002，China
doi：10.3969/j.issn.1674-4985.2016.20.005
脑缺血缺氧可引起的神经细胞结构破坏和脑功能的缺损，脑缺血再灌注损伤是脑缺血缺氧后恢复缺血区血供的又一次损伤，也是一种继发性神经元损伤，与神经细胞的继发性神经元死亡密切相关[1]。近年来有研究表明自噬与脑缺血损伤的发生、发展有着密切的关系[2-3]。研究发现脂肪因子Apelin-13能够对神经细胞的损伤发挥保护作用，但其具体作用机制还不甚明了[4-6]。本实验拟通过观察Aplin-13在脑缺血再灌注损伤中是否起到了保护性作用，并通过检测自噬相关蛋白LC3的表达，研究其对神经细胞自噬的影响，探讨其在缺血再灌注脑损伤中的作用机制。
1 材料与方法
1.1 实验动物及分组 共48只健康雄性CD-1小鼠，体重30～35 g(由徐州医学院动物实验中心提供)，随机分为如下各组，实验1：sham组6只、Saline组6只、Apelin-13组6只，共18只用于行为学检测和TTC实验；实验2组：sham组、再灌注24 h Saline组、再灌注48 h Saline组、再灌注24 h Apelin-13组、再灌注48 h Apelin-13组，每组6只，共30只用于Western-Blot测定。, 百拇医药(蒯锦霞 包海军 李周儒 董国凯 殷文江 孙晓明 李姗姗 蔡红星)