| 李振东,郝 翠,凌 勇,李 瑜,李玲玉,王 鹏,王士雷.线粒体大麻素受体1在大鼠海马神经元缺氧复氧损伤中对线粒体分裂的影响[J].,2018,(4):648-651 |
| 线粒体大麻素受体1在大鼠海马神经元缺氧复氧损伤中对线粒体分裂的影响 |
| Effect of Mitochondrial CB1 Receptor on the Mitochondrial Fission in a Rat Hippocampal Neuron Model of Hypoxia/Re-oxygenation Injury |
| 投稿时间：2017-08-23 修订日期：2017-09-19 |
| DOI：10.13241/j.cnki.pmb.2018.04.010 |
| 中文关键词: 线粒体CB1受体 线粒体分裂 海马神经元 缺氧复氧损伤 |
| 英文关键词: Mitochondrial CB1 receptor Mitochondria fission Hippocampal neuron Hypoxia/ re-oxygenation injury |
| 基金项目:国家自然科学基金面上项目(81371448) |
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| 中文摘要: |
| 摘要 目的：探讨线粒体CB1受体(mitochondrial cannabinoid receptor1，mtCB1)在大鼠海马神经元缺氧复氧损伤中对线粒体分裂的影响。方法：原代培养新生的Wistar大鼠海马神经元，将培养至第8天的海马神经元采用随机数字表分为5组(n=60)：正常组(N组)：正常培养，不做任何处理；缺氧复氧组(H/R组)：采用氧糖剥夺法构建海马神经元缺氧复氧损伤模型，缺氧6h，复氧20 h；缺氧复氧组+ACEA+AM251组(H/R+ACEA+AM251组)：缺氧6 h结束后立即加入ACEA和AM251，终浓度分别为1 μmol/L、10 μmol/L，复氧20 h；缺氧复氧+ACEA+ Hemopressin(H/R+ ACEA+ Hemo组)：缺氧6h结束后立即加入ACEA和Hemopressin，终浓度分别为1 μmol/L、10 μmol/L，复氧20 h；缺氧复氧+赋形剂组(H/R+V组)：同样于缺氧6h结束后立即加入二甲基亚砜(DMSO)，终浓度<0.1 %，复氧20 h。使用激光共聚焦显微镜检测细胞内Ca2+的浓度，流式细胞仪检测细胞凋亡率，Western blot检测凋亡诱导因子(AIF)、线粒体分裂相关蛋白Drp1、Fis1，细胞凋亡相关蛋白细胞色素C(Cytc)和Rho相关的卷曲蛋白激酶1(ROCK1)的表达。结果：与N组相比，H/R组、H/R+ACEA+AM251组、H/R+ACEA+Hemo组和H/R+V组的细胞内Ca2+浓度、细胞凋亡率、以及AIF、Drp1、Fis1、Cytc、ROCK1蛋白的表达水平均明显增加(P<0.05)；与H/R组相比，H/R+ ACEA+Hem组上述各检测指标明显降低(P<0.05)，H/R+ACEA+AM251组和H/R+V组各指标比较差异无统计学意义(P>0.05)。结论：线粒体CB1受体(mtCB1受体)可能通过降低细胞内ROS的含量来减少细胞内Ca2+浓度和ROCK1的表达，进而抑制线粒体分裂，并最终减轻海马神经元缺氧复氧损伤。 |
| 英文摘要: |
| ABSTRACT Objective: To evaluate the effect of mitochondrial CB1 receptor (mtCB1) on mitochondrial fission in a rat hippocampal neuron model of hypoxia/re-oxygenation (H/R) injury. Methods: Primarily cultured hippocampal neurons obtained from Wistar rats were divided into 5 groups(n=60) using a random number table: normal group (N group):were cultured in normal medium, without any administration; H/R group: the hippocampal neurons were subjected to oxygen-glucose deprivation (OGD) for 6h followed by re-oxygenation for 20h; H/R+ACEA+AM251 group: ACEA and AM251 were add with respectively final concentration 1 umol/L, 10 umol/L during the 20h re-oxygenation; H/R+ACEA+ Hemopressin group: ACEA and Hemopressin were add into culture medium with respectively final concentration 1 umol/L, 10 umol/L during the re-oxygenation for 20h; H/R + Vehicle group (H/R+V group): DMSO was add into culture medium with final concentration <0.1 % during the 20h re-oxygenation. Laser scanning confocal microscope was used to measure Ca2+ concentration in cytoplasm. The apoptosis rate was tested by flow cytometry. Western blot was adopted to examine the expression of apoptosis-inducing factor (AIF), dynamin-related protein 1(Drp1), fission 1(Fis1), apoptosis-related protein cytochrome c (Cytc), and Rho-associated coiled-coil containing protein kinase (ROCK1). Results: Compared to the N group, the apoptosis rate, Ca2+ concentration, the expression of AIF, Drp1, Fis1, Cytc and ROCK1were significantly increased in the other four groups (P<0.05); Compared to the H/R group, those detection indexes mentioned above were significantly decreased in H/R+ ACEA+ Hemopressin group (P<0.05), and there were no significant differences were observed in H/R+ACEA+AM251 group and H/R+ Vehicle group (P>0.05); Conclusion: The reduction of ROS induced by mtCB1 can alleviate the expression of ROCK1 and Ca2+ concentration in cytoplasm, and inhibit the mitochondrial fission, and eventually attenuate the H/R injury of hippocampal neurons. |
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