校欢;马晓娇;承欧梅;邱红梅;蒋青松;
XIAO Huan;MA Xiao-jiao;CHENG Ou-mei;QIU Hong-mei;JIANG Qing-song;Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology,Department of Pharmacology,Chongqing Medical University;Department of Neurology,the First Affiliated Hospital of Chongqing Medical University;

• 1:重庆医科大学药理学教研室重庆市生物化学与分子药理学重点实验室
• 2:重庆医科大学附属第一医院神经内科

摘要(Abstract)：

研究天麻素(gastrodin,GAS)对脑缺血后海马神经发生的作用,并探讨其与NO相关可能机制。利用双侧颈总动脉夹闭法建立C57BL/6小鼠脑缺血模型,分别用HE染色和Morris水迷宫检测海马CA1区的病理形态及小鼠认知功能;免疫荧光染色检测Brd U/Neu N阳性细胞数;比色法、硝酸还原法分别检测NOS活力和NO含量; ELISA和Western blot法分别检测c GMP水平和PKG蛋白表达。术后8 d,小鼠海马CA1区锥体神经元结构不规则,出现明显核固缩,细胞排列松散,神经元数目明显减少; 29 d空间学习和记忆能力下降。结果提示,小鼠出现了脑缺血损伤。同时,模型小鼠海马齿状回Brd U/Neu N阳性细胞数明显增加,提示脑缺血后海马出现神经发生。不同剂量GAS(50,100 mg·kg-1)处理明显减轻小鼠CA1区的病理形态损伤,改善学习记忆能力,并促进海马神经发生。同时,模型组NOS-NO减少,但c GMP-PKG增加。GAS给药激活NOS,促进NO生成,增加c GMP水平,上调PKG表达。上述结果提示,GAS可促进脑缺血后海马神经发生,改善小鼠认知功能,该作用可能与NO-cGMP-PKG通路的激活有关。
This paper was aimed to investigate the effect of gastrodin( GAS) on hippocampal neurogenesis after cerebral was chemic and to explore its mechanism of action related to NO. The cerebral ischemia model of C57 BL/6 mice was established by bilateral common carotid artery occlusion. The pathological changes in hippocampal CA1 region and the cognitive function of mice were assessed by HE staining and Morris water maze test,respectively. The count of Brd U/Neu N positive cells in dentate gyrus was detected by immunofluorescence assay. The NOS activity and the NO content were determined by colorimetric and nitrate reduction methods,respectively.The level of c GMP was measured by ELISA kit,and the PKG protein expression was tested by Western blot. On postoperative day 8,the hippocampal CA1 pyramidal neurons of mice showed irregular structure,with obvious nuclear pyknosis,loose cell arrangement and obvious decrease in the number of neurons. On postoperative day 29,the spatial learning ability and memory were decreased. These results indicated cerebral ischemia in mice. Meanwhile,the Brd U/Neu N positive cells were increased significantly in ischemic mice,indicating that neurogenesis occurred in hippocampus after cerebral ischemia. Treatment with different doses of gastrodin( 50 and 100 mg·kg-1) significantly ameliorated the pathological damages in the CA1 region,improved the ability of learning and memory,and promoted hippocampal neurogenesis. At the same time,both the NOS activity and the NO concentration were decreased in model group,but the c GMP level was increased,and the PKG protein expression was up-regulated. Gastrodin administration activated the NOS activity,promoted NO production,further increased c GMP level and up-regulated PKG protein expression. These results suggested that gastrodin can promote hippocampal neurogenesis after cerebral ischemia and improve cognitive function in mice,which may be related to the activation of NO-cGMP-PKG signaling pathway.

关键词(KeyWords)： 天麻素;脑缺血;海马;神经发生;NOS;NO;cGMP;PKG
gastrodin;cerebral ischemia;hippocampus;neurogenesis;NOS;NO;cGMP;PKG

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金面上项目(81871002)

作者(Author): 校欢;马晓娇;承欧梅;邱红梅;蒋青松;
XIAO Huan;MA Xiao-jiao;CHENG Ou-mei;QIU Hong-mei;JIANG Qing-song;Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology,Department of Pharmacology,Chongqing Medical University;Department of Neurology,the First Affiliated Hospital of Chongqing Medical University;

Email:

DOI:

参考文献(References)：

• [1] Toda T,Parylak S L,Linker S B,et al. The role of adult hippocampal neurogenesis in brain health and disease[J]. Mol Psychiatry,2019,24(1):67.
• [2] Miles D K,Kernie S G. Hypoxic-ischemic brain injury activates early hippocampal stem/progenitor cells to replace vulnerable neuroblasts[J]. Hippocampus,2008,18(8):793.
• [3] Liang A C,Mandeville E T,Maki T,et al. Effects of aging on neural stem/progenitor cells and oligodendrocyte precursor cells after focal cerebral ischemia in spontaneously hypertensive rats[J]. Cell Transplant,2016,25(4):705.
• [4]杨飞，王信，马传江，等．天麻加工炮制、成分分析与体内代谢研究进展[J]．中国中药杂志，2018,43(11):2207.
• [5]李爽，夏豪，刘浙波，等．天麻素在心脑血管疾病中的研究进展[J]．现代中药研究与实践，2019,33(1):76.
• [6]文欢，张大燕，王伟，等．天麻素对糖氧剥夺再复供皮层神经细胞NF-κB炎症级联信号通路表达的影响[J]．中国实验方剂学杂志，2017,23(21):104.
• [7] Qiu C W,Liu Z Y,Zhang F L,et al. Post-stroke gastrodin treatment ameliorates ischemic injury and increases neurogenesis and restores the Wnt/β-catenin signaling in focal cerebral ischemia in mice[J]. Brain Res,2019,17(12):7.
• [8] Tulsulkar J,Shah Z A. Ginkgo biloba prevents transient global ischemia-induced delayed hippocampal neuronal death through antioxidant and anti-inflammatory mechanism[J]. Neurochem Int,2013,62(2):189.
• [9] Tian L,Nie H,Zhang Y,et al. Recombinant human thioredoxin-1 promotes neurogenesis and facilitates cognitive recovery following cerebral ischemia in mice[J]. Neuropharmacology,2014,77:453.
• [10]王娜梅，赵玉红，解旭东．缺血性脑血管病的治疗现状及进展研究[J]．医学综述，2017,23(19):3822.
• [11] Stradecki-Cohan H M,Cohan C H,Raval A P,et al. Cognitive deficits after cerebral ischemia and underlying dysfunctional plasticity:potential targets for recovery of cognition[J]. J Alzheimers Dis,2017,60(s1):S87.
• [12] Merson T D,Bourne J A. Endogenous neurogenesis following ischaemic brain injury:insights for therapeutic strategies[J]. Int J Biochem Cell Biol,2014,56:4.
• [13] Sun F L,Wang W,Zuo W,et al. Promoting neurogenesis via Wnt/beta-catenin signaling pathway accounts for the neurorestorative effects of morroniside against cerebral ischemia injury[J].Eur J Pharmacol,2014,73(8):214.
• [14]何珊珊，李锐，吴迪，等．天麻素对脑缺血再灌注小鼠海马新生神经元的保护作用[J]．现代生物医学进展，2015,15(32):6241.
• [15] Collado-Alsina A,Ramírez-Franco J,Sánchez-Prieto J,et al.The regulation of synaptic vesicle recycling by c GMP-dependent protein kinase typeⅡin cerebellar granule cells under strong and sustained stimulation[J]. J Neurosci,2014,34(26):8788.
• [16] Santos A I,Carreira B P,Izquierdo-lvarez A,et al. S-nitrosylation of ras mediates nitric oxide-dependent post-injury neurogenesis in a seizure model[J]. Antioxid Redox Signal,2018,28(1):15.
• [17] Podda M V,Piacentini R,Barbati S A,et al. Role of cyclic nucleotide-gated channels in the modulation of mouse hippocampal neurogenesis[J]. PLoS ONE,2013,8(8):e73246.
• [18] lmestig J N E,Marlet I R,Hainsworth A H,et al. Phosphodiesterase 5 inhibition as a therapeutic target for ischemic stroke:a systematic review of preclinical studies[J]. Cell Signal,2017,3(8):39.
• [19] Engels J,Elting N,Braun L,et al. Sildenafil enhances quantity of immature neurons and promotes functional recovery in the developing ischemic mouse brain[J]. Dev Neurosci,2017,39(1/4):287.