白肉灵芝水提物对脑缺血大鼠海马神经元的保护作用

doi:10.16431/j.cnki.1671-7236.2021.10.039

摘要/Abstract

摘要： 试验旨在探讨白肉灵芝水提物（Ganoderma leucocontextum aqueous extracts，GLAE）对脑缺血后海马神经元的保护作用及机制。将50只健康大鼠分为对照组、模型组、GLAE低（0.05 mg/（g·BW））、中（0.1 mg/（g·BW））、高（0.2 mg/（g·BW））剂量组。利用双侧颈总动脉夹闭法建立大鼠脑缺血模型，GLAE组灌胃不同剂量的GLAE干预，对照组和模型组灌胃同体积的生理盐水，连续2周。用跳台试验方法检测记忆获得、记忆巩固和记忆再现障碍大鼠的学习记忆能力，HE染色观察大鼠海马组织的病理形态的变化，比色法检测海马组织一氧化氮合酶（nitric oxide synthase，NOS）活性和一氧化氮（nitric oxide，NO）含量，Western blotting和实时荧光定量PCR法分别检测海马组织生长相关蛋白-43（growth associated protein-43，GAP-43）和脑源性神经生长因子（brain derived neurotrophic factor，BDNF）的水平。结果显示，与对照组相比，模型组大鼠跳台试验的逃避潜伏期显著缩短、电击次数显著增加（P<0.05）；海马神经元细胞出现明显核固缩、排列松散紊乱等退行性改变，细胞数量显著减少（P<0.05）；海马组织NOS活性和NO含量均显著降低（P<0.05）；大鼠海马组织GAP-43蛋白表达量显著升高（P<0.05）；海马组织BDNF mRNA表达量显著下调（P<0.05）。与模型组相比，GLAE干预后，大鼠逃避潜伏期均显著延长、电击次数均显著减少（P<0.05）；GLAE高剂量组大鼠CA1区和齿状回锥体神经元细胞形态明显改善，神经元数量显著增加（P<0.05）；GLAE低剂量组对NOS活性影响不明显（P>0.05），显著增加NO含量（P<0.05），GLAE中、高剂量组NOS活性和NO含量均显著升高（P<0.05）；GLAE低、中、高剂量组海马组织GAP-43蛋白表达量均显著增加（P<0.05）；GLAE低、中、高剂量组海马组织BDNF mRNA表达量均显著增加（P<0.05）。以上结果表明，GLAE可通过提高NOS活性和NO水平、促进海马神经发生和功能恢复对脑缺血后海马神经元损伤有一定的保护作用，从而改善大鼠认知功能，0.2 mg/g GLAE效果最好。

关键词: 白肉灵芝水提物; 脑缺血; 大鼠; 一氧化氮合酶(NOS); 生长相关蛋白-43(GAP-43); 脑源性神经生长因子(BDNF)

Abstract: The aim of this study was to investigate the protective effect and mechanism of Ganoderma leucocontextum (GLAE) aqueous extracts on hippocampal neurons after cerebral ischemia. Fifty rats were divided into control, model, low-(0.05 mg/(g·BW)), middle-(0.1 mg/(g·BW)) and high-dose (0.2 mg/(g·BW)) GLAE groups. The cerebral ischemia model of rats was established by bilateral common carotid artery occlusion, and then treated intragastrically with different doses of GLAE once a day for two weeks, and rats of control and model groups were administered the same volume of normal saline. Step down test was used to assess memory acquisition, memory consodilidation and memory reappearance disorder in rats learning and memory ability. The rats pathological changes in hippocampus were observed by HE staining. The nitric oxide synthase(NOS) activity and nitric oxide (NO) content were determined by colorimetric method. The levels of growth associated protein-43 (GAP-43) and brain derived neurotrophic factor (BDNF) in hippocampus were detected by Western blotting and Real-time quantitative PCR, respectively. The results showed that when compared to control group, in the model group, the escape latency of rats in step down test were significantly shortened and the number of electric shocks were significantly increased (P<0.05), hippocampal neurons exhibited obviously degenerative changes with pyknosis and a loose and disorderly arrangement, and a decrease in cell number (P<0.05), NOS activity and NO content were significantly decreased (P<0.05), the level of GAP-43 protein of hippocampal tissue was significantly increased (P<0.05), BDNF mRNA level of hippocampal tissue was significantly decreased (P<0.05). When compared to model group, GLAE administration could obviously prolong the escape latency of rats and decrease significantly the number of electric shocks (P<0.05), the morphology of neurons in GLAE high-dose group of CA1 area and dentate gyrus was significantly improved, and the number of neurons was significantly increased (P<0.05), GLAE low-dose group had no significant effect on NOS activity (P>0.05), but significantly increased the content of NO (P<0.05), while the activity of NOS and the content of NO were significantly increased in middle- and high-dose groups of GLAE (P<0.05). The expression of GAP-43 protein in hippocampus of GLAE low-, middle- and high-dose groups were significantly increased (P<0.05), the level of BDNF mRNA of GLAE low-, middle- and high-dose groups were significantly increased (P<0.05). The above results indicated that GLAE had a certain protective effect on hippocampal neurons injury after cerebral ischemia by increasing the activity of NOS and the content of NO, promoting the hippocampal neurogenesis and neuro-functional recovery, and could improve cognitive function of rats. The effect of 0.2 mg/g GLAE was the best.

Key words: Ganoderma leucocontextum aqueous extracts; cerebral ischemia; rats; nitric oxide synthase (NOS); growth associated protein-43(GAP-43); brain derived neurotrophic factor(BDNF)

中图分类号:

R285

王昱, 秦序, 何九军. 白肉灵芝水提物对脑缺血大鼠海马神经元的保护作用[J]. 中国畜牧兽医, 2021, 48(10): 3864-3871.

WANG Yu, QIN Xu, HE Jiujun. Protective Effect of Ganoderma leucocontextum Aqueous Extracts on Rat Hippocampal Neurons After Cerebral Ischemia[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(10): 3864-3871.

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