白肉灵芝水提物改善衰老大鼠认知功能的作用

doi:10.16098/j.issn.0529-1356.2022.06.004

摘要/Abstract

摘要：

目的探讨白肉灵芝水提物（GLAE）改善衰老大鼠认知功能的作用及机制。方法将50只健康大鼠分成对照组、模型组、GLAE低剂量组、GLAE中剂量组和GLAE高剂量组，用D-半乳糖构建衰老SD大鼠模型，同时分别给予不同剂量的（0、50、100 和200 mg/kg）GLAE处理，采用新物体认知和跳台实验方法检测大鼠认知功能的改变，甲苯胺蓝、吉姆萨和HE染色进行病理学观察，彗星电泳检测大脑细胞DNA的损伤，ELISA、Western blotting和Real-time PCR法检测大脑组织蛋白激酶A（PKA）/环磷酸腺苷（cAMP）反应元件结合蛋白（CREB）信号通路相关因子表达。结果与模型组比较，GLAE干预后，大鼠认知功能衰退及大脑皮层病理变化有显著改善，大脑细胞DNA损伤程度显著降低（P<0.05），大脑组织cAMP、脑源性神经生长因子（BDNF）和神经生长因子（NGF）含量显著增高（P<0.01，P<0.05），PKA、BDNF、NGF和CREB mRNA及PKA、BDNF、NGF和CREB蛋白的表达水平显著上升（P<0.01，P<0.05）。结论 GLAE可改善衰老大鼠的认知功能，其机制可能与其激活大脑PKA/CREB信号通路，增加神经营养因子含量，抑制大脑细胞DNA损伤有关。

关键词: 白肉灵芝水提物, D-半乳糖, 认知功能, 蛋白激酶A/环磷酸腺苷反应元件结合蛋白信号通路, 免疫印迹法, 衰老大鼠

Abstract:

Objective To investigate the effects of the aqueous extracts of ganoderma leucocontextum (GLAE) on cognitive decline of aging rats and possible regulation mechanism. Methods Fifty rats were divided into five groups, control group, model group, GLAE low-dose group, GLAE middle-dose group and GLAE high-dose group. Aging SD rat models were made by D-galactose, and then treated continuously with different doses (0, 50, 100, 200 mg/kg) of GLAE. The novel object recognition and step down test were performed to detect the changes of rats cognitive function. The brain tissue was stained with toluidine blue, Giemsa and HE staining and observed. The cerebral cell DNA damage was detected by comet assay. Expressions of protein kinase A (PKA)/cyclic adenosine monophosphate (cAMP)-response element binding protein (CREB) signaling pathway related factors in brain were respectively detected by ELISA, Western blotting and Real-time PCR. Results Compared with the model group, administration of GLAE could obviously alleviate rats cognitive decline and pathological change. The levels of cell DNA damage reduced markedly (P<0.05). The contents of cAMP, brain derived neurotrophic factor (BDNF) and nerve growth factor (NGF), and also expression levels of mRNA and protein of PKA, BDNF, NGF, CREB in the brain increased significantly in each medicated group (P<0.01, P<0.05). Conclusion GLAE can improve cognitive function, and its mechanism may be related to activation of brain PKA/CREB signaling pathway, increase in neurotrophic factor content and inhibition of cell DNA damage.

Key words: Ganoderma leucocontextum aqueous extracts, D-galactose, Cognitive function, Protein kinase A/cyclic adenosine monophosphate-response element binding protein signaling pathway, Western blotting, Aging rat

中图分类号:

R741

王昱秦序何九军王逸璇 . 白肉灵芝水提物改善衰老大鼠认知功能的作用[J]. 解剖学报, 2022, 53(6): 711-718.

WANG Yu QIN Xu HE Jiu-jun WANG Yi-xuan. Effect of the aqueous extracts of ganoderma leucocontextum on cognitive function of aging rats [J]. Acta Anatomica Sinica, 2022, 53(6): 711-718.

参考文献

［1］Li TH, Hu HP, Deng WQ, et al. Ganoderma leucocontextum, a new member of the G. lucidum complex from southwestern China ［J］. Mycoscience, 2015, 56: 81-85.

［2］Xiong Ch, Chen Ch, Chen ZQ, et al. Potentiation of neuritogenic activity of Ganoderma leucocontextum on rat pheochromocytoma cells［J］.Natural Product Research and Development,2016,28(7): 1135-1138(in Chinese)

熊川, 陈诚, 陈祖琴, 等. 白肉灵芝水提物促进 PC-12细胞分化作用研究［J］. 天然产物研究与开发, 2016, 28(7): 1135-1138.

［3］Xiong Ch, Chen Ch, Huang WL, et al. Protective effects of aqueous extracts of ganoderma leucocontextum on H2O2 induced apopotosis in PC12 cells［J］.Journal of Food Safety and Quality, 2016,7(2):662-668.(in Chinese)

熊川, 陈诚, 黄文丽, 等. 白肉灵芝水提物对 H2O2诱导PC-12 细胞凋亡的保护作用［J］. 食品安全质量检测学报, 2016,7(2): 662-668.

［4］Wang Y, He JJ, Zhang ZZh. Effect of the aqueous extracts of ganoderma leucocontextum on aging rats skin［J］. Nat Prod Res Dev, 2019, 31(12): 2131-2136. (in Chinese)

王昱, 何九军, 张宗舟.白肉灵芝水提物对衰老大鼠皮肤的作用研究［J］. 天然产物研究与开发, 2019, 31(12): 2131-2136.

［5］Gao H, Yan P, Zhang S, et al. Long-term dietary alpha-linolenic acid supplement alleviates cognitive impairment correlatewith activating hippocampal CREB signaling in natural aging rats［J］. Mol Neurobiol, 2016, 53(7): 4772-4781.

［6］Sun HL, Wu H, Liu J, et al. Prenatal stress impairs spatial learning and memory associated with lower mRNA level of the CAMKII and CREB in the adult female rat hippocampus［J］. Neurochem Res, 2017, 42 (5): 1496-1503.

［7］Stepanichev M, Onufriev M, Aniol V, et al. Effects of cerebrolysin on nerve growth factor system in the aging rat brain［J］. Restor Neurol Neuros, 2017, 35(6): 571-581.

［8］Molinari C, Morsanuto V, Rugas S, et al. The role of BDNF on aging-modulation markers［J］. Brain Sci, 2020, 10(5): 285-294.

［9］Song J, Pang YY, Gao L, et al. Effects of scutellaria baicalensis georgi flowers on D-galactose induced aging in rats based on serum metabolomics［J］. Acta Pharmaceutica Sinica,2019, 54(3): 533-539. (in Chinese)

宋佳, 庞溢媛, 高丽, 等. 基于血清代谢组学的黄芩花干预D-半乳糖致衰老大鼠作用研究［J］. 药学学报, 2019, 54(3): 533-539.

［10］Paxinos G, Watson C. The Rat Brain in Stereotaxic Coordinates ［M］. 5th ed. San Diego: Academic Press, 2009：50-115.

［11］Wang YCh, Yu ShY, Wang Y, et al. Effects of X-ray on the structure of cerebral temporal lobe cortex and activity of acetylcholinesterase and expression of Bax and epidermal growth factor of filial mice［J］. Acta Anatomica Sinica, 2011, 42(3): 307-313. (in Chinese)

王元春, 俞诗源, 王昱, 等. X线辐射对仔鼠大脑颞叶皮层结构及乙酰胆碱酯酶活性、Bax蛋白和表皮生长因子表达的影响［J］. 解剖学报, 2011, 42(3): 307-313.

［12］Wang Y, Li ChY, Yu ShY, et al. Effects on the structure of cerebral temporal lobe cortex and capability of learning and memory of filial mice after administration of heroin and ephedrine［J］. Acta Anatomica Sinica, 2009, 40(5): 724-731. (in Chinese)

王昱, 李重阳, 俞诗源, 等. 注射海洛因、麻黄素对仔鼠大脑颞叶皮层结构及学习记忆能力的影响［J］. 解剖学杂志, 2009, 40(5): 724-731.

［13］Ding RC, Ji K, Wu DN, et al. Effect of modified ditan decoction on behavior and PKA/CREB signaling pathway in vascular dementia rats［J］. Chinese Journal of Integrated Traditional and Western Medicine, 2020, 21(1): 17-20. (in Chinese)

丁瑞丛, 纪可, 吴东南, 等. 涤痰汤加味方对血管性痴呆大鼠行为学及PKA/CREB 信号通路的影响［J］. 中国中西医结合杂志, 2020, 21(1): 17-20.

［14］Raghunathan R, Hogan JD, Labadorf A, et al. A glycomics and proteomics study of aging and parkinson’s disease in human brain［J］. Scientific Reports, 2020, 10(1): 1277-1340.

［15］Cortes-canteli M, Iadecola C. Alzheimer’s disease and vascular aging ［J］. J Am Coll Cardiol, 2020, 75(8): 942-951.

［16］Emily M, Amber LS. Biological aging and the cellular pathogenesis of huntington’s disease ［J］. J Huntington’s Disease, 2020, 9(6): 1-14.

［17］Shen Y, Gao X, Qin YQ, et al. Flavored sijunzi delays aging of brain tissue of mice via mTOR signaling pathway［J］. Acta Medicinae Universitatis Scientiae et Technologiae Huazhong, 2018, 47(4): 417-420. (in Chinese)

沈莹, 高啸, 覃艳琼, 等. 加味四君子汤通过mTOR信号通路延缓小鼠脑组织衰老的机制研究［J］. 华中科技大学学报(医学版), 2018, 47(4): 417-420.

［18］Grootaert MOJ, Manon M, Lynn R, et al. Vascular smooth muscle cell death, autophagy and senescence in atherosclerosis ［J］. Cardiovasc Res, 2018, 114(4): 622-634.

［19］Li X, Wang L, Zhang SL, et al. Timing-dependent protection of swimming exercise against D-galactose-induced aging-like impairments in spatial learning/memory in rats［J］. Brain Sci, 2019, 9(9): 236-245.

［20］Ma CL, Li L, Yang GM, et al. Neuroprotective effect of gastrodin in methamphetamine-induced apoptosis through regulating cAMP/PKA/CREB pathway in cortical neuron［J］. Hum Exp Toxicol, 2020, 39(8): 1118-1129.

［21］Li X, Guo C, Li Y, et al. Ketamine administered pregnant rats impair learning and memory in offspring via the CREB pathway［J］. Oncotarget, 2017, 8(20): 32433-32449.

［22］Li YM, Zhang WN, Shi RX, et al. Prenatal caffeine damaged learning and memory in rat offspring mediated by ARs/PKA/CREB/BDNF pathway ［J］. Physiol Res, 2018, 67(6): 975-983.

［23］Du Q, Zhu XY, Si J, et al. Angelica polysaccharide ameliorates memory impairment in Alzheimer’s disease rat through activating BDNF/TrkB/CREB pathway［J］. Exp Biol Med, 2020, 245(1): 1-10.

［24］Wang ChY, Guo JY, Guo RJ. Effect of XingPiJieYu decoction on spatial learning and memory and cAMPPKACREBBDNF pathway in rat model of depression through chronic unpredictable stress［J］. BMC Complement Med Ther, 2017, 17(1): 73-82.

[1]	洪晓敏李三强崔钦奕郑润月杨孟利罗仁利李前辉 . 酒精性肝纤维化核因子κB信号通路与性别的差异 [J]. 解剖学报, 2024, 55(1): 55-61.
[2]	魏茜茜李超红赵晨露唐家萍刘玉珍. 大鼠颈上神经节中Ⅰ组代谢型谷氨酸受体表达及慢性间歇性低氧对其的影响 [J]. 解剖学报, 2024, 55(1): 3-9.
[3]	袁蕾杨智伟杨惠刘政海何洁万炜. 三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛 [J]. 解剖学报, 2024, 55(1): 25-31.
[4]	马婷婷陈建红刘爱翠李海宁. 抑制lncRNA TUG1下调核苷酸结合寡聚结构域样受体蛋白1炎症小体在延缓阿尔茨海默病进展的作用 [J]. 解剖学报, 2024, 55(1): 32-42.
[5]	邹成功冯浩陈兵唐辉邵川孙谋杨荣何家全. 创伤性颅脑损伤中神经功能障碍与认知功能损伤的动态变化 [J]. 解剖学报, 2024, 55(1): 43-48.
[6]	吕海燕沈西雅赵富生朱梅. 松茸多糖对 1-甲基-4-苯基-吡啶离子诱导 PC12 细胞损伤的影响 [J]. 解剖学报, 2024, 55(1): 49-54.
[7]	郑丽平刘霞杜晓华吴亚娟刘珊珊 . 脑源性神经营养因子在牦牛与黄牛端脑不同区域的表达与分布 [J]. 解剖学报, 2024, 55(1): 10-16.
[8]	郝永明郭磊周程辉裴汉军李莉赵哲 . 改良腹主动脉缩窄法建立心肌肥厚模型[J]. 解剖学报, 2024, 55(1): 120-124.
[9]	许亚平余悦欣陈金福王柯柯郭志坤 . 高龄大鼠心室肌间质弹性纤维和胶原纤维的结构分布特征及其机制 [J]. 解剖学报, 2023, 54(6): 716-721.
[10]	王文娟丁雨桐苏昊任捷孙艳荣王涵菲陆嘉莉张林倩白钰秦丽华. 低雌激素状态下大鼠海马及纹状体Nogo-A的表达变化[J]. 解剖学报, 2023, 54(6): 620-627.
[11]	张琴杨俊霞蒋青松. 福辛普利对糖尿病视网膜病变小鼠血管紧张素转化酶2和血管内皮生长因子的影响[J]. 解剖学报, 2023, 54(6): 628-634.
[12]	梁盼盼禹爱梅杜静寇文辉王欢欢宋爱霞. 基于c-Jun氨基末端激酶/p38丝裂原活化蛋白激酶信号通路探讨阿魏酸钠对偏头痛大鼠炎症反应的抑制作用[J]. 解剖学报, 2023, 54(6): 652-659.
[13]	徐颖刘斌郑兴何宗钊. 补体C3a受体在盲肠结扎穿孔致脓毒症大鼠认知障碍中的作用及机制 [J]. 解剖学报, 2023, 54(6): 668-675.
[14]	刘哲川李坤马帅男孟家琪王艳芹. 利拉鲁肽对百草枯诱导的小鼠帕金森病模型炎症及线粒体融合/分裂的影响 [J]. 解剖学报, 2023, 54(6): 676-681.
[15]	吴俊波杨杰肖锋李金亮 . 微小RNA-138调控Wnt通路对体外人脑胶质瘤细胞生物学行为的影响 [J]. 解剖学报, 2023, 54(6): 682-688.