目的 探讨柑橘橘络提取物对低氧小鼠记忆损伤的保护作用。方法 30只雄性BALB/c小鼠,随机分为常氧对照组（CON）、低氧模型组（HYP）和柑橘橘络提取物组（CIT）,每组10只,将后2组小鼠置于模拟海拔6000 m的低压氧舱内,每天8 h,持续28 d。CON组和HYP组灌胃给予0.4 ml纯净水,CIT组灌胃给予15 mg/(kg·bw)柑橘橘络提取物。采用穿梭和跳台实验评价小鼠的主动学习记忆和被动学习记忆能力;采用苏木精-伊红（hematoxylin-eosin, HE）染色观察海马病理学变化;采用生物化学方法检测脑组织乳酸脱氢酶（lactic dehydrogenase, LDH）和ATP酶活力;采用代谢组学方法靶向检测小鼠脑组织糖酵解和三羧酸循环代谢产物。结果 与CON组相比,HYP组小鼠的主动穿梭次数减少（P<0.05）、无响应次数增加（P<0.05）;跳台实验错误次数增加（P<0.05）、潜伏期缩短（P<0.05）;海马细胞排列松散、核固缩增加;ATP、磷酸烯醇式丙酮酸、乌头酸、异柠檬酸、α-酮戊二酸、琥珀酸和草酰乙酸含量显著降低（P<0.05）,乳酸、6-磷酸葡萄糖、β-6-磷酸果糖和1,6-二磷酸果糖含量显著升高（P<0.05）;LDH和ATP酶活力显著降低（P<0.05）。与HYP组比较,柑橘橘络提取物可以增加穿梭实验主动穿梭次数（P<0.05）,降低无响应次数（P<0.05）;减少跳台实验错误次数（P<0.05）,延长跳下平台的潜伏期（P<0.05）;改善海马细胞细胞形态及排列紧密程度;升高ATP和磷酸烯醇式丙酮酸含量（P<0.05）,降低乳酸、6-磷酸葡萄糖、β-6-磷酸果糖和1,6-二磷酸果糖含量（P<0.05）;降低LDH活力（P<0.05）,增强ATPase活力（P<0.05）。结论 柑橘橘络提取物具有改善低氧小鼠记忆损伤的作用,推测可能与调控脑能量代谢相关。
Abstract
Objective To explore the protective effects of citrus tangerine pith extract on memory impairment in hypoxic mice. Methods Thirty male BALB/c mice were randomised into normoxia control (CON), hypoxia model (HYP) and citrus tangerine pith extract (CIT) groups, with 10 mice in each group. The latter two groups were placed in a low-pressure oxygen chambers at an altitude simulating 6,000 meters for 8 hours per day for 28 days. The CON group and the HPY group were given 0.4 ml of pure water by gavage, and the CIT group was given 15 mg/(kg·bw) of citrus tangerine pith extract. Shuttle experiment and step-down test were used to assess the active and passive learning and memory abilities. Hematoxylin-eosin staining was used to observe the morphological changes in hippocampus tissue. Biochemical methods were used to determine LDH and ATPase activity in brain tissue. The metabolites of the glycolytic and tricarboxylic acid cycles in brain tissue were identified using metabonomics. Results Compared with the CON group, mice in the HYP group showed a looser arrangement of neurons in the hippocampus and an increase in the nuclear consolidation. In the shuttle experiment, the number of active shuttles was significantly decreased (P<0.05), and the number of non-responses was significantly increased (P<0.05). In the step-down test, the number of errors was significantly increased (P<0.05) and the latency was significantly shortened (P<0.05). The contents of ATP, PEP, aconitrate, isocitrate, KGA, SA, and OA were decreased (P<0.05), while the contents of LA, D-G6P, β-D-F6P and F1,6P were increased (P<0.05). The LDH and ATPase activities of brain tissues in the HYP group were significantly lower (P<0.05). Compared with the HYP group, citrus tangerine pith extract significantly elevated the number of active shuttles in the shuttle experiment (P<0.05) and decreased the number of non-responses (P<0.05). In the step-down test, the number of errors was significantly reduced (P<0.05) and the latency was significantly prolonged (P<0.05). Citrus tangerine pith extract improved the cell morphology and tightness of arrangement in the hippocampus. The contents of ATP and PEP were elevated (P<0.05), and the contents of LA, D-G6P, β-D-F6P and F1,6P were reduced (P<0.05). The LDH activity in citrus tangerine pith extract group was lowered (P<0.05), and the ATPase was enhanced (P<0.05). Conclusion Citrus tangerine pith extract can ameliorate memory impairment in hypoxic mice, which is related to the modulation of brain energy metabolism.
关键词
低氧 /
柑橘橘络提取物 /
小鼠 /
记忆损伤 /
能量代谢
Key words
hypoxia /
citrus tangerine pith extract /
mice /
memory impairment /
energy metabolism
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Midha AD, Zhou Y, Queliconi BB, et al. Organ-specific fuel rewiring in acute and chronic hypoxia redistributes glucose and fatty acid metabolism[J]. Cell Metab, 2023, 35: 504–516.
[2] Xu F, Liu P, Pascual JM, et al. Effect of hypoxia and hyperoxia on cerebral blood flow, blood oxygenation, and oxidative metabolism[J]. J Cereb Blood Flow Metab, 2012, 32: 1909–1918.
[3] Li P, Zhang G, You HY, et al. Training-dependent cognitive advantage is suppressed at high altitude[J].Physiol Behav, 2012, 106: 439–445.
[4] Zhou Y, Lu H, Liu Y, et al. Cirbp-PSD95 axis protects against hypobaric hypoxia-induced aberrant morphology of hippocampal dendritic spines and cognitive deficits[J]. Mol Brain, 2021, 14: 129.
[5] Zheng H, Su Y, Sun Y, et al. Echinacoside alleviates hypobaric hypoxia-induced memory impairment in C57 mice[J]. Phytother Res, 2019, 33: 1150–1160.
[6] 于艺婧,蒲玲玲,高蔚娜,等. 柑橘橘络提取物类黄酮成分分析及其抗低氧作用研究[J]. 军事医学, 2021, 45: 214–217.
[7] Liu Y, Xue C, Lu H, et al. Hypoxia causes mitochondrial dysfunction and brain memory disorder in a manner mediated by the reduction of Cirbp[J]. Sci Total Environ, 2022, 806: 151228.
[8] Alam S, Ray K, Jain V, et al. Reduced expression of Kalirin-7 contributes to working memory deficit during chronic hypobaric hypoxia exposure[J]. Behav Brain Res, 2019, 366: 135–141.
[9] 张梦雅,赵名,江亚群,等. 慢性高原低氧暴露对小鼠神经行为学的影响[J]. 中国比较医学杂志, 2023, 33: 28-35.
[10] 余孝海,刘国莹,钟明奎. 凋亡蛋白Caspase-9/Caspase-3在慢性间歇性低氧模型小鼠肝损伤中的作用[J]. 九江学院学报(自然科学版),2023,38:102–105,113.
[11] Yu Y, Li R, Pu L, et al. Citrus tangerine pith extract alleviates hypoxia-induced ileum damage in mice by modulating intestinal microbiota[J]. Food Funct, 2023, 14: 6062–6072.
[12] Xu G, Shi Y K, Sun B D, et al. DL-3-n-butylphthalide improved physical and learning and memory performance of rodents exposed to acute and chronic hypobaric hypoxia[J]. Mil Med Res, 2021, 8: 23.
[13] Cramer NP, Korotcov A, Bosomtwi A, et al. Neuronal and vascular deficits following chronic adaptation to high altitude[J]. Exp Neurol, 2019, 311: 293–304.
[14] Wang Y, Liu X J, Chen J B, et al. Citrus flavonoids and their antioxidant evaluation[J]. Crit Rev Food Sci Nutr, 2022, 62: 3833–3854.
[15] 李勃深,张宇轩,范容晖,等. 高原对线粒体功能及能量代谢影响研究进展[J]. 中国比较医学杂志, 2023, 33: 106–113.
[16] Liu G, Li Y, Liao N, et al. Energy metabolic mechanisms for high altitude sickness: Downregulation of glycolysis and upregulation of the lactic acid/amino acid-pyruvate-TCA pathways and fatty acid oxidation[J]. Sci Total Environ, 2023, 894: 164998.
[17] Wang C, Cui C, Xu P, et al. Targeting PDK2 rescues stress-induced impaired brain energy metabolism[J]. Mol Psychiatry, 2023, 28: 4138-4150.
[18] 陈怡,李晓蓉,高建,等. 丹芎方对缺氧小鼠脑组织的保护作用[J]. 中药新药与临床药理, 2008(5): 366–368.
[19] Wang X, Tang Y, Xie N, et al. Salidroside, a phenyl ethanol glycoside from Rhodiola crenulata, orchestrates hypoxic mitochondrial dynamics homeostasis by stimulating Sirt1/p53/Drp1 signaling[J]. J Ethnopharmacol, 2022, 293: 115278.
[20] Zhang J, Zhao T, Zhang P, et al. Moslosooflavone protects against brain injury induced by hypobaric hypoxic via suppressing oxidative stress, neuroinflammation, energy metabolism disorder, and apoptosis[J].J Pharm Pharmacol, 2024, 76: 44–56.
