中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (25): 6463-6471.doi: 10.12307/2026.433

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葡萄籽原花青素低聚体缓解cuprizone小鼠的髓鞘脱失

王  青1，杨智超1，刘  健1，梁亚杰1，汤亿彬1，郭  羽1，宋国斌2，马存根1，2   

  1. 1山西中医药大学神经生物学研究中心，国家中医药管理局益气活血法治疗多发性硬化重点研究室，山西省晋中市   030619；2山西大同大学脑科学研究所，山西省大同市   037009
  • 收稿日期:2025-07-04 修回日期:2026-01-26 出版日期:2026-09-08 发布日期:2026-04-17
  • 通讯作者: 马存根，博士，教授，山西中医药大学神经生物学研究中心，国家中医药管理局益气活血法治疗多发性硬化重点研究室，山西省晋中市 030619；山西大同大学脑科学研究所，山西省大同市 037009
  • 作者简介:王青，女，1983年生，山西省万荣县人，汉族，2013年沈阳药科大学毕业，博士，主要从事神经免疫和神经保护的研究。
  • 基金资助:
    国家自然科学基金青年科学基金项目(81903596)，项目负责人：王青；山西省回国留学人员科研资助项目(2022-165)，项目负责人：王青；山西省中医药管理局中医药创新团队(zyytd2024039)，项目负责人：王青；山西省基础研究计划项目(202303021221162)，项目负责人：王青；山西省中医药管理局科研课题(2023ZYYB040)，项目负责人：王青；山西省卫健委2022年度中医药科研课题立项计划(2022ZYYC090)，项目负责人：马存根；山西中医药大学2022年度科技创新团队(2022TD2006)，项目负责人：王青；山西中医药大学神经炎性疾病中西医结合学科基础研究(2024XKJS-02)，项目负责人：马存根

Grape seed proanthocyanidin oligomers alleviate demyelination in cuprizone-fed mice

Wang Qing1, Yang Zhichao1, Liu Jian1, Liang Yajie1, Tang Yibin1, Guo Yu1, Song Guobin2, Ma Cungen1, 2   

  1. 1Research Center of Neurobiology, The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China; 2Institute of Brain Science, Shanxi Datong University, Datong 037009, Shanxi Province, China
  • Received:2025-07-04 Revised:2026-01-26 Online:2026-09-08 Published:2026-04-17
  • Contact: Ma Cungen, PhD, Professor, Research Center of Neurobiology, The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China; Institute of Brain Science, Shanxi Datong University, Datong 037009, Shanxi Province, China
  • About author:Wang Qing, PhD, Research Center of Neurobiology, The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China
  • Supported by:
    National Natural Science Foundation of China (Youth Fund), No. 81903596 (to WQ); Research Funding Project for Returned Overseas Chinese Scholars in Shanxi Province, No. 2022-165 (to WQ); Traditional Chinese Medicine Innovation Team of Shanxi Provincial Administration of Traditional Chinese Medicine, No. zyytd2024039 (to WQ); Basic Research Project of Shanxi Province, No. 202303021221162 (to WQ); Scientific Research Project of Shanxi Administration of Traditional Chinese Medicine, No. 2023ZYYB040 (to WQ); 2022 Traditional Chinese Medicine Scientific Research Project Plan of Shanxi Provincial Health Commission, No. 2022ZYYC090 (to MCG); 2022 Science and Technology Innovation Team of Shanxi University of Traditional Chinese Medicine, No. 2022TD2006 (to WQ); Basic Research on the Discipline of Integrated Traditional Chinese and Western Medicine for Neuritic Diseases at Shanxi University of Chinese Medicine, No. 2024XKJS-02 (to MCG)

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摘要：



文题释义：
髓鞘脱失：是指包裹在神经纤维轴突外面的髓鞘结构受到破坏或损伤，导致其功能丧失的病理过程。脱髓鞘疾病可分为中枢和外周两大类，常见的中枢神经脱髓鞘疾病有多发性硬化、视神经脊髓炎等，外周性脱髓鞘疾病包括吉兰-巴雷综合征等。
反应性星形胶质细胞：是因中枢神经系统的损伤、疾病或感染而发生形态、分子和功能重塑的星形胶质细胞，涉及到细胞转录调节的变化，以及生化、形态、代谢和生理功能的重塑，最终导致获得新功能和表型。

背景：近年来对多发性硬化症发病机制的研究提示，干预神经胶质细胞可能在减少复发和延缓残疾进展方面发挥关键作用。葡萄籽原花青素低聚体可显著抑制双环己酮草酰二腙(Cuprizone，CPZ)小鼠的髓鞘脱失。
目的：探索了葡萄籽原花青素低聚体调节星形胶质细胞保护髓鞘的作用机制。
方法：①动物实验：30只小鼠按体质量随机分为正常组、CPZ组和CPZ+低聚原花青素组，后2组每日喂养含0.2% CPZ的饲料6周诱导建立脱髓鞘模型。第5周开始正常组和CPZ组小鼠灌胃ddH2O，CPZ+低聚原花青素组小鼠灌胃葡萄籽原花青素低聚体 [50 mg/(kg·d)]，每天1次，连续给药2周。观察小鼠行为学变化，LFB、油红染色观察髓鞘病理学变化，ELISA检测脑内炎因子水平，免疫荧光染色检测脑内相关蛋白表达。②细胞实验：体外用葡萄籽原花青素低聚体(30 μg/mL)干预肿瘤坏死因子α、白细胞介素1α和C1q诱导的星形胶质细胞炎症模型，收集条件培养液后用于培养少突胶质细胞，将细胞分为正常组、模型组和模型+低聚原花青素组。采用L-乳酸脱氢酶和CCK-8检测少突胶质细胞的损伤情况和细胞活动，Western blot检测凋亡相关蛋白的表达。
结果与结论：①葡萄籽原花青素低聚体可显著改善CPZ小鼠髓鞘脱失的情况，抑制小鼠脑内促炎因子肿瘤坏死因子α、白细胞介素6、白细胞介素1α和白细胞介素17的表达，促进抗炎因子转化生长因子β的分泌，伴随着胼胝体区星形胶质细胞的增生，促炎性星形胶质细胞的标记C3d显著减少，且抑制星形胶质细胞极化相关信号分子JNK的磷酸化。②与模型组相比，葡萄籽原花青素低聚体干预后的条件培养液可显著降低炎性星形胶质细胞诱导的少突胶质细胞的凋亡，促进少突胶质细胞中Bcl-2的表达，抑制Bax和Caspase-3的表达。③结果说明，葡萄籽原花青素低聚体可抑制CPZ小鼠髓鞘脱失，通过抑制星形胶质细胞中JNK的磷酸化，减少星形胶质细胞向促炎的A1型极化，进而抑制了炎性星形胶质细胞诱导的少突胶质细胞的凋亡。
https://orcid.org/0000-0003-1284-4705 (王青) 


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 双环己酮草酰二腙, 葡萄籽原花青素低聚体, 星形胶质细胞, 炎症反应, 凋亡, 小鼠

Abstract: BACKGROUND: Recent studies on the pathogenesis of multiple sclerosis suggest that intervening in glial cells may play a key role in reducing relapses and delaying disability progression. Grape seed proanthocyanidin oligomers significantly inhibit demyelination in cuprizone-treated mice.
OBJECTIVE: To explore the mechanism by which grape seed proanthocyanidin oligomers protect myelin sheaths through regulating astrocytes. 
METHODS: (1) Animal experiment: Thirty mice were randomly divided by body mass into a normal group, a cuprizone group, and a cuprizone+oligomeric proanthocyanidins group. The latter two groups were fed a diet containing 0.2% cuprizone daily for 6 weeks to induce a demyelination model. Starting from the 5th week, mice in the normal and cuprizone groups received oral administration of ddH₂O₂, while mice in the cuprizone+oligomeric proanthocyanidins group received oral administration of grape seed proanthocyanidin oligomers [50 mg/(kg·d)] once daily for 2 consecutive weeks. Behavioral changes in mice were observed, pathological changes of myelin sheaths using LFB and oil red O staining, inflammatory factor levels in the brain were measured using ELISA, and expression of relevant proteins in the brain was detected using immunofluorescence staining. (2) Cell experiment: Grape seed proanthocyanidin oligomers (30 μg/mL) were used to intervene in an astrocyte inflammation model in vitro induced by tumor necrosis factor-α, interleukin-1α, and C1q. Conditioned medium was collected and used to culture oligodendrocytes. Cells were then divided into normal, model, and model+oligomeric proanthocyanidin groups. Oligodendrocyte injury and cell viability were assessed using L-lactate dehydrogenase and cell counting kit-8 assays, while western blot assay was used to detect apoptosis-related protein expression.
RESULTS AND CONCLUSION: (1) Grape seed proanthocyanidin oligomers significantly ameliorated myelin loss in cuprizone-treated mice by suppressing pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-6, interleukin-1α, and interleukin-17) and promoting the secretion of anti-inflammatory transforming growth factor-β. This was accompanied by the proliferation of astrocytes, a significant reduction in C3d (a marker of proinflammatory astrocytes), and suppression of JNK (a signaling molecule associated with astrocyte polarization) phosphorylation in the corpus callosum. (2) Compared with the model group, conditioned medium treated with grape seed proanthocyanidin oligomers significantly reduced apoptosis in oligodendrocytes induced by inflammatory astrocytes, promoted Bcl-2 expression, and inhibited Bax and Caspase-3 expression in oligodendrocytes. To conclude, grape seed proanthocyanidin oligomers protect against cuprizone-induced demyelination by inhibiting JNK phosphorylation in astrocytes, thereby reducing pro-inflammatory A1 polarization and subsequently inhibiting apoptosis in oligodendrocytes induced by inflammatory astrocytes.

Key words: cuprizone, grape seed proanthocyanidin oligomers, astrocytes, inflammation, apoptosis, mouse

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