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曹娅军,王思诺,余 燕,丁妍怡,戴雅玲,杨敏光,柳维林.电针“百会”“神庭”穴对血管性认知障碍大鼠海马泛素化修饰蛋白质组学的影响[J].中国康复医学杂志,2023,(8):1025~1034
电针“百会”“神庭”穴对血管性认知障碍大鼠海马泛素化修饰蛋白质组学的影响    点此下载全文
曹娅军  王思诺  余 燕  丁妍怡  戴雅玲  杨敏光  柳维林
福建中医药大学康复医学院,福建省福州市,350122
基金项目:国家自然科学基金项目(82174478)
DOI:10.3969/j.issn.1001-1242.2023.08.001
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摘要:
      摘要 目的:探讨电针“百会”“神庭”穴对血管性认知障碍(vascular cognitive impairment,VCI)大鼠空间学习记忆和海马组织泛素化修饰蛋白质组学的影响。 方法:18只健康雄性Sprague-Dawley大鼠随机取12只结扎双侧颈总动脉制备VCI模型,6只进行假手术不结扎。术后第15天造模大鼠随机分为模型组(n=6)和电针组(n=6)。电针组电针百会、神庭穴4周。干预后用Morris水迷宫检测大鼠的空间学习记忆能力;通过泛素化修饰定量蛋白质组学鉴定泛素化蛋白质并进行生物信息学分析。 结果:水迷宫结果显示,模型组相比于假手术组逃避潜伏期延长(P<0.01),目标象限停留时间缩短(P<0.01),穿越平台次数减少(P<0.01);电针组相比于模型组逃避潜伏期缩短(P<0.01),目标象限停留时间延长(P<0.05),穿越平台次数增加(P<0.05)。泛素化修饰蛋白组学结果显示,模型组相比于假手术组有53种蛋白质上55个位点的泛素化水平上调,14种蛋白质上15个位点的泛素化水平下调(差异倍数>1.5,P<0.05);电针组相比于模型组有7种蛋白质上7个位点的泛素化水平上调,43种蛋白质上46个位点的泛素化水平下调(差异倍数>1.5,P<0.05)。其中Syt11、Usp5、CaMKIIa等8种蛋白质上8个位点的泛素化修饰水平在两个比较组中均存在差异。GO分析结果显示,两比较组中差异泛素化蛋白质多与细胞进程和生物调节过程有关,具有结合和催化活性等功能。KEGG通路富集结果显示,模型组与假手术相比,泛素化蛋白质主要富集于胞吞作用通路等17个信号通路;电针组与模型组相比,泛素化蛋白质主要富集于胆碱能突触通路等3个信号通路。 结论:电针“百会”“神庭”穴对VCI空间学习记忆的改善作用可能与海马中突触相关蛋白的泛素化修饰调控有关。
关键词:血管性认知障碍  电针  泛素化  蛋白组学
Effects of electroacupuncture Baihui and Shenting acupoints on proteomics of hippocampal ubiquitination in rats with vascular cognitive impairment    Download Fulltext
College of Rehabilitation Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian Province,350122
Fund Project:
Abstract:
      Abstract Objective: To investigate effects of electroacupuncture(EA) at Baihui(GV20) and Shenting(GV24) acupoints on spatial learning and memory and the proteomics of hippocampal tissue in rats with vascular cognitive impairment (VCI). Method: A total of 18 healthy male Sprague-Dawley rats were randomly divided into 12 rats with bilateral common carotid artery ligation to establish VCI model, and 6 rats underwent sham operation without ligation. On the 15th day after operation, the rats were randomly divided into model group (n=6) and EA group (n=6). The EA group received EA at Baihui and Shenting acupoints for 4 weeks. Morris water maze was used to test the spatial learning and memory ability of rats after intervention. The ubiquitinated proteins were identified by quantitative proteomics and bioinformatics analysis was performed. Result: Compared with the sham operation group, the water maze showed that the model group had longer escape latency (P<0.01), shorter target quadrant residence time (P<0.01), and less frequency of crossing the platform (P<0.01). Compared with the model group, the EA group had shorter escape latency (P<0.01), longer target quadrant residence time (P<0.05), and more times of crossing the platform (P<0.05). The ubiquitination modified proteomics showed that the ubiquitination levels of 55 sites on 53 proteins were up-regulated and 15 sites on 14 proteins were down-regulated in the model group compared with the sham group(fold change >1.5,P<0.05). Compared with the model group, the ubiquitination level of 7 sites on 7 proteins was up-regulated, and the ubiquitination level of 46 sites on 43 proteins was down-regulated in the EA group(fold change >1.5,P<0.05). The ubiquitination modification levels of 8 sites on 8 proteins, including Syt11, Usp5 and CaMKIIa, were different between the two comparison groups. GO analysis results showed that the difference of ubiquitinated proteins between the two groups were mostly related to cell processes and biological regulatory processes, with functions including binding and catalytic activities. KEGG pathway enrichment results showed that the ubiquitinated proteins in the EA group were mainly enriched in 17 signal pathway compared with sham operation group, and were enriched in three signaling pathways compared with the model group. Conclusion: The improvement effect of EA at Baihui and Shenting acupoints on VCI spatial learning and memory may be related to the regulation of ubiquitination of synaptic related proteins in hippocampus.
Keywords:vascular cognitive impairment  electroacupuncture  ubiquitination  proteomics
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