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针刺作为一种多功能神经保护疗法通过定量 iTRAQ 蛋白质组学研究改善血管性痴呆大鼠模型的认知障碍。

Acupuncture as a multifunctional neuroprotective therapy ameliorates cognitive impairment in a rat model of vascular dementia: A quantitative iTRAQ proteomics study.

机构信息

Department of Acupuncture and Moxibustion, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing, China.

School of Acupuncture-Moxibution and Tuina, Beijing University of Chinese Medicine, Beijing, China.

出版信息

CNS Neurosci Ther. 2018 Dec;24(12):1264-1274. doi: 10.1111/cns.13063. Epub 2018 Oct 2.

Abstract

AIMS

Acupuncture has been reported to affect vascular dementia through a variety of molecular mechanisms. An isobaric tag for relative and absolute quantification (iTRAQ) with high-resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses makes it possible to attain a global profile of proteins. Hence, we used an iTRAQ-LC-MS/MS strategy to unravel the underlying mechanism of acupuncture.

METHODS

Wistar rats were subjected to vascular dementia with bilateral common carotid occlusion. Acupuncture was intervened for 2 weeks at 3 days after surgery. The Morris water maze was used to assess the cognitive function. Proteins were screened by quantitative proteomics and analyzed by bioinformatic analysis. Four differentially expressed proteins (DEPs) were validated by western blot. The reactive oxygen species (ROS) production, neuron cell loss, and long-term potentiation (LTP) were determined after western blot.

RESULTS

Acupuncture at proper acupoints significantly improved cognitive function. A total of 31 proteins were considered DEPs. Gene ontology (GO) analysis showed that most of the DEPs were related to oxidative stress, apoptosis, and synaptic function, which were regarded as the major cellular processes related to acupuncture effect. Western blot results confirm the credibility of iTRAQ results. Acupuncture could decrease ROS production, increase neural cell survival, and improve LTP, which verified the three major cellular processes.

CONCLUSION

Acupuncture may serve as a promising clinical candidate for the treatment of vascular dementia via regulating oxidative stress, apoptosis, or synaptic functions.

摘要

目的

据报道,针刺通过多种分子机制影响血管性痴呆。采用相对和绝对定量同位素标记(iTRAQ)与高分辨率液相色谱-串联质谱(LC-MS/MS)分析相结合的方法,可以获得蛋白质的全局图谱。因此,我们采用 iTRAQ-LC-MS/MS 策略来揭示针刺的潜在机制。

方法

Wistar 大鼠接受双侧颈总动脉闭塞术以建立血管性痴呆模型。术后 3 天开始进行 2 周的针刺干预。采用 Morris 水迷宫评估认知功能。通过定量蛋白质组学筛选蛋白质,并进行生物信息学分析。通过 Western blot 验证 4 个差异表达蛋白(DEPs)。Western blot 后测定活性氧(ROS)生成、神经元细胞丢失和长时程增强(LTP)。

结果

针刺适当穴位可显著改善认知功能。共筛选出 31 个 DEPs。基因本体(GO)分析显示,大多数 DEPs 与氧化应激、细胞凋亡和突触功能有关,这些过程被认为是与针刺效应相关的主要细胞过程。Western blot 结果证实了 iTRAQ 结果的可信度。针刺可减少 ROS 生成、增加神经细胞存活并改善 LTP,这验证了上述三个主要的细胞过程。

结论

针刺可能通过调节氧化应激、细胞凋亡或突触功能成为治疗血管性痴呆的有前途的临床候选方法。

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