Integrated 16S rRNA gene sequencing and LC/MS-based metabolomics ascertained synergistic influences of the combination of acupuncture and NaoMaiTong on ischemic stroke
Integrated 16S rRNA gene sequencing and LC/MS-based metabolomics ascertained synergistic influences of the combination of acupuncture and NaoMaiTong on ischemic stroke
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DOI：
10.1016/j.jep.2022.115281
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发表时间：
2022-05-04
影响因子：
5.4
通讯作者：
Wang，Shumei
中科院分区：
文献类型：
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作者：
Xian，Minghua;Shen，Lin;Wang，Shumei
民族药理相关性针灸是治疗缺血性中风的一种有效疗法，近年来在中国中得到了广泛的应用，并逐渐被更多的国家和地区接受。此外，中医药在中风的治疗中也发挥着重要的作用，其中脑脉通(NMT)是治疗中风的有效中药配方的典范。针灸和药物相结合的治疗策略被广泛应用于中风患者。然而，针药结合治疗缺血性中风的协同效应及其机制尚未完全阐明。本研究的目的是以NMT为探针，探讨针药结合治疗缺血性中风是否能产生协同作用。材料与方法采用大鼠大脑中动脉阻塞/再灌注模型，通过检测神经功能指标、脑梗塞面积、炎性因子(IL-6、IL-1β、肿瘤坏死因子-α)和氧化应激因子(超氧化物歧化酶、丙二醛)及脑源性神经营养因子的水平，评价针药结合治疗中风的疗效。随后，利用16S rRNA基因测序和LC/MS代谢组学分析，分别探讨了盲肠内容物微生态特征和血浆代谢特征。结果针刺结合NMT治疗缺血性中风的疗效明显优于单一治疗，能更有效地缩小脑梗塞面积，改善神经行为功能，减轻氧化应激和炎症反应。此外，联合疗法不仅通过丰富物种多样性、降低病原菌(如大肠杆菌)的丰度和增加有益细菌(如Turicibacter、双歧杆菌)的丰度来调整肠道微生物区系紊乱，而且还通过将代谢物血浆水平恢复到正常水平来改善代谢紊乱。相关分析结果显示肠道微生物区系与血浆代谢谱有显著相关性，尤其是细菌和异黄酮类植物雌激素代谢物的相关性较强。结论针刺结合NMT可产生协同作用，提示针药结合治疗可能更有利于缺血性中风的康复。其可能机制可能与调节肠道微生态和血浆代谢有关。细菌和异黄酮类植物雌激素代谢产物可能是针刺配合NMT治疗中风的靶点。我们目前的发现可能为治疗缺血性中风提供一种潜在的治疗策略。
Ethnopharmacological relevanceAcupuncture is an effective therapy for ischemic stroke, which has been widely used in China and gradually accepted in more countries and regions recently. In addition, Chinese medicine also plays an important role in stroke treatment, among which NaoMaiTong (NMT) is an example of an effective herbal formula for the treatment of stroke. A therapeutic strategy that combines acupuncture and medicine was widely used in stroke patients. However, the synergistic influences and mechanisms of combined acupuncture and medicine on ischemic stroke have not yet been entirely elucidated.Aim of this studyThe purpose of this study is to explore whether acupuncture and medicine combination treatments can produce synergism by using NMT, a clinically effective Chinese medicinal formula for the treatment of ischemic stroke for decades and has been demonstrated to be effective against ischemic brain injury, as a probe. Meanwhile, the potential mechanisms were investigated via cecal microbiome and plasma metabolomics to provide more strategies and basis for acupuncture-medicine combination for stroke.Materials and methodsAdopted middle-cerebral artery occlusion/reperfusion (MCAO/R) rat models, the effect for the stroke of the combination treatment consisting of acupuncture and NMT was evaluated by detecting neurological issues, cerebral infarct dimensions, levels of inflammatory factors (IL-6, IL-1β, TNF-α) and oxidative stress factors (SOD, MDA) and brain-derived neurotrophic factor (BDNF). Subsequently,16S rRNA gene sequencing and LC/MS-based metabolomic analysis were utilized to explore the characteristics of cecal-contents microecology and plasma metabolic profile, respectively. Finally, the correlation between intestinal microecological characteristics and plasma metabolic characteristics was analyzed to explore the potential mechanism of the acupuncture-NMT combination.ResultsThe efficacy of acupuncture-NMT therapy was more effective than a single treatment on ischemic stroke, with more effectively reduced infarct sizes, improved neurobehavioral deficits, and alleviated oxidative stress and inflammatory responses. Besides, the combination therapy not only adjusted gut microbiota disturbances by enriching species diversity, reducing the abundance of pathogenic bacteria (such asEscherichia-Shaigella), as well as increasing the abundance of beneficial bacteria (such asTuricibacter,Bifidobacterium), but also improved metabolic disorders by reversing metabolite plasma levels to normality. The results of the correlation analysis demonstrated a significant association between intestinal microbiota and plasma metabolic profile, especially the strong correlation ofTuricibacterand isoflavones phyto-estrogens metabolites.ConclusionThe combination of acupuncture and NMT could produce synergism, suggesting acupuncture-medicine combination therapy might be more conducive to the recovery of ischemic stroke. And the potential mechanism was probably related to the mediation of intestinal microecology and plasma metabolism.Turicibacterand isoflavones phyto-estrogens metabolites might be the targets for acupuncture-NMT combination for stroke. Our current findings could provide a potential therapeutic strategy against ischemic stroke.