高尿酸血症认知功能障碍模型大鼠皮层和海马的代谢组学研究

目的基于核磁共振氢谱(1H-NMR)代谢组学方法研究高尿酸血症诱发大鼠认知功能障碍模型的皮层和海马的内源性差异代谢物变化。方法采用含2%尿酸和2%氧嗪酸钾的高尿酸复合饲料饲喂大鼠12周,Morris水迷宫测定大鼠认知功能,并利用1H-NMR分析大鼠皮层和海马中的差异代谢物。结果高尿酸血症大鼠显示较长的逃逸潜伏期,表现出认知功能障碍;且大鼠的皮层和海马中分别筛选出27和18种差异代谢物,其中共有6种差异代谢物。γ-氨基丁酸、谷氨酸、谷氨酰胺、乳酸、甘露醇、牛磺酸和谷胱甘肽均有显著变化,可能作为高尿酸血症认知功能障碍的特征生物标志物,与学习、记忆功能损伤的发生和发展密切相关。结论高尿酸饮食可干扰机体的正常代谢,其诱发认知功能障碍机制可能与能量代谢的糖酵解和三羧酸循环、氨基酸代谢、氧化应激、神经递质转化以及细胞膜功能的紊乱等有关。     

Evolution of the Ergot Alkaloid Biosynthetic Gene Cluster Results in Divergent Mycotoxin Profiles in Claviceps purpurea Sclerotia

Research into ergot alkaloid production in major cereal cash crops is crucial for furthering our understanding of the potential toxicological impacts of Claviceps purpurea upon Canadian agriculture and to ensure consumer safety. An untargeted metabolomics approach profiling extracts of C. purpurea sclerotia from four different grain crops separated the C. purpurea strains into two distinct metabolomic classes based on ergot alkaloid content. Variances in C. purpurea alkaloid profiles were correlated to genetic differences within the lpsA gene of the ergot alkaloid biosynthetic gene cluster from previously published genomes and from newly sequenced, long-read genome assemblies of Canadian strains. Based on gene cluster composition and unique polymorphisms, we hypothesize that the alkaloid content of C. purpurea sclerotia is currently undergoing adaptation. The patterns of lpsA gene diversity described in this small subset of Canadian strains provides a remarkable framework for understanding accelerated evolution of ergot alkaloid production in Claviceps purpurea.     

Berberine ameliorates depression-like behavior in CUMS mice by activating TPH1 and inhibiting IDO1-associated with tryptophan metabolism

Berberine, which is a potential antidepressant, exhibits definite efficiency in modulating the gut microbiota. Depressive behaviors in mice induced using chronic unpredictable mild stress (CUMS) stimulation were evaluated by behavioral experiments. The markers of neurons and synapses were measured using immunohistochemical staining. An enzyme-linked immunosorbent assay was adopted to analyze serum inflammatory cytokines levels and neurotransmitters were evaluated by LC–MS/MS. Untargeted metabolomics of tryptophan metabolism was further performed using LC–MS/MS. The target enzymes of berberine involved in tryptophan metabolism were assayed using AutoDock and GRMACS softwares. Then, antibiotics was utilized to induce intestinal flora disturbance. Berberine improved the depressive behaviors of mice in a microbiota-dependent manner. Increased neurons and synaptic plasticity were observed following berberine treatment. Meanwhile, berberine decreased serum levels of TNF-α, IL-1β, and IL-4 and increased levels of IL-10. Moreover, berberine induced retraction of the abnormal neurotransmitters and metabolomics assays revealed that berberine promoted tryptophan biotransformation into serotonin and inhibited the kynurenine metabolism pathway, which was attributed to the potential agonist of tryptophan 5-hydroxylase 1 (TPH1) and inhibitor of indoleamine 2,3-dioxygenase 1 (IDO1). In conclusion, berberine improves depressive symptoms in CUMS-stimulated mice by targeting both TPH1 and IDO1, which are involved in tryptophan metabolism.     

Neuroprotective effects of Longxue Tongluo Capsule on ischemic stroke rats revealed by LC-MS/MS-based metabolomics approach

Objective: The present study aimed to evaluate the therapeutic effect and explore the underlying mechanisms of Longxue Tongluo Capsule (LTC) on ischemic stroke rats. Methods: Twenty-six rats were randomly divided into four groups, including sham group, sham + LTC group, MCAO group, and MCAO + LTC group. Ischemic stroke rats were simulated by middle cerebral artery occlusion (MCAO), and LTC treatment group were orally administrated with 300 mg/kg of LTC once daily for seven consecutive days. LTC therapy was validated in terms of neurobehavioral abnormality evaluation, cerebral infarct area, and histological assessments. The plasma metabolome comparisons amongst different groups were conducted by UHPLC-Q Exactive MS in combination with subsequent multivariate statistical analysis, aiming to finding the molecules in respond to the surgery or LTC treatment. Results: Intragastric administration of LTC significantly decreased not only the neurobehavioral abnormality scores but also the cerebral infarct area of MCAO rats. The interstitial edema, atrophy, and pyknosis of glial and neuronal cells occurred in the infarcted area, core area, and marginal area of cerebral cortex were improved after LTC treatment. A total of 13 potential biomarkers were observed, and Youden index of 11 biomarkers such as LysoPC, SM, and PE were more than 0.7, which were involved in neuroprotective process. The correlation and pathway analysis showed that LTC was beneficial to ischemic stroke rats via regulating glycerophospholipid and sphingolipid metabolism, together with nicotinate and nicotinamide metabolism. Heatmap and ternary analysis indicated the synergistic effect of carbohydrates and lipids may be induced by flavonoid intake from LTC. Conclusion: The present study could provide evidence that metabolomics, as systematic approach, revealed its capacity to evaluate the holistic efficacy of TCM, and investigate the molecular mechanism underlying the clinical treatment of LTC on ischemic stroke.     

非靶向代谢组学揭示天葵不同部位差异代谢物及其主要代谢途径

目的:利用非靶向代谢组学挖掘天葵非药用部位潜在的药用价值。方法:以天葵茎、叶、花为材料,利用超高效液相色谱-质谱法对其进行非靶向代谢组学分析。结果:天葵茎、叶、花中共鉴定出16大类101个差异代谢物(DAMs),其中包含羧酸及其衍生物、有机含氧化合物、脂质、苯及其取代衍生物、生物碱、黄酮类、萜类、苯丙素类、酚酸类、核苷酸、有机盐、内酯、有机酸、内源性植物激素、氨基酸和13个其他物质。通过聚类和京都基因与基因组百科全书(KEGG)分析发现,天葵不同部位的DAMs代谢模式存在一定的差异;其主要代谢途径为苯丙氨酸代谢途径、ABC转运途径、氨酰-tRNA合成途径及苯丙氨酸、酪氨酸和色氨酸生物合成途径。结论:通过天葵不同部位间DAMs的鉴定及其生物合成途径的分析,确定天葵茎、叶、花富含多种具有药用功效的代谢活性物质,可为天葵茎、叶、花的资源开发利用提供参考。     

醋甘遂成分群B,C对正常大鼠粪便代谢组学的研究

基于代谢组学方法,对给予醋甘遂不同成分群(B,C)前后大鼠的粪便代谢物进行比较研究,探求与醋甘遂毒性相关的差异性代谢物和代谢通路,揭示醋甘遂的毒性作用机制。采用快速液相色谱串联四极杆飞行时间质谱(UFLC-Q-TOF-MS)技术,对大鼠粪便样本进行测定,结合主成分分析(PCA)和偏最小二乘-判别分析(OPLS-DA)等多种方法筛选并鉴定与醋甘遂毒性相关的生物标志物,并采用t检验进行单变量统计分析,考察给予醋甘遂成分群B,C后正常大鼠粪便这些生物标志物的含量变化,揭示醋甘遂成分群B,C对大鼠粪便代谢组的影响程度,并结合基于MetaboAnalyst数据库的代谢通路分析探求醋甘遂成分群B,C的毒性作用机制。结果显示,与空白组相比,醋甘遂成分群B和C组大鼠粪便样本代谢组发生了明显改变,且醋B组改变程度更大,发现并鉴定了16种醋甘遂毒性潜在生物标志物及5条相关代谢通路。醋甘遂的毒性作用可能与色氨酸代谢、初级胆汁酸生物合成、氨基糖和核苷酸糖代谢、嘌呤代谢和缬氨酸,亮氨酸和异亮氨酸降解等代谢通路的紊乱有关。该研究为醋甘遂的临床安全应用提供了科学依据。     

蜘蛛香环烯醚萜部位干预CUMS致抑郁小鼠脑组织的1H-NMR代谢组学分析

目的:利用代谢组学技术分析慢性温和不可预知应激(CUMS)抑郁模型小鼠脑组织样本,寻找与抑郁相关的差异代谢物,探讨蜘蛛香环烯醚萜部位(IEFV)可能的抗抑郁作用机制。方法:42只昆明小鼠随机分为6组,包括正常组,模型组,氟西汀组(2.5 mg·kg-1),IEFV低、中、高剂量组(给药剂量分别为5.73,11.47,22.94 mg·kg-1)。采用CUMS对小鼠造模,以IEFV及阳性药(氟西汀)为干预药物,用行为指标和生化指标进行药效学评价。采用核磁共振氢谱(1H-NMR)代谢组学技术分析IEFV对CUMS抑郁模型小鼠脑组织中内源性物质的影响,结合多变量统计分析方法来确认差异代谢物,并对差异代谢物参与的代谢通路进行富集。结果:造模后,小鼠的不动时间大幅度提高、蔗糖偏好率明显下降,兴奋性神经递质5-羟色胺和去甲肾上腺素显著下降,表明造模成功。给予IEFV和氟西汀后,小鼠不动时间、蔗糖偏好率和兴奋性神经递质向正常水平回调,提示给药后抑郁状态得到了一定程度的缓解。脑组织中内源性代谢物主成分分析(PCA)结果显示,模型组可与正常组明显分开,同时IEFV低、中、高剂量组和氟西汀组均显示有偏离模型组向正常组靠拢的趋势,与行为学结果一致。模型组与正常组进行正交偏最小二乘法-判别分析(OPLS-DA)得到了16个变化显著的差异代谢物,包括12个水溶性差异代谢物和4个脂溶性差异代谢物。通过MetPA数据库分析得到7条潜在靶标代谢通路,包括三羧酸循环(TCA),牛磺酸和亚牛磺酸的代谢,丙氨酸、天冬氨酸和谷氨酸代谢等。IEFV高剂量组可显著回调11种差异代谢物。结论:IEFV可能主要通过影响能量代谢,氨基酸代谢和神经递质水平发挥抗抑郁作用,可为IEFV抗抑郁作用机制的深入研究提供参考依据。     

多组学技术在中药毒性及解毒研究中的应用

系统生物学的飞速发展为中医药深入研究提供了思路和手段,其可以全面系统地分析中药及复方多成分多靶点治疗疾病的分子机制。基于整合多组学策略研究中药毒性和解毒作用已成为中药安全性研究的方向和热点。对基因组学、转录组学、蛋白质组学、代谢组学和多组学整合技术在中药毒性及解毒作用和机制研究中的思路、方法和进展进行综述,为多组学技术在中药毒性及解毒作用深入研究中的应用提供参考。     

基于血清代谢组学技术考察通阳化浊方与四妙勇安汤治疗冠心病模型家兔的作用机制＊

目的 通过血清代谢组学技术，考察通阳化浊方与四妙勇安汤在治疗冠心病家兔中的作用机制。方法 40只雄性白兔，随机分为空白组、模型组、通阳化浊方组与四妙勇安汤组4组，每组10只。空白组正常饮食，模型组给予高脂饲料喂养至第8周，于第4周结束后，用球囊法手术损伤实验兔的右侧颈总动脉；在模型组基础上，两个实验组分别在术后，灌胃给予通阳化浊方和四妙勇安汤，连续4周，每日1次。给药结束后，模型组与实验组每组抽取3只家兔，采集全血后，用LC-MS技术进行血清代谢组学检测，运用PCA与OPLS-DA分析法来寻找差异性表达的代谢物和代谢途径。结果 相较于模型组，通阳化浊方与四妙勇安汤组分别存在45和27个差异化合物，通阳化浊方组涉及18条代谢通路，根据Impact值 > 0.05筛选出5条主要代谢通路，包括精氨酸和脯氨酸代谢、组氨酸代谢、牛磺酸和牛磺酸代谢、精氨酸生物合成和α-亚麻酸代谢；四妙勇安汤组涉及11条代谢通路，筛选出2条主要代谢通路分别为牛磺酸和牛磺酸代谢、组氨酸代谢。结论 通阳化浊方与四妙勇安汤在治疗冠心病模型家兔的过程中存在差异性代谢产物，干预相应的代谢通路可能会对药物在模型中的作用产生影响。     

基于1H-NMR植物代谢组学技术分析青海产区枸杞子的化学特征

目的:采用核磁共振氢谱(~1H-NMR)植物代谢组学技术比较青海产区枸杞子与其他产区(宁夏、甘肃、新疆、内蒙古)枸杞子的化学成分差异。方法:收集5个产区共97份枸杞子样本,其中青海61个样本,采用50%甲醇提取,检测~1H-NMR图谱,结合多元统计分析,对比青海产区枸杞子与其他产区枸杞子的化学差异性,并对各产区样本的枸杞多糖进行含量测定(以无水葡萄糖计),检测波长490 nm。结果:枸杞子的~1H-NMR图谱共检测到32个化学成分,多元统计分析表明青海产区枸杞子与其他产区样本相比,无明显分离趋势;青海产区枸杞子与宁夏产区相比,以及青海省6个不同地区的枸杞子相比,重叠样品较多,均不能显著分开。相似度结果表明,大多数样品的相似度0.85;化合物的单变量分析结果显示,除了蔗糖、葡萄糖、脯氨酸等个别代谢物在各产区样本中存在显著差异外,其余代谢物在各产区样品中的含量分布基本一致。青海与其他产区样本中枸杞多糖含量无显著性差异,且枸杞多糖含量与~1H-NMR指认的小分子化合物的相关系数处于-0.2～0.4。结论:采用~1HNMR植物代谢组学技术从整体化学组成上分析了青海产区枸杞子的化学特征,并结合枸杞多糖含量测定,显示青海产区枸杞子与其他产区枸杞子的化学差异较小。建立的基于~1H-NMR的枸杞子质量评价方法可为其质控水平提升及种植产区选择提供科学依据。