细胞代谢组学研究方法和应用

细胞代谢直接或间接参与细胞活动的各个方面,代谢组学技术能够检测和鉴定内源性生化反应产物,精确揭示细胞内代谢途径和过程的信息。细胞代谢组学作为代谢组学研究的一个新兴的方向,在疾病诊断、毒理学研究、药理机制和细胞培养微环境等多个领域都有所应用。     

失血性休克条件下大鼠血浆代谢组学分析

目的 应用氢核磁共振谱(1H-nuclear magnetic resonance,1H-NMR)仪对急性失血性休克条件下大鼠血浆代谢物的变化进行检测.方法 雄性Wistar大鼠10只,随机分为失血性休克组(hemorrhagicshock, HS)和对照组.测定血浆1H-NMR代谢谱,并比较两组的差异.结果 主成分分析(principal components analysis, PCA)发现两组血浆代谢物可相互区分,差异主要在于乳酸和脂类,脂类主要包括极低密度脂蛋白(very low density lipoprotein, VLDL)、低密度脂蛋白(low density lipoprotein, LDL)及不饱和脂肪酸.结论 1H-NMR分析对全面了解急性失血性休克条件下,大鼠血浆代谢组的变化具有重要的应用价值.     

血清脂联素水平与慢性乙型肝炎患者伴发代谢综合征的相关性研究

目的 研究脂联素及其受体2与慢性乙型肝炎感染伴发MS患者的肝脂肪变、纤维化和炎症的相关性.方法 收集138例慢性乙型肝炎(chronic hepatitis B,CHB)患者资料,根据患者有无代谢综合征(metabolic syndrome,MS)分为MS组(65例)和非MS组(73例),检测两组患者血清脂联素(adiponectin,Adi)水平,并比较血清酶学、糖脂类代谢相关指标水平以及HBeAg阳性率等指标是否有组间差异.结果 CHB伴发MS患者血清酶学、糖脂类代谢相关指标水平以及HBeAg阳性率明显高于非MS组.体重指数和稳态模型评估指数是CHB伴发MS患者的独立危险因素.结论 CHB患者伴发的MS与胰岛素抵抗相关,肝脏脂联素表达水平与CHB伴发MS患者疾病进展密切相关.     

代谢组学技术对牙周炎生物标志物、病理机制及与全身关系的评价

背景：近年来,代谢组学技术高速发展,研究范围十分广泛,能检测机体整体内的小分子代谢物的总体动态变化,其研究口腔疾病时可检测到机体产生的特异性代谢产物,这使得探索口腔疾病的发病机制、致病过程、早期诊断及预后监测有了新的思路。目的：对近年来代谢组学技术在牙周炎中的应用进展做一综述,同时对存在的问题及改进方法进行了展望,旨在为牙周炎诊疗提供参考依据。方法：由第一作者检索CNKI中国期刊全文数据库、万方数据库、FMRS、Web of Science及PubMed数据库2010年8月至2022年8月中收录的相关文献,中文检索词为“牙周炎,代谢组学,唾液,龈沟液”,英文检索词为“metabolomics,metabonomics,periodontitis,saliva,gingival sulcus fluid”。首先简要阅读文章摘要及主要内容,对文献进行初步筛选,将与文章主题不相关的文献排除,最终纳入的文献数为52篇。结果与结论：目前牙周炎的代谢组学研究样本来源主要为唾液、龈沟液、血清、细菌、尿液、细胞和组织,不同的样本来源均有一定的优劣。代谢组学技术与牙周炎的相关研究结果主要集中在蛋白质、脂...     

糖尿病肾病小鼠CD4+T细胞的脂质组学研究

目的：初步探讨糖尿病肾病(DKD)小鼠CD4⁺T免疫细胞在脂质组学方面的差异,筛选出具有生物学意义的差异代谢产物。方法：先采用CD4 (L3T4) MicroBeads免疫磁珠法分离BKS.Cg-Dock7^m+/+Lepr^db/J自发性DKD小鼠脾脏CD4⁺T免疫细胞;流式细胞术鉴定CD4⁺T免疫细胞纯度,LC-MS/MS技术检测CD4⁺T免疫细胞非靶向脂质组学,对差异代谢产物进行分析。结果：LC-MS法检测出463个代谢产物;PCA和OPLS-DA分析显示代谢组分明显分离;筛选出24种差异代谢物。KEGG及富集分析可知差异代谢物涉及甘油磷脂代谢紊乱。结论：CD4⁺T细胞磷脂类代谢与DKD的发生密切相关,靶向DKD CD4⁺T细胞的磷脂类代谢可能是DKD治疗的新方向。     

基于代谢组学的干细胞研究进展

代谢组学是一种定性和定量分析复杂生物样品中所有小分子代谢物的组学方法,主要考察生物体系受刺激或扰动后内源性代谢物的变化,是系统生物学的重要组成部分.近年来代谢组学在评判细胞功能状态及决定细胞命运的代谢途径的研究中取得了很大的进展.干细胞治疗作为一种再生医学,为各种退行性疾病的治疗带来了新的希望,成为目前的研究热点,本文重点综述了代谢组学在分析各种干细胞研究中的进展.     

代谢组学及其在中医药领域的应用概况

代谢组学技术是近年来从整体角度发展起来的利用机体代谢终末产物研究机体内在变化及外在刺激作用的一种新兴技术,其研究思路与中医整体观近似,对阐明中医理论、中医的诊疗原则等具有积极意义。     

多组学技术在人子宫内膜容受性中的研究进展

子宫内膜容受性是胚胎成功着床的关键,其异常是胚胎着床失败及不孕的主要原因,而传统的子宫内膜形态学观察方法无法充分确定子宫内膜容受性的状态。近年来,多组学技术的发展为生理病理条件下的子宫内膜功能特征研究提供了便捷有效的分析方法。因此,本文对转录组学、蛋白质组学及代谢组学等在人类子宫内膜容受性相关研究中的应用进行综述,旨在深入了解子宫内膜容受性并发掘新技术以强化子宫内膜植入窗口期检测的准确性,从而提高辅助生殖技术周期中怀孕的几率。     

创伤性脊髓损伤患者血清和尿液的代谢组学分析

背景:创伤性脊髓损伤在临床上主要依赖于量表评估与影像学检查,但对于损伤程度的预后评估具有一定局限性,利用代谢组学技术进行生物标志物筛选,对于估计病变范围、损伤与恢复程度以及开发新疗法具有重要意义。目的:使用代谢组学技术来表征创伤性脊髓损伤患者的代谢特征,探寻潜在的生物标志物及失调的代谢途径。方法:收集20例创伤性脊髓损伤患者(观察组)和10例健康受试者(对照组)的血清和尿液样本,进行代谢物分析,然后利用多元变量统计分析进行数据处理,筛选差异代谢物。通过MetaboAnalyst软件进行代谢通路富集,应用logistic回归构建生物标志物组合模型,并分析其与美国脊髓损伤协会(ASIA)分级的关系。结果与结论:两组受试者的血清和尿液中分别检测出160种和73种具有显著差异的代谢物。通路富集分析显示,创伤性脊髓损伤后脂质代谢出现明显的紊乱,包括鞘脂类、亚油酸、α-亚麻酸和花生四烯酸代谢以及糖基磷脂酰肌醇生物合成。识别出他索沙坦和葫芦素糖苷这组生物标志物,并且二者构成的代谢物组合在血清和尿液中的水平与ASIA分级存在相关性。由此可见,代谢组学技术为进一步理解创伤性脊髓损伤病理机制、筛选治疗靶点提供帮助。识别出的代谢生物标志物组合可能为评估创伤性脊髓损伤的严重程度提供参考。     

家兔心脏停搏自主循环恢复后血清代谢组学特征分析

目的:探讨分析家兔心脏停搏(CA)自主循环恢复(ROSC)后血清代谢组学特征变化.方法:10只雄性新西兰家兔,采用窒息法制备CA家兔模型,于CA前、ROSC后15 min、3 h、6 h和24 h各收集2 mL血液样品,通过气相色谱-质谱联用(GC-MS)测定各时间点血清代谢产物,采用Simca P软件对血清代谢轮廓特...     

Resolvins and protectins in the termination program of acute inflammation

The physiological resolution of a well-orchestrated inflammatory response is essential to maintain homeostasis. Therefore, gaining a comprehensive understanding in molecular terms of the events that direct the termination of acute inflammation is imperative. Recently, new families of local-acting mediators were discovered that are biosynthesized from the essential fatty acids eicosapentaenoic acid and docosahexaenoic acid. These new chemical mediators are endogenously generated in inflammatory exudates collected during the resolution phase, and were termed resolvins and protectins because specific members of these families control the magnitude and duration of inflammation in animals. In addition, recent results indicate novel actions of resolvins and protectins in removing chemokines ferried from the tissue by apoptotic neutrophils and T cells during resolution. Here, we review recent advances on the biosynthesis and actions of these novel anti-inflammatory and proresolving mediators.     

HLA-DR expression,cytokines and bioactive lipids in sepsis

Sepsis accounts for more than 200,000 deaths annually in the USA alone. Both inflammatory and anti-inflammatory responses occur simultaneously in sepsis, the early phase dominated by the hyperinflammatory response and the late phase by immunosuppression. This late immunosuppression phase leads to loss of the delayed type hypersensitivity response, failure to clear the primary infection and development of secondary infections. Based on the available data, I hypothesize that failure to produce adequate amounts of inflammation resolving lipid mediators may be at the centre of both the hyperinflammatory response and late immunosuppression seen in sepsis. These proresolving lipids – lipoxins, resolvins and protectins – suppress exacerbated activation of leukocytes and macrophages, inhibit excess production of pro-inflammatory cytokines, initiate resolution of inappropriate inflammation, augment clearance of bacteria and other pathogens, and restore homeostasis. If true, this implies that administration of naturally occurring lipoxins, resolvins, protectins, maresins and nitrolipids by themselves or their more stable synthetic analogues such as 15-epi-16-(para-fluorophenoxy)-lipoxin A₄-methyl ester, a synthetic analogue of 15-epi-lipoxin A₄, and 15(R/S)-methyl-LXA₄ may form a new approach in the prevention (in the high-risk subjects), management of sepsis and in resolving the imbalanced inflammatory process such that sepsis is ameliorated early. In addition, recent studies have suggested that nociceptin and cold inducible RNA binding protein (CIRBP) also have a role in the pathobiology of sepsis. It is suggested that both nociceptin and CIRBP inhibit the production of lipoxins, resolvins, protectins, maresins, and nitrolipids and thus play a role in sepsis and septic shock.     

Resolution of inflammation: the beginning programs the end

Acute inflammation normally resolves by mechanisms that have remained somewhat elusive. Emerging evidence now suggests that an active, coordinated program of resolution initiates in the first few hours after an inflammatory response begins. After entering tissues, granulocytes promote the switch of arachidonic acid-derived prostaglandins and leukotrienes to lipoxins, which initiate the termination sequence. Neutrophil recruitment thus ceases and programmed death by apoptosis is engaged. These events coincide with the biosynthesis, from omega-3 polyunsaturated fatty acids, of resolvins and protectins, which critically shorten the period of neutrophil infiltration by initiating apoptosis. Consequently, apoptotic neutrophils undergo phagocytosis by macrophages, leading to neutrophil clearance and release of anti-inflammatory and reparative cytokines such as transforming growth factor-beta1. The anti-inflammatory program ends with the departure of macrophages through the lymphatics. Understanding these and further details of the mechanism required for inflammation resolution may underpin the development of drugs that can resolve inflammatory processes in directed and controlled ways.     

Endogenous pro-resolving and anti-inflammatory lipid mediators: the new hope of atherosclerotic diseases

Atherosclerosis is a complex disease process in which genetic, lipid, cellular, and immunological factors combine to determine the location, severity, and timing of lesion development and clinical events. It has been demonstrated, however, that inflammation governed atherosclerosis during the course of development of atherosclerosis. It has also been demonstrated to be effective to decrease the cardiovascular events and improve the prognosis of atherosclerotic diseases by regulating inflammatory reaction (e.g., statins). However, endogenous mechanisms of limiting inflammation in atherosclerosis are still unclear. Recent studies showed that lipoxidase/leukotrienes (LOX/LTs) pathway played important role in the ignition and development of atherosclerosis, whereas resolvins (E-series resolvins and D-series resolvins) and protectins [protectin D1 (PD1) and neuroprotectin D1 (NPD1)], endogenous lipid-derived mediators, inhibited inflammation through pro-resolution and counter-modulating immune inflammation reaction in atherosclerosis. Hence, we hypothesize that increased endogenous lipid mediators mentioned above play a vital role in anti-atherosclerosis and plaque stabilization through pro-resolution and anti-inflammation by LOX/LTs pathway. In addition, we predict that the endogenous lipid mediators may be a new target for treatment of atherosclerotic diseases.     

Annexin A1 based inflammation resolving mediators and nanomedicines for inflammatory bowel disease therapy

Inflammatory bowel diseases (IBD) such as Crohn’s Disease (CD) and Ulcerative Colitis (UC) are chronic, progressive, and relapsing disorders of the gastrointestinal tract (GIT), characterised by intestinal epithelial injury and inflammation. Current research shows that in addition to traditional anti-inflammatory therapy, resolution of inflammation and repair of the epithelial barrier are key biological requirements in combating IBD. Resolution mediators include endogenous lipids that are generated during inflammation, e.g., lipoxins, resolvins, protectins, maresins; and proteins such as Annexin A1 (ANXA1). Nanoparticles can specifically deliver these potent inflammation resolving mediators in a spatiotemporal manner to IBD lesions, effectively resolve inflammation, and promote a return to homoeostasis with minimal collateral damage. We discuss these exciting and timely concepts in this review.     

The resolution code of acute inflammation: Novel pro-resolving lipid mediators in resolution

Studies into the mechanisms in resolution of self-limited inflammation and acute reperfusion injury have uncovered a new genus of pro-resolving lipid mediators coined specialized pro-resolving mediators (SPM) including lipoxins, resolvins, protectins and maresins that are each temporally produced by resolving-exudates with distinct actions for return to homeostasis. SPM evoke potent anti-inflammatory and novel pro-resolving mechanisms as well as enhance microbial clearance. While born in inflammation-resolution, SPM are conserved structures with functions discovered in microbial defense, pain, organ protection and tissue regeneration, wound healing, cancer, reproduction, and neurobiology-cognition. This review covers these SPM mechanisms and other new omega-3 PUFA pathways that open their path for functions in resolution physiology.     

Does aspirin-induced oxidative stress cause asthma exacerbation?

Aspirin-induced asthma (AIA) is a distinct clinical syndrome characterized by severe asthma exacerbations after ingestion of aspirin or other non-steroidal anti-inflammatory drugs. The exact pathomechanism of AIA remains unknown, though ongoing research has shed some light. Recently, more and more attention has been focused on the role of aspirin in the induction of oxidative stress, especially in cancer cell systems. However, it has not excluded the similar action of aspirin in other inflammatory disorders such as asthma. Moreover, increased levels of 8-isoprostanes, reliable biomarkers of oxidative stress in expired breath condensate in steroid-naïve patients with AIA compared to AIA patients treated with steroids and healthy volunteers, has been observed. This review is an attempt to cover aspirin-induced oxidative stress action in AIA and to suggest a possible related pathomechanism.     

Accumulation of Eicosapentaenoic Acid in Membranes of Erythrocytes and Leukocytes during Alterative-Exudative and Allergic Inflammation

Children patients with alterative-exudative (gastrointestinal diseases) and allergic inflammation (bronchial asthma) exhibited similar changes in fatty acide composition of erythrocyte and leukocyte membranes. They included accumulation of considerable amounts of eicosapentaenoic acid and decrease in the content of docosahexaenoic acid (long-chain derivative of eicosapentaenoic acid). The accumulation of eicosapentaenoic acid and decrease in the content of docosahexaenoic acid in cell membranes probably play a role in the pathogenesis of chronic inflammatory diseases. It is related to a possible decrease in the formation of antiinflammatory eicosanoids (series-5 leukotrienes) and protective compounds (resolvins and protectins) from fatty acids and changes in the physicochemical properties of cell membranes.     

Is sepsis a pro-resolution deficiency disorder?

Sepsis is due to a systemic inflammatory response to both infectious and non-infectious disorders; and when it leads to hypotension and organ dysfunction, septic shock occurs. Mortality in sepsis is due to multiple organ dysfunction. The early stages of sepsis are characterized by excessive generation of inflammatory mediators; however, as sepsis develops into chronic severe sepsis, immunosuppression dominates. Despite several advances in our understanding of the pathogenesis of sepsis both its prevention and management remains elusive. It is proposed that sepsis is due to failure of production of appropriate amounts of pro-resolution bioactive lipids such as lipoxins, resolvins, protectins, maresins and nitrolipids that suppress inappropriate inflammation, production of pro-inflammatory cytokines, free radical generation, and leukocyte activation and enhance resolution of inflammation and wound healing.     

Novel Lipid Mediators and Resolution Mechanisms in Acute Inflammation: To Resolve or Not?

Because inflammation is appreciated as a unifying basis of many widely occurring diseases, the mechanisms involved in its natural resolution are of considerable interest. Using contained, self-limited inflammatory exudates and a systems approach, novel lipid-derived mediators and pathways were uncovered in the resolution of inflammatory exudates. These new families of local mediators control both the duration and magnitude of acute inflammation as well as the return of the site to homeostasis in the process of catabasis. This new genus of specialized proresolving mediators (SPM) includes essential fatty acid–derived lipoxins, resolvins, protectins, and, most recently, maresins. These families were named based on their unique structures and potent stereoselective actions. The temporally initiated biosynthesis of SPM and their direct impact on leukocyte trafficking and macrophage-directed clearance mechanisms provide clear evidence that resolution is an active, programmed response at the tissue level. Moreover, SPM that possess anti-inflammatory (ie, limiting PMN infiltration) and proresolving (enhance macrophage uptake and clearance of apoptotic PMN and microbial particles) actions as well as stimulating mucosal antimicrobial responses demonstrate that anti-inflammation and proresolution are different responses of the host and novel defining properties of these molecules. The mapping of new resolution circuits has opened the possibility for understanding mechanisms that lead from acute to chronic inflammation, or to the resolution thereof, as well as to potential, resolution-based immunopharmacological therapies.