Nicotinamide adenine dinucleotide treatment confers resistance to neonatal ischemia and hypoxia:effects on neurobehavioral phenotypes

Neonatal hypoxic-ischemic brain injury is the main cause of hypoxic-ischemic encephalopathy and cerebral palsy.Currently,there are few effective clinical treatments for neonatal hypoxic-ischemic brain injury.Here,we investigated the neuroprotective and molecular mechanisms of exogenous nicotinamide adenine dinucleotide,which can protect against hypoxic injury in adulthood,in a mouse model of neonatal hypoxic-ischemic brain injury.In this study,nicotinamide adenine dinucleotide(5 mg/kg)was intraperitoneally administered 30 minutes befo re surgery and every 24 hours thereafter.The results showed that nicotinamide adenine dinucleotide treatment improved body weight,brain structure,adenosine triphosphate levels,oxidative damage,neurobehavioral test outcomes,and seizure threshold in experimental mice.Tandem mass tag proteomics revealed that numerous proteins were altered after nicotinamide adenine dinucleotide treatment in hypoxic-ischemic brain injury mice.Parallel reaction monitoring and western blotting confirmed changes in the expression levels of proteins including serine(or cysteine)peptidase inhibitor,clade A,member 3N,fibronectin 1,5'-nucleotidase,cytosolic IA,microtubule associated protein 2,and complexin 2.Proteomics analyses showed that nicotinamide adenine dinucleotide ameliorated hypoxic-ischemic injury through inflammation-related signaling pathways(e.g.,nuclear factor-kappa B,mitogen-activated protein kinase,and phosphatidylinositol 3 kinase/protein kinase B).These findings suggest that nicotinamide adenine dinucleotide treatment can improve neurobehavioral phenotypes in hypoxic-ischemic brain injury mice through inflammation-related pathways.

Nicotinamide adenine dinucleotide phosphate oxidase in pancreatic diseases:Mechanisms and future perspectives

Pancreatitis and pancreatic cancer(PC)stand as the most worrisome ailments affecting the pancreas.Researchers have dedicated efforts to unraveling the mechanisms underlying these diseases,yet their true nature continues to elude their grasp.Within this realm,oxidative stress is often believed to play a causal and contributory role in the development of pancreatitis and PC.Excessive accumulation of reactive oxygen species(ROS)can cause oxidative stress,and the key enzyme responsible for inducing ROS production in cells is nicotinamide adenine dinucleotide phosphate hydrogen oxides(NOX).NOX contribute to pancreatic fibrosis and inflammation by generating ROS that injure acinar cells,activate pancreatic stellate cells,and mediate macrophage polarization.Excessive ROS production occurs during malignant transformation and pancreatic carcinogenesis,creating an oxidative microenvironment that can cause abnormal apoptosis,epithelial to mesenchymal transition and genomic instability.Therefore,understanding the role of NOX in pancreatic diseases contributes to a more in-depth exploration of the exact pathogenesis of these diseases.In this review,we aim to summarize the potential roles of NOX and its mechanism in pancreatic disorders,aiming to provide novel insights into understanding the mechanisms underlying these diseases.

Effects of different concentrations of nicotinamide on hematopoietic stem cells cultured in vitro

BACKGROUND In vitro expansion to increase numbers of hematopoietic stem cells(HSCs)in cord blood could improve clinical efficacy of this vital resource.Nicotinamide(NAM)can promote HSC expansion ex vivo,but its effect on hematopoietic stem and progenitor cells(HSPCs,CD34^(+)CD38)and functional subtypes of HSCs-shortterm repopulating HSCs(ST-HSCs,CD34^(+)CD38CD45RACD49f^(+))and long-term repopulating HSCs(LT-HSCs,CD34^(+)CD38CD45RACD49f^(+)CD90^(+))is not yet known.As a sirtuin 1(SIRT1)inhibitor,NAM participates in regulating cell adhesion,polarity,migration,proliferation,and differentiation.However,SIRT1 exhibits dual effects by promoting or inhibiting differentiation in different tissues or cells.We propose that the concentration of NAM may influence proliferation,differentiation,and SIRT1 signaling of HSCs.AIM To evaluate the effects and underlying mechanisms of action of different concentrations of NAM on HSC proliferation and differentiation.METHODS CD34^(+)cells were purified from umbilical cord blood using MacsCD34 beads,and cultured for 10-12 d in a serum-free medium supplemented with cytokines,with different concentrations of NAM added according to experimental requirements.Flow cytometry was used to detect phenotype,cell cycle distribution,and apoptosis of the cultured cells.Real-time polymerase chain reaction was used to detect the transcription levels of target genes encoding stemness-related factors,che mokines,components of hypoxia pathways,and antioxidant enzymes.Dichloro-dihydro-fluorescein diacetate probes were used to evaluate intracellular production of reactive oxygen species(ROS).Determination of the effect of different culture conditions on the balance of cytokine by cytometric bead array.RESULTS Compared with the control group,the proportion and expansion folds of HSPCs(CD34^(+)CD38)incubated with 5 mmol/L or 10 mmol/L NAM were significantly increased(all P<0.05).The ST-HSCs ratio and fold expansion of the 5 mmol/L NAM group were significantly higher than those of the control and 10 mmol/L NAM groups(all P<0.001),whereas the LT-HSCs ratio and fold expansion of the 10 mmol/L NAM group were significantly higher than those of the other two groups(all P<0.05).When the NAM concentration was>10 mmol/L,cell viability significantly decreased.In addition,compared with the 5 mmol/L NAM group,the proportion of apoptotic cells in the 10 mmol/L NAM group increased and the proportion of cells in S and G2 phase decreased.Compared with the 5 mmol/L NAM group,the HSCs incubated with 10 mmol/L NAM exhibited significantly inhibited SIRT1 expression,increased intracellular ROS content,and downregulated expression of genes encoding antioxidant enzymes(superoxide dismutase 1,peroxiredoxin 1).CONCLUSION Low concentrations(5 mmol/L)of NAM can better regulate the balance between proliferation and differentiation,thereby promoting expansion of HSCs.These findings allow adjustment of NAM concentrations according to expansion needs.

Oral Administration of Nicotinamide Mononucleotide with Bioenhancer BioPerine® Increases the Serum Concentration of Nicotinamide Adenine Dinucleotide in Healthy Human Volunteers: A Pilot, Open-Label, Cross-Over Study

Background: Bioenhancers augment the bioavailability of co-administered molecules without showing any significant effect on their own. Piperine, an alkaloid from Piper nigrum, is an established natural bioenhancer. Nicotinamide mononucleotide (NMN), an antiaging supplement, is the precursor of coenzyme nicotinamide adenine dinucleotide (NAD) that plays an important role in intracellular redox reactions. Objective: The study compared the serum concentrations of NAD in normal healthy participants, supplemented with NMN 500 mg and NMN 500 mg + 5 mg BioPerine® (95% piperine). Methods: In a randomized, open-label, crossover study, NMN (500 mg) was compared to NMN + BioPerine® (500 mg + 5 mg) in 6 healthy adults, aged 18 - 45 years. The participants received a single oral dose of NMN or NMN + BioPerine® capsules with 240 mL water, and blood samples were collected over 8hr. After a 4-day washout period, the same procedures were repeated as per the crossover design. Total NAD (NADtotal), including oxidized NAD (the oxidized) and its reduced form NADH, was measured in human serum samples. Results: The maximum concentration (Cmax) of NAD in serum was higher with NMN + BioPerine® (282 pmol/mL) compared to NMN (246 pmol/mL) alone. In the presence of BioPerine®, the NAD concentrations reached 257 pmol/mL during the first 2 hr, whereas a comparable serum concentration (246 pmol/mL) was attained only after 6 hr in NMN alone. The AUC0-8hr was 1738 pmol/mL/hr in NMN compared to 2004 pmol/mL/hr in NMN+ BioPerine®. The time to reach peak concentration (t1/2) was similar (6hr) in both groups. No clinically relevant adverse events (AE) were observed, and safety parameters remained within normal ranges in all the participants with both formulations. Conclusion: These results reveal that BioPerine® can effectively increase the NAD concentrations in the serum following NMN supplementation in healthy volunteers. The present study was registered prospectively with the Clinical Trials Registry-India (CTRI/2023/11/059982).

Research Progress on the Application of β-Nicotinamide Mononucleotides in Cosmetics

Recent studies have found that nicotinamide mononucleotide(nicotinamide mononucleotide,NMN)also has certain anti-aging,photoprotection,anti-oxidation,and soothing to the skin,while affects the level of nicotinamide adenine dinucleotide(nicotinamide adenine dinucleotide,NAD+)to improve aging-related diseases.These skincare features have laid the foundation for its application in cosmetics.NMN has been used in cosmetics and sold on the market in foreign countries,but there have been debates about its safety.It is currently as a new raw material for cosmetics in China,and its safety is also under monitoring.This paper systematically reviewed the basic properties,skin care functions and the safety of its application in cosmetics,aiming to provide theoretical reference for the development and application of NMN.

基于烟酰胺磷酸核糖转移酶(NAMPT)探讨红景天苷在非酒精性脂肪性肝病小鼠模型中的保护作用

目的研究红景天苷对非酒精性脂肪性肝病(NAFLD)的保护作用,并探讨其作用机制。方法将24只雄性KM小鼠随机分为正常组、高脂饮食(HFD)组、HFD+空白对照组、HFD+红景天苷组,每组6只,正常组小鼠予以普通饲料喂养,其余各组高脂饮食,造模14周开始给予100 mg·kg^(-1)·d^(-1)红景天苷灌胃给药,22周末取材。酶联免疫法测定血清ALT、AST、TG、TC、HDL-C、LDL-C等相关生化指标;HE染色和NAFLD活动评分(NAS)观察小鼠肝组织病理。Western Blot检测肝组织内烟酰胺磷酸核糖转移酶(NAMPT)、Sirt1、AMPKα、SREBP1的表达变化。多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果与正常组比较,HFD组小鼠肝组织存在广泛较大的脂肪滴,脂肪变性明显,NAS评分升高(P<0.01),外周血AST、ALT、TG、TC、LDL-C的含量明显增高(P值均<0.05),HDL-C含量降低(P<0.05),肝组织内NAMPT、AMPKα、Sirt1的表达降低(P值均<0.05),SERBP1的表达增加(P<0.01)。而HFD+红景天苷组较HFD组、HFD+空白对照组肝组织空泡状脂滴分布、小叶内炎症减少,肝细胞气球样变减轻,NAS评分下降(P<0.01),外周血AST、ALT、TG、LDL-C的含量明显降低(P值均<0.05),HDL-C含量升高(P<0.05),NAMPT、AMPKα、Sirt1的表达增加(P值均<0.05),SERBP1的表达减少(P<0.01)。结论红景天苷能明显改善高脂饮食诱导小鼠NAFLD的病理状态,并通过增加NAMPT、Sirt1、AMPKα的表达,减少SERBP1的表达,发挥其对NAFLD的保护作用。

NDUFS6蛋白生物信息学分析及过表达质粒的构建与鉴定

目的 应用生物信息学方法分析线粒体呼吸链复合体Ⅰ结构亚基烟酰胺腺嘌呤二核苷酸脱氢酶(泛素)铁硫蛋白6(NDUFS6)的理化性质,构建pCV702-NDUFS6过表达质粒并进行鉴定,为进一步研究NDUFS6蛋白功能奠定基础。方法 利用Expasy、UniProtKB、NCBI、SOPMA等生物信息学工具分析NDUFS6蛋白的理化性质、二级结构等;根据NDUFS6 cDNA序列构建携带NDUFS6基因的过表达质粒pCV702-NDUFS6,转染大鼠心肌细胞H9C2,并设置阴性对照(NC)组和相应空载体CON520作为阳性对照(PC)组,经嘌呤霉素筛选后,采用RT-qPCR和Western blot检测NDUFS6 mRNA和蛋白表达水平。结果 NDUFS6蛋白由116个氨基酸组成,理论等电点pI为9.37。蛋白二级结构以无规则卷曲(占50%)为主。酶切鉴定和基因测序结果显示,pCV702-NDUFS6表达质粒构建成功。RT-qPCR和Western blot结果显示,相较于NC组和PC组,过表达组NDUFS6表达水平均上调(P均<0.05)。结论 成功构建了能在心肌细胞H9C2中有效过表达NDUFS6基因的过表达质粒。

二氢槲皮素和β-烟酰胺单核苷酸顺势和逆势给药对健康大鼠生长和生理的影响

目的:通过研究二氢槲皮素和β-烟酰胺单核苷酸顺势和逆势给药对健康大鼠生长和生理的影响,进一步验证中医阴阳分药分时服用方剂学理论。方法:48只无特定病原体级SD大鼠,随机分为顺势饲喂组、逆势饲喂组、混合饲喂组、对照组各12只,雌雄各半,顺势饲喂组早晨给予二氢槲皮素溶液,下午给予β-烟酰胺单核苷酸溶液,逆势饲喂组给药顺序与顺势饲喂组相反,混合饲喂组给予二氢槲皮素和β-烟酰胺单核苷酸混合溶液灌胃,分早晚两次给药,对照组早晚给予灌胃等量超纯水,四组均每周给药5 d,共给药6周。分别于饲喂第1天、第10天、第20天、第30天、第45天时观察各组大鼠的皮肤毛发、行为状态、兴奋程度、激怒反应、睡眠状态、大便状态。分别于饲喂后第2周、第4周、第6周的周一、周三、周五测定大鼠采食量和体重,饲喂第45天检测大鼠的血液常规和血生化指标,计算脏器系数。结果:实验期间,各组大鼠的皮肤毛发、行为状态、兴奋程度、激怒反应、睡眠状态、大便状态均未见明显改变。四组雌、雄性大鼠在各个阶段的日均采食量和体重比较差异均无统计学意义(P>0.05)。给药6周后,四组大鼠肝、脾、肺、肾、胃脏器系数、白细胞计数、中性粒细胞计数、淋巴细胞计数、单核细胞计数、嗜酸性粒细胞计数、红细胞计数、血红蛋白、血清总蛋白、总胆红素、碱性磷酸酶、三酰甘油、血清总胆固醇、尿素、空腹血糖、肌酐、钾离子比较,差异均无统计学意义(P>0.05)。雄性大鼠顺势饲喂组和逆势饲喂组心脏脏器系数低于对照组(P<0.05);雌性大鼠顺势饲喂组谷丙转氨酶高于对照组(P<0.05);雄性大鼠混合饲喂组谷草转氨酶高于对照组(P<0.05);雌性大鼠混合饲喂组钠离子低于对照组,差异均有统计学意义(P<0.05)。结论:顺势给药和逆势给药可引起大鼠心脏脏器系数降低,混合给药可引起大鼠谷草转氨酶增高,钠离子降低。

烟酰胺核糖对EAE小鼠的外周抗炎作用及其机制研究

目的探讨烟酰胺核糖(NR)对实验性自身免疫性脑脊髓炎(EAE)小鼠的外周抗炎作用及机制。方法采用髓鞘少突胶质细胞糖蛋白35~55肽段(MOG35-55)诱导C57BL/6雌性小鼠制备EAE模型,随机分为EAE模型组和NR治疗组。EAE模型组每只小鼠按200μl/d剂量灌胃给予生理盐水,NR治疗组每只小鼠按500 mg/kg、200μl/d剂量灌胃给予NR,观察并记录各组小鼠临床评分和体质量。免疫后第28天处死小鼠,制备脾脏和脊髓冰冻切片,提取脊髓组织蛋白。HE染色检测外周炎性细胞浸润脊髓,免疫荧光染色检测小鼠脊髓CD4^(+)T细胞、CD68^(+)巨噬细胞数量,Western blot检测小鼠脊髓IFN-γ、IL-1β表达,免疫荧光染色检测小鼠脾脏ROCK-Ⅰ^(+)细胞、TLR4^(+)细胞、p-NF-κB^(+)细胞、TNF-α^(+)细胞、IL-1β^(+)细胞、IFN-γ^(+)细胞、IL-6^(+)细胞、IL-10^(+)细胞、IL-17^(+)细胞、i NOS^(+)细胞、Arg-1^(+)细胞数量。结果与EAE模型组相比,NR显著推迟EAE小鼠发病时间(P<0.05),降低临床评分(P<0.05或P<0.01),减轻体质量丢失,阻止外周炎性细胞浸润脊髓(P<0.01),减少脊髓CD4^(+)T细胞、CD68^(+)巨噬细胞数量(P<0.01),下调脊髓IFN-γ、IL-1β炎性因子表达(P<0.05),抑制小鼠脾脏ROCK-Ⅰ、TLR4、p-NF-κB表达(P<0.01),减少脾脏IFN-γ、i NOS、IL-6等促炎因子分泌(P<0.05或P<0.01),促进脾脏抗炎因子Arg-1、IL-10分泌(P<0.05或P<0.01)。结论NR能够有效缓解EAE小鼠临床症状,显著减轻外周和中枢神经系统炎症反应,其机制可能与抑制EAE小鼠脾脏Rho/ROCK信号通路和TLR4/NF-κB信号通路有关。

血清Nox2、ATG7水平对新生儿窒息心肌损伤的评估价值

目的探讨自噬相关基因-7(ATG7)、烟酰胺腺嘌呤二核苷酸磷酸氧化酶2(Nox2)在新生儿窒息病儿血清中的表达及早期诊断价值。方法选取2019年1月至2021年12月在郑州大学第三附属医院产科分娩的75例新生儿窒息病儿为研究组,根据是否合并心肌损伤将病儿分为窒息心肌损伤组31例,窒息非心肌损伤组44例。同期选择50例健康足月新生儿为对照组。收集一般资料,采用酶联免疫吸附测定(ELISA)测量血清ATG7水平,采用蛋白质印迹法检测Nox2水平,以Nox2/β肌动蛋白的灰度比值为Nox2表达量;采用Pearson法分析ATG7、Nox2表达的相关性及与各指标的相关性;利用受试者操作特征曲线(ROC曲线)评价Nox2、ATG7水平对新生儿窒息心肌损伤的诊断价值。结果与对照组[0.26±0.06、(4.83±0.61)ng/L]比较,新生儿窒息病儿血清Nox2(0.65±0.09)、ATG7[(21.04±3.66)ng/L]表达水平升高,且窒息心肌损伤组[0.85±0.11、(24.23±3.98)ng/L]病儿血清中Nox2、ATG7水平高于窒息非心肌损伤组[0.51±0.08、(18.80±3.43)ng/L](P<0.05);窒息心肌损伤病儿血清中Nox2、ATG7表达呈显著正相关(r=0.57,P<0.05);病儿血清中Nox2和ATG7表达与肌酸激酶同工酶(CK-MB)、缺血修饰白蛋白(IMA)、超敏C-反应蛋白(hs-CRP)、氨基末端脑钠肽前体(NT-ProBNP)、肌钙蛋白I(cTnI)、肌红蛋白表达均呈正相关(P<0.05);ROC曲线结果显示,血清Nox2、ATG7水平预测新生儿窒息合并心肌损伤的曲线下面积(AUC)及其95%CI分别为0.91(0.82,0.96)、0.89(0.80,0.95),对应的灵敏度分别为83.87%、87.10%,特异度分别为84.09%、75.00%,二者联合预测的AUC及其95%CI为0.92(0.84,0.97),灵敏度为90.32%,特异度为81.82%。结论窒息心肌损伤病儿血清中Nox2、ATG7表达均上调,二者联合检测对窒息心肌损伤有一定的诊断价值。

光甘草定烟酰胺纳米乳的制剂评价与透皮吸收

为提高光甘草定和烟酰胺的透皮吸收性能,利用两种高能乳化法制备了纳米乳(HPH-NE和HSSU-NE),对其形态、粒径、Zeta电位、稳定性和皮肤渗透性能进行了评价与考察.结果表明:两种纳米乳颗粒均为规则球形,未见聚集现象,平均粒径分别为51.60 nm和42.98 nm,Zeta电位分别为-50.44 mV和-68.50 mV,并且在离心、4℃和20℃以及加速试验条件下均具有较好的稳定性,两种方法制成的纳米乳相对普通乳液具有更好的皮肤渗透性能,光甘草定的平均渗透速率提高3~5倍,烟酰胺的平均渗透率提高1.6~2.6倍,为光甘草定的纳米制剂以及化妆品应用研究奠定了坚实基础.

烟酰胺代谢相关基因与骨关节炎的关系探索

目的·利用生物信息学方法探索骨关节炎与烟酰胺代谢相关基因之间的关系,找到具有诊断价值和治疗潜力的关键基因。方法·以“Osteoarthritis”为检索词,在GEO数据库中获取GSE12021、GSE55235和GSE55457数据集,将GSE55457作为验证集。去除GSE12021和GSE55235数据集的批次效应后,得到标准化的合并数据集,将其作为训练集,并在训练集中筛选出差异表达基因(differentially expressed genes,DEGs)。在GeneCards数据库和MSigDB数据库中获取所有烟酰胺代谢相关基因(nicotinamide metabolism-related genes,NMRGs)。将DEGs与NMRGs取交集,得到烟酰胺代谢相关差异表达基因(nicotinamide metabolism-related differentially expressed genes,NMRDEGs)。对训练集进行基因集富集分析(gene set enrichment analysis,GSEA),对NMRDEGs进行基因本体(Gene Ontology,GO)、京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析。通过LASSO(least absolute shrinkage and selection operator)和支持向量机(support vector machine,SVM)分析筛选出NMRDEGs关键基因,构建骨关节炎诊断模型,并用验证集GSE55457进行验证。通过单样本基因集富集分析(single sample gene set enrichment analysis,ssGSEA)分析免疫细胞的浸润类型。通过DGIdb数据库、ENCORI数据库和CHIPBase数据库对关键基因的mRNA进行相互作用网络和药物小分子预测。通过干扰小RNA(small interfering RNA,siRNA)敲降软骨细胞内NMRDEGs关键基因,用实时荧光定量聚合酶链反应(real-time fluorogenic quantitative polymerase chain reaction,RT-qPCR)检测关键基因敲降对软骨形成相关基因表达的影响。结果·发现了NAMPT、TIPARP等7个NMRDEGs。GO和KEGG分析富集到核因子κB信号通路和正向调节白细胞介素-1介导的信号通路等。GSEA富集到缺氧诱导因子-1转录因子通路(Hif1 Tfpathway)和多配体蛋白聚糖1(syndecan 1)通路等信号通路。LASSO分析和SVM分析共同筛选得到NPAS2、TIPARP和NAMPT关键基因并构建了骨关节炎诊断模型,验证集检验提示诊断模型诊断效果具有高准确度。ssGSEA免疫浸润分析的结果显示,巨噬细胞等15种免疫细胞存在显著差异(均P<0.05)。找到了7个针对关键基因的潜在药物小分子,19种与关键基因相互作用且上游基因与下游基因数量之和大于10的miRNA,19种与关键基因结合且上游基因与下游基因数量之和大于7的转录因子,27个聚类数>19的RNA结合蛋白。RT-qPCR结果显示,关键基因敲降会降低软骨形成相关基因的表达。结论·NPAS2、TIPARP和NAMPT为烟酰胺代谢相关的关键基因,可据此构建骨关节炎诊断模型。

烟酰胺核糖在视神经节细胞中的线粒体保护作用

目的探究烟酰胺核糖(NR)在羰基氰化物间氯苯腙(CCCP)所诱导的R28细胞氧化应激损伤模型中的保护作用。方法使用4μmol/L CCCP诱导R28细胞产生氧化应激,并使用400 nmol/L NR进行干预,CCK-8实验检测细胞活力,Annexin-V/PI双染流式细胞术检测细胞凋亡,蛋白质印记法检测细胞色素C、半胱氨酸蛋白酶-3、半胱氨酸蛋白酶-9表达水平以评估细胞凋亡;同时用四甲基罗丹明乙酯检测线粒体膜电位(MMP),MitoSOX检测线粒体活性氧(mtROS)水平,三磷酸腺苷(ATP)试剂盒检测ATP生成能力以评价线粒体功能。结果CCCP处理R28细胞后,细胞活力下降,凋亡蛋白水平和凋亡率上升,MMP下降,mtROS生成增加(P<0.05);NR预处理后,细胞活力上升,凋亡蛋白水平和细胞凋亡率下降,MMP上升,mtROS生成减少(P<0.05)。结论NR通过抑制氧化应激反应,保护线粒体功能,从而增强细胞活性,降低凋亡蛋白的表达,最终减少视网膜神经节细胞凋亡。

Impact of increasing one-carbon metabolites on traumatic brain injury outcome using pre-clinical models

Traumatic brain injury is a major cause of death and disability worldwide,affecting over 69 million individuals yearly.One-carbon metabolism has been shown to have beneficial effects after brain damage,such as ischemic stroke.However,whether increasing one-carbon metabolite vitamins impacts traumatic brain injury outcomes in patients requires more investigation.The aim of this review is to evaluate how one-carbon metabolites impact outcomes after the onset of traumatic brain injury.PubMed,Web of Science,and Google Scholar databases were searched for studies that examined the impact of B-vitamin supplementation on traumatic brain injury outcomes.The search terms included combinations of the following words:traumatic brain injury,dietary supplementation,one-carbon metabolism,and B-vitamins.The focus of each literature search was basic science data.The year of publication in the literature searches was not limited.Our analysis of the literature has shown that dietary supplementation of B-vitamins has significantly improved the functional and behavioral recove ry of animals with traumatic brain injury compared to controls.Howeve r,this improvement is dosage-dependent and is contingent upon the onset of supplementation and whether there is a sustained or continuous delive ry of vitamin supplementation post-traumatic brain injury.The details of supplementation post-traumatic brain injury need to be further investigated.Overall,we conclude that B-vitamin supplementation improves behavioral outcomes and reduces cognitive impairment post-traumatic brain injury in animal model systems.Further investigation in a clinical setting should be stro ngly considered in co njunction with current medical treatments for traumatic brain injury-affected individuals.

高效液相色谱法同时测定烟酰胺单核苷酸中5种有关物质

目的构建同时检测烟酰胺单核苷酸(nicotinamide mononucleotide,NMN)中5种有关物质[烟酰胺氧化物、三磷酸腺苷(adenosine triphosphate,ATP)、二磷酸腺苷(adenosine diphosphate,ADP)、一磷酸腺苷(adenosine monophosphate,AMP)及烟酰胺]含量的高效液相色谱法。方法采用C18色谱柱(250 mm×4.6 mm,5μm),以50 mmol/L磷酸二氢钾缓冲盐溶液(用磷酸调节pH为4.5)-甲醇为流动相进行梯度洗脱,流速为1 mL/min,进样量为20μL,检测波长210 nm,柱温20℃,试样盘控温:5℃,并对方法的专属性、灵敏性、准确性、重复性、耐用性及适用性进行验证。结果烟酰胺氧化物、ATP、ADP、AMP及烟酰胺分别在质量浓度0.4060~16.2402、0.4110~16.4403、0.4150~16.6010、0.4163~16.6507、0.4028~16.1206μg/mL范围内,质量浓度与峰面积呈现良好的线性关系,线性系数r^(2)值均大于0.998;加标回收率范围分别为98.5%~102.6%、95.6%~104.1%、98.9%~104.6%、100.0%~102.8%、100.6%~101.3%;3批样品中5种杂质含量检测结果分别为0.18%、0.20%、0.16%、0.14%、0.26%;5种杂质的检出限范围为0.04028~0.04163μg/mL,定量限范围为0.4028~0.4163μg/mL,灵敏度高;专属性、重复性和耐用性均良好。结论该方法简便易行、适用性广,适用于NMN中5种有关物质的检测,为提升其质量标准奠定基础,为食品安全提供保障。

Association of Human Whole-blood NAD+Levels with Nabothian Cyst

Objective Little is known about the association between whole-blood nicotinamide adenine dinucleotide(NAD^(+))levels and nabothian cysts.This study aimed to assess the association between NAD^(+)levels and nabothian cysts in healthy Chinese women.Methods Multivariate logistic regression analysis was performed to analyze the association between NAD^(+)levels and nabothian cysts.Results The mean age was 43.0±11.5 years,and the mean level of NAD^(+)was 31.3±5.3μmol/L.Nabothian cysts occurred in 184(27.7%)participants,with single and multiple cysts in 100(15.0%)and84(12.6%)participants,respectively.The total nabothian cyst prevalence gradually decreased from37.4%to 21.6%from Q1 to Q4 of NAD^(+)and the prevalence of single and multiple nabothian cysts also decreased across the NAD^(+)quartiles.As compared with the highest NAD^(+)quartile(≥34.4μmol/L),the adjusted odds ratios with 95%confidence interval of the NAD^(+)Q1 was 1.89(1.14–3.14)for total nabothian cysts.The risk of total and single nabothian cysts linearly decreased with increasing NAD^(+)levels,while the risk of multiple nabothian cysts decreased more rapidly at NAD^(+)levels of 28.0 to35.0μmol/L.Conclusion:Low NAD^(+)levels were associated with an increased risk of total and multiple nabothian cysts.

Cardiovascular and nonalcoholic fatty liver disease:Sharing common ground through SIRT1 pathways

As a non-communicable disease,cardiovascular disorders have become the lea-ding cause of death for men and women.Of additional concern is that cardio-vascular disease is linked to chronic comorbidity disorders that include nonal-coholic fatty liver disease(NAFLD).NAFLD,also termed metabolic-dysfunction-associated steatotic liver disease,is the greatest cause of liver disease throughout the world,increasing in prevalence concurrently with diabetes mellitus(DM),and can progress to nonalcoholic steatohepatitis that leads to cirrhosis and liver fi-brosis.Individuals with metabolic disorders,such as DM,are more than two times likely to experience cardiac disease,stroke,and liver disease that includes NAFLD when compared individuals without metabolic disorders.Interestingly,cardiovascular disorders and NAFLD share a common underlying cellular me-chanism for disease pathology,namely the silent mating type information regu-lation 2 homolog 1(SIRT1;Saccharomyces cerevisiae).SIRT1,a histone deacetylase,is linked to metabolic pathways through nicotinamide adenine dinucleotide and can offer cellular protection though multiple avenues,including trophic factors such as erythropoietin,stem cells,and AMP-activated protein kinase.Translating SIRT1 pathways into clinical care for cardiovascular and hepatic disease can offer significant hope for patients,but further insights into the complexity of SIRT1 pathways are necessary for effective treatment regimens.

Mito-specific cascade amplifier sniping metabolism homeostasis for multimodal imaging-guided antitumor bioenergetic therapy

Nicotinamide adenine dinucleotide (NAD+/NADH) pools homeostasis is recognized as an Achilles’ Heel in tumor metabolism reprogramming. However, mitochondria can enable cancer cells to overcome NADH exhaustion by providing NAD+ precursors and/or intermediates, thus promoting their survival rate and potentially driving uncontrollable proliferation. Here, a synergistic intervention NAD+/NADH homeostasis and mitochondrial metabolism strategy with magnetic resonance imaging (MRI)/photoacoustic imaging (PAI) are developed to address grand challenge of metabolic reprogramming for antitumor bioenergetic therapy. A mitochondrial-targeted cascade amplification nanoplatform ([β-MQ]TRL), triggered by NAD(P)H: quinone oxidoreductase-1 (NQO1), can enable a continuous depletion of cytosol NADH until cell death. The end-product, hydrogen peroxide (H_(2)O_(2)), can be further catalytically converted to higher toxic ·OH in proximity to mitochondria based on [β-MQ]TRL mediated Fenton-like reaction, hijacking tumorigenic energy sources and leading to mitochondrial dysfunction. Additionally, the mild thermal ablation enabled by [β-MQ]TRL further amplifies this cascade reaction to effectively prevent tumor metastasis and recurrence. This synchronous intervention strategy with MRI/PAI establishes unprecedented efficiency in antitumor bioenergetic therapy in vivo, which shows excellent promise for clinical application.

NOX4 promotes tumor progression through the MAPK-MEK1/2-ERK1/2 axis in colorectal cancer

BACKGROUND Metabolic reprogramming plays a key role in cancer progression and clinical outcomes;however,the patterns and primary regulators of metabolic reprogramming in colorectal cancer(CRC)are not well understood.AIM To explore the role of nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4)in promoting progression of CRC.METHODS We evaluated the expression and function of dysregulated and survival-related metabolic genes using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes.Consensus clustering was used to cluster CRC based on dysregulated metabolic genes.A prediction model was constructed based on survival-related metabolic genes.Sphere formation,migration,invasion,proliferation,apoptosis and clone formation was used to evaluate the biological function of NOX4 in CRC.mRNA sequencing was utilized to explore the alterations of gene expression NOX4 over-expression tumor cells.In vivo subcutaneous and lung metastasis mouse tumor model was used to explore the effect of NOX4 on tumor growth.RESULTS We comprehensively analyzed 3341 metabolic genes in CRC and identified three clusters based on dysregulated metabolic genes.Among these genes,NOX4 was highly expressed in tumor tissues and correlated with worse survival.In vitro,NOX4 overexpression induced clone formation,migration,invasion,and stemness in CRC cells.Furthermore,RNA-sequencing analysis revealed that NOX4 overexpression activated the mitogen-activated protein kinase-MEK1/2-ERK1/2 signaling pathway.Trametinib,a MEK1/2 inhibitor,abolished the NOX4-mediated tumor progression.In vivo,NOX4 overexpression promoted subcutaneous tumor growth and lung metastasis,whereas trametinib treatment can reversed the metastasis.CONCLUSION Our study comprehensively analyzed metabolic gene expression and highlighted the importance of NOX4 in promoting CRC metastasis,suggesting that trametinib could be a potential therapeutic drugs of CRC clinical therapy targeting NOX4.

β-烟酰胺单核苷酸对氧糖剥夺PC12细胞自噬的影响

为了探讨β-烟酰胺单核苷酸(β-nicotinamide mononucleotide,NMN)对氧糖剥夺(oxygen glucose deprivation,OGD)诱导的PC12细胞自噬的影响及其可能机制。通过体外建立OGD PC12细胞自噬损伤模型,MTT法检测各组细胞存活率。透射电镜观察NMN处理后的自噬小体及溶酶体,MDC荧光染色观测NMN对自噬小体的影响。Western blot检测LC3-II/LC3-I、Beclin1、p62、P-mTOR/mTOR等自噬相关蛋白表达水平。结果显示,与Con比,OGD组细胞存活率显著降低(P<0.01),自噬小体及自噬溶酶体增多(P<0.01),MDC荧光斑点及强度增强(P<0.01),Beclin1、LC3-II/LC3-I表达量显著上调(P<0.01),p62、P-mTOR/mTOR表达量显著下调(P<0.01)。与OGD组比,NMN组细胞存活率显著提高(P<0.01),NMN组和3-甲基腺嘌呤(3-methyladenine,3-MA)组自噬小体及自噬溶酶体减少(P<0.01),MDC荧光斑点及强度减弱(P<0.01),Beclin1、LC3-II/LC3-I表达量下调(P<0.01),p62、P-mTOR/mTOR表达量上调(P<0.01),而雷帕霉素(rapamycin,RAPA)组则与之相反。与RAPA组相比,RAPA+NMN组自噬小体及自噬溶酶体显著减少(P<0.01),MDC荧光斑点及强度减弱(P<0.01),Beclin1、LC3-II/LC3-I表达量显著下调(P<0.01),p62、P-mTOR/mTOR表达量显著上调(P<0.01)。因此,NMN可抑制OGD诱导的PC12细胞自噬损伤,发挥抗神经元自噬损伤的保护作用,并且这种保护作用可能与mTOR相关通路有关。本研究可为NMN防治OGD诱导的细胞自噬损伤提供一定的靶标参考,为天然化合物的开发累积一定实验室数据。