纳米聚苯乙烯长期暴露中小鼠代谢谱的动态变化

目的  基于代谢组学的方法,揭示纳米塑料长期暴露对机体代谢的动态影响。方法  13月龄的C57BL/6J雄鼠以0、1、10 mg/kg剂量的纳米聚苯乙烯(polystyrene nanoparticles, PS-NPs)进行灌胃,灌胃暴露时长为9个月。分别于暴露第1、3、6、9个月,处死部分小鼠并收集其血液。最后采用TM广靶代谢组学技术对血清差异代谢物进行分析。结果  同剂量组内不同暴露时间节点(暴露第1、3、6、9个月)的样本点均能明显分离,发现尿胆素含量随暴露时长的增加而减少,并与卟啉与叶绿素代谢通路密切相关。在长期暴露结束后(暴露第9个月),在对照组与不同暴露剂量组间比较中共发现135个差异代谢物。其中50个代谢物含量上调,85个代谢物含量下调。聚类分析发现,花生四烯酸代谢和嘌呤代谢通路是主要的受干扰通路,与该通路相关的差异代谢物含量(2,3-dinor-8异前列腺素F2α、黄嘌呤、2-脱氧肌苷、腺苷-3′-磷酸、次黄嘌呤等)均明显上调。结论  小鼠在长期的PS-NPs暴露过程(中年-老年)中,其血清代谢谱发生改变。其中,尿胆素与PS-NPs暴露时长相关,是潜在的敏感标志物。2,3-dinor-8异前列腺素F2α、黄嘌呤等是潜在的特异性标志物,这些代谢物提示花生四烯酸代谢和嘌呤代谢通路是研究PS-NPs暴露损伤的可能机制通路。     

基于高效液相色谱-质谱技术的宫颈上皮内瘤样变患者血液代谢组学分析

目的:利用代谢组学的研究方法,对宫颈上皮瘤样变(CIN)患者血浆中可能的相关标志物进行筛选。方法:运用高效液相色谱-质谱技术(HPLC-MS)对31例CIN患者(CIN组)和33例健康人(对照组)的血浆进行检测,结合主成分分析法(PCA)对差异代谢物进行模式识别分析并观察组间含量变化。结果:CIN组和对照组的血浆代谢谱得到明显分离,发现并鉴定了10个与CIN相关的潜在生物标志物。与对照组相比,CIN患者血浆中组氨酸、谷氨酸、酪氨酸、丙氨酸、牛磺酸、肌酸、肌醇、硬脂酸和花生四烯酸的含量减少,差异有统计学意义(P<0.05),色氨酸含量显著增多,差异有统计学意义(P<0.05)。结论:基于HPLC-MS结合PCA的代谢组学技术能有效地区分CIN患者和健康人的血浆代谢物,为宫颈癌的早期诊断提供新的途径。     

长期睡眠剥夺下调AMPK/SIRT1/PGC-1α通路引起小鼠脂代谢紊乱

目的探讨长期睡眠剥夺对小鼠肝组织腺苷酸激活蛋白激酶(adenosine 5’-monophosphateactivated protein kinase,AMPK)/沉默信息调节因子1(silent information regulation1,SIRT1)/过氧化物酶体增殖物激活受体γ共激活因子-1α(peroxisome proliferator-activated receptorγcoactivator 1α,PGC-1α)系统的影响。方法2月龄C57BL/6J小鼠16只随机分为睡眠剥夺组和正常对照组,睡眠剥夺组利用睡眠剥夺仪每天剥夺睡眠20 h,每周剥夺6 d,连续10周。正常对照组在相同实验条件下自由睡眠10周。每周测量一次体重,10周后进行小鼠体成分测定,并采集血液、肝脏和脂肪组织样本,检测血清褪黑素水平、肝组织AMPKα、SIRT1、PGC-1α及胆固醇调节元件结合蛋白1C(sterol regulatory element-binding protein 1C,SREBP-1C)蛋白表达水平,以及脂肪组织中脂肪合成关键酶脂肪酸合成酶(fatty acid synthase,FAS)、乙酰辅酶A羧化酶1(acetyl CoA carbosylase 1,ACC1)表达水平。结果建模结束后,睡眠剥夺组小鼠体重显著高于对照组(F=10.955,P=0.006),且从第6周末开始,睡眠剥夺组小鼠体重明显高于对照组(F=8.535,P=0.012),并且睡眠剥夺组小鼠体脂量[(14.58±1.70)%]高于对照组[(11.24±1.64)%](t=4.007,P=0.001),血清褪黑素浓度[(2.33±0.53)ng/ml]低于对照组[(2.87±0.23)ng/ml](t=2.643,P=0.019),AMPK通路中的AMPKα(t=7.134,P<0.001)、SIRT1(t=7.531,P<0.001)、PGC-1α(t=11.537,P<0.001)表达水平降低,AMPK/SIRT1/PGC-1α通路下调。而SREBP-1C表达水平升高,差异有统计学意义(t=-4.496,P=0.001)。同时睡眠剥夺小鼠脂肪组织中FAS(t=-4.375,P=0.001)、ACC1表达水平均升高(t=-4.072,P=0.001)。结论长期睡眠剥夺可致褪黑素受体介导的AMPK/SIRT1/PGC-1α通路抑制,并且脂肪合成相关基因表达增强,引起能量代谢特别是脂代谢稳态失调和体脂增加。     

黄药子中毒导致肝损伤的机制研究

目的分析中药黄药子水提取物诱导小鼠肝损伤后的基因表达谱,探讨黄药子中毒导致肝损伤的分子作用机制。 方法SPF级雌性昆明小鼠40只,按随机数字表法分为4组,每组10只。黄药子干预组小鼠分别以低(6 g/kg/d)、中(12 g/kg/d)、高(24 g/kg/d)剂量的黄药子水提取物连续灌胃21 d,正常对照组以同体积纯水灌胃。对比观察4组小鼠的体重增长值、肝脏指数、血清丙氨酸氨基转移酶(ALT)和天冬氨酸转移酶(AST)活性以及肝组织经苏木精-伊红(HE)染色后的病理变化,确定可引起小鼠明显肝损伤的给药剂量。利用RNA-Seq技术分别构建黄药子诱导组(DB组)和对照组(Normal组)小鼠肝脏的数字化基因表达谱(DGE)文库,对比两个DGE文库的差异表达基因,并进行基因本体论(GO)功能注释和京都基因与基因组百科全书(KEGG)通路分析。 结果与对照组相比,仅高剂量组小鼠出现明显的肝损伤,表现为同时存在体重增长值降低、肝脏指数增高、ALT及AST活性增高(P均<0.05)以及肝组织病理学变化(结构明显破坏、肝索受压紊乱、肝细胞明显肿胀且呈气球样变性、肝小叶内中性粒细胞浸润)。以高剂量组小鼠为DB诱导组,对照组为Normal组,分别测序并构建肝组织的DGE文库：两组分别检测出13 214 893和11 124 617条碱基序列,过滤并剔除带有接头、带有N碱基和测序质量低的reads后分别得到12 819 933和10 786 300条序列。根据所得的两组DEG文库,对比筛选出了312个显著差异表达的基因,其中上调基因148个,下调基因164个。对两组差异表达基因进行聚类分析：两组均表达的高质量序列有9537个,仅在DB组表达的有702个,仅在Nomal组表达的有539个。GO功能注释主要归类为生物学过程和分子功能两个条目。根据KEGG通路分析结果,差异表达的基因主要涉及8条通路：视黄醇代谢通路、花生四烯酸代谢通路、过氧化物酶体增殖物激活受体(PPARs)信号通路、细胞色素P450(CYP450)酶代谢通路、CYP450酶催化的外源物质代谢通路、甾类激素生物合成通路、谷胱甘肽代谢通路、不饱和脂肪酸的生物合成过程。 结论黄药子的诱导作用严重影响了小鼠肝脏的结构、功能以及相关功能基因的表达;其造成肝损伤的作用机制可能是黄药子在肝内经CYP450酶代谢产生了带有自由基的代谢产物,使胞质膜和细胞器脂质过氧化、机体抗氧化能力减弱,或通过直接影响肝内脂质代谢、增强肝脏脂肪酸氧化反应,从而导致肝损伤。     

雄黄与亚砷酸钠亚急性暴露对小鼠砷代谢及氧化应激影响的比较

目的探讨经口染毒雄黄、亚砷酸钠后,小鼠体内砷代谢及氧化应激指标的变化,为评价雄黄的毒性提供实验数据。方法清洁级雌性昆明小鼠随机分为对照组(0.5%羧甲基纤维素钠)、0.5 g/kg雄黄组和0.1 g/kg亚砷酸钠组,各组均灌胃染毒3 d,用氢化物发生-超低温捕集-原子吸收分光光度法分别测定尿液和肝组织中无机砷(iAs)、一甲基胂(MMA)和二甲基胂(DMA)。用试剂盒法测定全血中GSH和肝脏T-AOC水平。结果雄黄组小鼠尿液和肝脏中各形态砷和总砷含量高于对照组,但低于亚砷酸钠组;一甲基化率(FMR)和二甲基化率(SMR)高于亚砷酸钠组,差异均有统计学意义(P<0.05)。雄黄组全血GSH和肝脏T-AOC水平均高于亚砷酸钠组(P<0.05),其全血GSH和肝脏T-AOC水平与对照组相比差异均无统计学意义(P>0.05)。结论与亚砷酸钠相比,雄黄中的砷在小鼠体内代谢较慢,对氧化应激指标影响较小。     

慢性砷中毒患者头发代谢物特征的代谢组学研究

[目的]分析和对比慢性砷中毒患者与健康对照者的头发样本,筛选差异代谢物,分析相关代谢途径,为砷毒性机制研究提供理论依据。[方法]采用气相色谱/质谱联用法(GC/MS)和代谢组学方法,研究5名慢性砷中毒患者(观察组)和5名健康者(对照组)的头发样本的代谢物差异。利用主成分分析法(PCA)和偏最小二乘判别分析(PLS-DA)分析两组的代谢图谱差异,找出差异代谢物。[结果]慢性砷中毒患者尿砷[(1.32±0.09)μmol/L]明显高于砷中毒标准(1.17μmol/L)和对照组[(0.29±0.10)μmol/L](P0.001)。共检出10种差异代谢物,在慢性砷中毒患者中有4种上调,6种下调。代谢通路分析筛选出4种有意义的差异代谢物,与对照组相比,在观察组中分别为苯乙醛和γ-氨基丁酸含量升高,而谷胱甘肽和丙氨酸含量降低。这4种代谢物均与机体氧化应激相关。[结论]验证了砷的慢性毒性机制主要涉及氧化应激,同时为探索氧化应激的发生机制提供了可能的思路和线索。     

双绒毛膜双胎生长不一致的非靶向代谢组学研究

目的 利用非靶向代谢组学技术探索双绒毛膜双胎妊娠生长不一致的两胎儿脐血的代谢轮廓差异。方法 纳入26例符合双绒毛膜双胎生长不一致诊断标准的孕妇,利用超高效液相色谱-质谱联用技术(UPLC-MS)对双胎胎儿脐血进行代谢物检测,通过偏最小二乘判别分析(PLS-DA)对质谱数据进行多元变量模式分析,并根据PLS-DA模型计算得到变量权重值(VIP),结合单变量统计分析Wilcoxon秩和检验、差异倍数(fold change, FC)分析,以VIP>1,FC>1.2或FC<0.83作为筛选条件筛选差异代谢物特征,并进行通路分析。结果 双胎生长不一致中一胎儿的体重小于同胎龄儿的第10百分位数,提示该胎儿存在生长受限的情况。两胎儿在相同母体环境下,生长受限儿与正常胎儿脐血代谢物特征存在差异。两组对比最终共鉴定出35个差异代谢物。其中蛋氨酸、反式-2-十二碳酰肉碱、17α,21-二羟基孕烯醇酮、前列腺素D3等10个代谢物在生长受限儿中上调,胆碱、精氨酸、硫胱醚等25个代谢物下降。通路分析显示鉴定出的差异代谢物归属于蛋氨酸代谢,精胺代谢,甜菜碱代谢、甘油磷脂代谢等通路。结论 双绒毛...     

睡眠剥夺对大鼠肝功能的影响

目的探究急性睡眠剥夺对Wistar大鼠肝脏功能的影响。方法 30只Wistar大鼠随机分为3组,每组10只,雌雄各半,分别为睡眠剥夺组、假暴露组、正常笼养组,睡眠剥夺组连续剥夺睡眠6 d。6 d后对其肝脏进行称重、形态学观察,测定血清中天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)活力和总蛋白(TP)、白蛋白(ALB)含量。结果睡眠剥夺组大鼠血清中的AST、ALT的活力分别高于假暴露组及正常笼养组,差异有统计学意义(P0.01)。光镜下,睡眠剥夺组大鼠的肝细胞可见充血坏死,肝小叶间有大量炎细胞浸润。结论睡眠剥夺能够引起Wistar大鼠肝脏急性损伤。     

大鼠膝骨关节炎潜在生物标志物及代谢通路筛选

目的:筛选与大鼠膝骨关节炎相关的差异代谢物及其代谢通路,为深入研究骨关节炎生物标志物提供线索。方法:60只雄性SPF级SD大鼠按体质量（300 ~ 350 g）采用随机数字表法分为模型组和对照组,每组30只。实验周期分别为4、8和12周,每个周期每组10只大鼠。模型组大鼠左侧膝关节采用改良Hulth法行造模手术,5 d...     

乙肝肝硬化伴前肌少症患者的代谢组学双平台研究

目的通过代谢组学双平台分析, 揭示乙型肝炎(简称乙肝)肝硬化伴前肌少症患者的代谢特征, 识别潜在的生物标志物, 为营养支持治疗提供方向。方法征集青岛大学附属医院乙肝肝硬化住院患者的临床资料和血样本。通过CT测量第三腰椎横截面的骨骼肌面积, 计算骨骼肌指数。前肌少症的诊断标准:男性骨骼肌指数<46.96, 女性骨骼肌指数<32.46。最终纳入15例乙肝肝硬化伴前肌少症的患者和14例乙肝肝硬化非前肌少症的患者。通过液相色谱-质谱和气相色谱-质谱对血浆进行代谢组学分析。结果液相色谱-质谱和气相色谱-质谱分别富集到5种和28种差异代谢通路, 其中氨基酸相关的代谢通路、尿素循环通路、乙醛酸盐和二羧酸盐代谢通路与乙肝肝硬化肌肉质量减少相关。3-羟基丙醛、苯丙酸、甜菜碱醛、(r)-3-羟基丁酸、磷酸羟基丙酮酸和肌酐的曲线下面积大于0.7, 可以作为乙肝肝硬化伴肌肉重量减少的潜在生物标志物。结论研究筛选出33条紊乱的代谢通路和多种差异代谢物, 绝大多数紊乱的代谢通路与氨基酸代谢、尿素循环和乙醛酸盐和二羧酸盐代谢通路有关。对乙肝肝硬化患者进行差异代谢物筛选, 可以尽早识别前肌少症, 开展营养...     

Effects of Sodium Selenite Injection on Serum Metabolic Profiles in Women Diagnosed with Breast Cancer-Related Lymphedema—Secondary Analysis of a Randomized Placebo-Controlled Trial Using Global Metabolomics

In our previous study, intravenous (IV) injection of selenium alleviated breast cancer-related lymphedema (BCRL). This secondary analysis aimed to explore the metabolic effects of selenium on patients with BCRL. Serum samples of the selenium-treated (SE, n = 15) or the placebo-controlled (CTRL, n = 14) groups were analyzed by ultra-high-performance liquid chromatography with Q-Exactive Orbitrap tandem mass spectrometry (UHPLC-Q-Exactive Orbitrap/MS). The SE group showed a lower ratio of extracellular water to segmental water (ECW/SW) in the affected arm to ECW/SW in the unaffected arm (arm ECW/SW ratio) than the CTRL group. Metabolomics analysis showed a valid classification at 2-weeks and 107 differential metabolites were identified. Among them, the levels of corticosterone, LTB4-DMA, and PGE₃—which are known anti-inflammatory compounds—were elevated in the SE group. Pathway analysis demonstrated that lipid metabolism (glycerophospholipid metabolism, steroid hormone biosynthesis, or arachidonic acid metabolism), nucleotide metabolism (pyrimidine or purine metabolism), and vitamin metabolism (pantothenate and CoA biosynthesis, vitamin B₆ metabolism, ascorbate and aldarate metabolism) were altered in the SE group compared to the CTRL group. In addition, xanthurenic acid levels were negatively associated with whole blood selenium level (WBSe) and positively associated with the arm ECW/SW. In conclusion, selenium IV injection improved the arm ECW/SW ratio and altered the serum metabolic profiles in patients with BCRL, and improved the anti-inflammatory process in lipid, nucleotide and vitamin pathways, which might alleviate the symptoms of BCRL.     

Dietary Annatto-Extracted Tocotrienol Reduces Inflammation and Oxidative Stress,and Improves Macronutrient Metabolism in Obese Mice: A Metabolic Profiling Study

Obesity and its related complications are a world-wide health problem. Dietary tocotrienols (TT) have been shown to improve obesity-associated metabolic disorders, such as hypercholesterolemia, hyperglycemia, and gut dysbiosis. This study examined the hypothesis that the antioxidant capacity of TT alters metabolites of oxidative stress and improves systemic metabolism. C57BL/6J mice were fed either a high-fat diet (HFD control) or HFD supplemented with 800 mg annatto-extracted TT/kg (HFD+TT800) for 14 weeks. Sera from obese mice were examined by non-targeted metabolite analysis using UHPLC/MS. Compared to the HFD group, the HFD+TT800 group had higher levels of serum metabolites, essential amino acids (lysine and methionine), sphingomyelins, phosphatidylcholine, lysophospholipids, and vitamins (pantothenate, pyridoxamine, pyridoxal, and retinol). TT-treated mice had lowered levels of serum metabolites, dicarboxylic fatty acids, and inflammatory/oxidative stress markers (trimethylamine N-oxide, kynurenate, 12,13-DiHOME, and 13-HODE + 9-HODE) compared to the control. The results suggest that TT supplementation lowered inflammation and oxidative stress (oxidized glutathione and GSH/GSSH) and improved macronutrient metabolism (carbohydrates) in obese mice. Thus, TT actions on metabolites were beneficial in reducing obesity-associated hypercholesterolemia/hyperglycemia. The effects of a non-toxic dose of TT in mice support the potential for clinical applications in obesity and metabolic disease.     

Camellia (Camellia oleifera Abel.) Seed Oil Regulating of Metabolic Phenotype and Alleviates Dyslipidemia in High Fat-Fed Mice through Serum Branch-Chain Amino Acids

Camellia (Camellia oleifera Abel.) seed oil (CO) has been shown to effectively reduce the blood lipid level of its host due to its fatty acid content, but the specific molecular mechanism associated with the metabolic phenotype after digestion is not clear. Here, we further investigated the relationship between branched-chain amino acids (BCAA) and the metabolic phenotype that may exhibit the anti-dyslipidemia effect of CO on mice fed a high-fat diet for 30 day C57BL/6J male mice were allocated to three groups: the control group (Cont), the high-fat feed group (HFD), and a high-fat feed group with CO treatment (CO). A serum sample was collected to detect lipid biomarkers and BCAA concentration. Notably, Low-density lipoprotein (LDL), Total Cholesterol (TC), and Triglycerides (TG) showed a significant decrease, whereas High-density lipoprotein (HDL) increased in CO mice but not in the HFD group. The concentration of Isoleucine (Ile), leucine (Leu), and valine (Val) was similar between the Cont and CO groups compared with the HFD group, exhibiting an inhibition induced by CO in mice fed with a high-fat diet. A metabolic phenotype from serum examined by non-targeted metabolite analysis using UHPLC/MS showed most metabolites exhibited lipid and BCAA metabolism. The results indicated that CO treatment notably regulated the metabolism of arachidonic acid and steroid biosynthesis in response to HFD-induced dyslipidemia. In addition, the expression of PPARγ genes that correlated with the BCAA and serum lipid biomarkers were compared, and significant inhibition was noticed, which might lead to the potential exposure of the anti-dyslipidemia mechanism of CO in HFD-fed mice. In conclusion, the expression of PPARγ genes, serum lipid level, BCAA concentration, and the metabolic phenotype was significantly positive in correlation with a high-fat diet, whereas oral CO improved the biomarkers and metabolism of some specific serum metabolites in HFD-fed mice.     

Characterization of small metabolites alteration in mice brain tissues after infected by rabies virus

Rabies, caused by rabies virus (RABV), is still one of the deadliest infectious diseases. Host metabolomic changes against RABV infection has not yet been fully understood. We performed untargeted metabolomics to discover the metabolites associated with RABV infection. The brain tissues from 20 RABV infected mice and 10 mock infected mice were used for this method. A total of 1352 differential metabolites were identified after the first-run screen, and the number reduced to 75 after second-run screen. Multivariate analysis using PLS-DA and OPLS-DA clearly discriminated the RABV infected samples from controls. Pathways enrichment analysis revealed that most differential metabolites were associated with metabolism of nucleotide and amino acid, and aminoacyl - tRNA biosynthesis and purine metabolism were the most active pathways. The findings presented in our study would promote the understanding of metabolomics changes in brains of mice after RABV infection as well as a new perspective to study the relationship between RABV and host.     

不完全睡眠剥夺对大鼠甲状腺功能影响的研究

目的研究不完全睡眠剥夺对Wistar大鼠甲状腺功能及相关免疫状态的影响,并分析其可能的机制。方法健康3月龄雄性SPF级Wistar大鼠48只,随机分为睡眠剥夺18 h组(间歇6 h)、睡眠剥夺22 h组(间歇2 h)、持续光照组(24 h持续光照)、对照组(模拟正常12 h拟光照,12 h黑暗的环境),共4组,每组12只大鼠,利用"小平台水环境法"建立大鼠睡眠剥夺模型。中。21 d后检测大鼠血清三碘甲腺原氨酸(TT_3)、四碘甲腺原氨酸(TT_4)、血清游离三碘甲腺原氨酸(FT_3)、游离甲状腺素(FT_4)、促甲状腺激素(TSH)、甲状腺球蛋白抗体(Tg抗体)和甲状腺过氧化物酶抗体(TPO抗体)水平。结果睡眠剥夺组大鼠睡眠剥夺实验终末体重较起始体重下降(P0.01)、睡眠剥夺22 h组的甲状腺相对质量高于对照组(均P0.01);与对照组相比,睡眠剥夺18 h组血清TT_3、TT_4、FT_4、TSH水平上升(P0.05,P0.01);与对照组相比,睡眠剥夺22 h组血清TT_3、Tg抗体、TSH水平上升(P0.05),TT_4、FT_4水平下降(P0.01)。结论不完全睡眠剥夺使大鼠甲状腺功能受损,并影响大鼠免疫功能。