小麦肽对受环磷酰胺免疫抑制小鼠的免疫调节及抗氧化功能

本研究旨在分析小麦蛋白活性肽对免疫抑制小鼠免疫功能和抗氧化功能的调节作用。小鼠灌胃小麦肽10d,第8天用环磷酰胺诱导免疫抑制,测定血清溶血素、抗体生成细胞含量、脾细胞增殖、体外腹腔巨噬细胞吞噬能力、肝脏抗氧化酶活性和丙二醛(MDA)含量以及血清清除DPPH和·OH的能力。实验结果表明,环磷酰胺处理显著的降低了小鼠血清中抗SRBC抗体(溶血素HC50)水平和腹腔巨噬细胞的吞噬能力;同时伴随着肝脏超氧化物歧化酶活性(SOD)、过氧化氢酶活力(CAT)、总抗氧化能力(T-AOC)的降低和MDA含量的提高。给小鼠灌胃小麦肽可以恢复HC50和脾细胞增殖,显著提高抗体生成细胞含量和腹腔巨噬细胞吞噬能力;此外,小麦肽增强了小鼠血清清除DPPH和清除·OH的能力。以上结果表明,小麦肽可以调节应激状态引起的机体抗氧化体系紊乱及免疫功能的降低。这可能与小麦肽缓冲自由基生成、激活腹腔巨噬细胞和脾淋巴细胞活性有关。     

食源性抗氧化肽研究进展

生物活性肽主要水解自食源性蛋白质,此类活性物质表现出多种生物功能,其中尤以抗氧化能力最为突出。多项研究已证实抗氧化成分的摄入与疾病的发生呈逆相关关系。生物活性肽的抗氧化作用主要表现在自由基的清除能力、脂质过氧化的抑制能力以及金属离子的螯合能力。抗氧化活性的强弱取决于活性肽的结构及其特异性氨基酸序列。本文在对氧化应激的发生进行了介绍后,先后阐述了抗氧化肽的益生功能、酶法制备工艺、抗氧化活性的检测方法、生物利用度及其食用安全性。     

小麦低聚肽的功能作用研究进展及应用前景展望

综述小麦低聚肽抗氧化、调节免疫功能、降血压等功能作用的研究进展和应用展望,为小麦低聚肽的进一步开发利用提供思路。     

马鹿茸血免疫活性肽的制备及其活性研究

本文以免疫活性和DPPH·的清除能力为指标,研究了用木瓜蛋白酶与中性蛋白酶水解马鹿茸血制备活性肽的条件,并初步探讨了此活性肽的免疫活性与对DPPH·清除能力之间的关系.实验结果表明:当[E/S]为9000 U/g时,中性蛋白酶和木瓜蛋白酶的水解温度各为50和45 ℃,pH各为7.1和6.8,分别水解2和1.5 h可以获得最佳的酶解效果.在相同蛋白含量的情况下,两酶复合水解物的淋巴细胞增殖率比单酶水解增加了20.8%.免疫活性与DPPH·清除能力之间存在一定的相关性(r=0.957,P<0.01).     

海藻肽的化学结构特征和活性作用研究进展

海藻中的肽类化合物具有显著的生物活性和药理作用,对其氨基酸序列及活性作用的研究已经取得了一些重要进展。发现的海藻肽类化合物并确定其化学结构式的主要有二肽、环肽和脂肽,这些肽类化合物具有抗肿瘤、降血压、降血脂、抗凝血、促进神经细胞分化、抗氧化、抗菌和抗病毒等生物活性。预测海藻肽类化合物在疑难病症的治疗上将发挥重要作用。     

猪皮胶原蛋白水解产物中的抗氧化活性肽的分离及其氨基酸组分

利用不同蛋白酶水解猪皮胶原蛋白,根据水解产物对自由基的清除率,筛选最适蛋白酶的组合,筛选抗氧化活性较高的肽类,并进行氨基酸分析.结果表明:胃蛋白酶→木瓜蛋白酶→菠萝蛋白酶为酶解胶原蛋白的最佳组合;分离纯化得到了三种具有较高抗氧化活性的肽;氨基酸组分分析显示:P1′、P2′和P3′组分具有自由基清除作用的关键氨基酸甲硫氨酸、酪氨酸和组氨酸等.     

海蜇胶原蛋白肽的降血脂及抗氧化作用的研究

本研究以海蜇胶原蛋白为原料,经酶解得到海蜇胶原蛋白肽,探讨海蜇胶原蛋白肽对小鼠血脂的影响及抗氧化作用。以高脂饲料喂养ICR小鼠,建立高脂血症模型,研究胶原蛋白肽对小鼠肝系数和脂肪系数的变化情况及小鼠血脂水平、肝组织抗氧化功能的影响。结果表明海蜇胶原蛋白肽能显著降低小鼠肝系数和脂肪系数,能显著降低高脂血症小鼠血清总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、动脉硬化指数(LDL-C/HDL-C)水平,升高高密度脂蛋白胆固醇(HDL-C)和抗动脉粥样硬化因子(HDL-C/TC);能提高肝组织超氧化物歧化酶(SOD)谷胱甘肽过氧化物酶(GSH-Px)的活力,并能减低丙二醛(MDA)的含量。海蜇胶原蛋白肽具有辅助减低血脂水平和增强抗氧化功能的作用。     

绣球菌多糖表征及其抗氧化和免疫活性

提取纯化绣球菌多糖(Sparassis latifolia polysaccharides,SCPs),研究其表征和功能活性,探索绣球菌多糖表征与其抗氧化及免疫活性之间的关系。以绣球菌子实体为原料,采用聚能超声波辅助水提醇沉法提取绣球菌多糖,经DEAE-52、SephadexG-100纯化,用高效凝胶渗透色谱法、离子色谱法、傅里叶红外色谱法、扫描电镜、原子力显微镜对绣球菌多糖进行初步表征,检测绣球菌多糖清除DPPH、·OH、O2^-·自由基能力以及总还原力,用MTT法检测绣球菌多糖对巨噬细胞RAW264.7增殖的影响。结果表明,SCPs分子量范围为215Da–393kDa,由葡萄糖、甘露糖、半乳糖、木糖、果糖构成,摩尔比13:4:1:2:3,其表观形貌为簇状堆积,交织,结构规律性不强,表面光滑,呈一定的网络状结构,分子呈现链状构象,具有高度的分支结构,链间形成小环且伴随一定的球形颗粒。SCPs具有一定的还原能力和清除DPPH、·OH、O2^-·自由基的能力,且能够促进巨噬细胞RAW264.7的增殖。绣球菌多糖的抗氧化及免疫活性可能与其分子量、单糖组成、糖链分支及分子构象有关。     

小麦胚芽活性肽通过AMPK/SIRT1改善大鼠肝脏衰老损伤的作用机制研究

小麦胚芽作为小麦面粉加工过程中产生的副产品,资源极为丰富,但一直以来未能得到充分合理的利用.本研究以课题组前期获得的麦胚活性肽ADWGGPLPH(ADW)为对象,以自然增龄的SD大鼠作为肝脏老化损伤的动物模型, 9月龄大鼠作为对照,用麦胚源活性肽持续灌胃干预至21月龄,研究麦胚源活性肽对自然增龄大鼠肝脏老化损伤的影响,并结合H₂O₂诱导衰老HepG2细胞模型初步探明其分子机制.结果表明,麦胚源活性肽ADW能够增加大鼠血清及肝脏中SOD, T-AOC, GSH-Px, CAT等抗氧化酶活性,同时降低MDA水平,改善大鼠肝脏中氧化应激水平;降低血清及肝脏中IL-6, IL-1β, MMP3, TNF-α水平,并且增加抗炎因子IL-10分泌,改善增龄大鼠肝脏中炎症环境;麦胚源活性肽干预后肝脏结构明显好转、脂肪空泡减少、细胞排列整齐, ALT以AST水平均显著下降.麦胚源活性肽能够有效地改善老化肝脏中细胞的结构及生长状态异常,减少增龄过程中肝脏质量损失,降低了衰老标志蛋白P53和P21的表达量,减轻肝脏老化损伤程度并减缓了肝脏的老化进程.通过加入抑制...     

黑豆多肽分离及其抗氧化活性的研究

采用凝胶柱层析法分离具有抗氧化活性的黑豆多肽,并分析其分子量分布;同时还采用DPPH·清除法研究各分离肽段的抗氧化活性,对抗氧化活性最佳的肽段进行动力学分析.结果表明:黑豆多肽经分离后得到3个片段,黑豆小分子肽BSP3对DPPH·自由基具有最强清除作用,其半清除浓度(HSC50)为0.664 mg/mL.所以,分子量小于1000 Da的黑豆小分子肽具有很好的抗氧化活性.     

Reactive Center Loop (RCL) Peptides Derived from Serpins Display Independent Coagulation and Immune Modulating Activities

Serpins regulate coagulation and inflammation, binding serine proteases in suicide-inhibitory complexes. Target proteases cleave the serpin reactive center loop scissile P1–P1′ bond, resulting in serpin-protease suicide-inhibitory complexes. This inhibition requires a near full-length serpin sequence. Myxomavirus Serp-1 inhibits thrombolytic and thrombotic proteases, whereas mammalian neuroserpin (NSP) inhibits only thrombolytic proteases. Both serpins markedly reduce arterial inflammation and plaque in rodent models after single dose infusion. In contrast, Serp-1 but not NSP improves survival in a lethal murine gammaherpesvirus68 (MHV68) infection in interferon γ-receptor-deficient mice (IFNγR^−/−). Serp-1 has also been successfully tested in a Phase 2a clinical trial. We postulated that proteolytic cleavage of the reactive center loop produces active peptide derivatives with expanded function. Eight peptides encompassing predicted protease cleavage sites for Serp-1 and NSP were synthesized and tested for inhibitory function in vitro and in vivo. In engrafted aorta, selected peptides containing Arg or Arg-Asn, not Arg-Met, with a 0 or +1 charge, significantly reduced plaque. Conversely, S-6 a hydrophobic peptide of NSP, lacking Arg or Arg-Asn with −4 charge, induced early thrombosis and mortality. S-1 and S-6 also significantly reduced CD11b⁺ monocyte counts in mouse splenocytes. S-1 peptide had increased efficacy in plasminogen activator inhibitor-1 serpin-deficient transplants. Plaque reduction correlated with mononuclear cell activation. In a separate study, Serp-1 peptide S-7 improved survival in the MHV68 vasculitis model, whereas an inverse S-7 peptide was inactive. Reactive center peptides derived from Serp-1 and NSP with suitable charge and hydrophobicity have the potential to extend immunomodulatory functions of serpins.     

纳豆中抗氧化肽的分离纯化与活性研究

从发酵的纳豆中提取具有抗氧化活性的多肽,通过分子筛层析和反相高效液相色谱对纳豆上清液进行分离纯化,并采用电喷雾串联质谱进行结构鉴定.结果表明:纳豆发酵后的蛋白(多肽)混合物经Sephadex G-50凝胶色谱进行分离纯化后得到3个组分(F1、F2和F3),其中组分F3的抗氧化活性最强,总还原力达到(8.4±0.6)mm...     

Modulating Uranium Binding Affinity in Engineered Calmodulin EF-Hand Peptides: Effect of Phosphorylation

To improve our understanding of uranium toxicity, the determinants of uranyl affinity in proteins must be better characterized. In this work, we analyzed the contribution of a phosphoryl group on uranium binding affinity in a protein binding site, using the site 1 EF-hand motif of calmodulin. The recombinant domain 1 of calmodulin from A. thaliana was engineered to impair metal binding at site 2 and was used as a structured template. Threonine at position 9 of the loop was phosphorylated in vitro, using the recombinant catalytic subunit of protein kinase CK2. Hence, the T₉TKE₁₂ sequence was substituted by the CK2 recognition sequence TAAE. A tyrosine was introduced at position 7, so that uranyl and calcium binding affinities could be determined by following tyrosine fluorescence. Phosphorylation was characterized by ESI-MS spectrometry, and the phosphorylated peptide was purified to homogeneity using ion-exchange chromatography. The binding constants for uranyl were determined by competition experiments with iminodiacetate. At pH 6, phosphorylation increased the affinity for uranyl by a factor of ∼5, from K_d = 25±6 nM to K_d = 5±1 nM. The phosphorylated peptide exhibited a much larger affinity at pH 7, with a dissociation constant in the subnanomolar range (K_d = 0.25±0.06 nM). FTIR analyses showed that the phosphothreonine side chain is partly protonated at pH 6, while it is fully deprotonated at pH 7. Moreover, formation of the uranyl-peptide complex at pH 7 resulted in significant frequency shifts of the ν_as(P-O) and ν_s(P-O) IR modes of phosphothreonine, supporting its direct interaction with uranyl. Accordingly, a bathochromic shift in ν_as(UO₂)²⁺ vibration (from 923 cm⁻¹ to 908 cm⁻¹) was observed upon uranyl coordination to the phosphorylated peptide. Together, our data demonstrate that the phosphoryl group plays a determining role in uranyl binding affinity to proteins at physiological pH.     

功能性抗氧化肽制备与机制研究进展

抗氧化肽作为食品和机体的抗氧化剂具有广阔应用前景,通过酶水解蛋白质能得到天然抗氧化肽.本文对水解蛋白来源抗氧化肽的制备和抗氧化机制进行了综述.     

微生物源性抗氧化剂对小鼠抗氧化性能及免疫功能的影响

旨在探究不同剂量的微生物源性抗氧化剂对小鼠抗氧化性能及免疫功能的影响。将60只28日龄雌性昆明小鼠随机分为4组,每组15只。试验组分别以0.5 g/kg bw*d(低)、1.0 g/kg bw*d(中)、1.5 g/kg bw*d(高)的剂量灌喂微生物源性抗氧化剂,每天1次,连续30 d,灌胃容量为0.2 mL/10 g bw*d,对照组灌喂等容量的蒸馏水。30 d后,测定微生物源性抗氧化剂对小鼠抗氧化性能及免疫功能的影响。结果表明,1.0 g/kg bw*d的微生物源性抗氧化剂可以极显著地提高血清中GSH-Px的活力(P<0.01),显著提高T-SOD活力(P<0.05),降低MDA的含量(P>0.05)。而1.5g/kg bw*d的微生物源性抗氧化剂可以极显著地提高血清中IgA的含量(P<0.01)和IL-2水平,以及ConA刺激的脾淋巴细胞转化率(P<0.01),促进脾脏和胸腺的发育(P>0.05)。提示1.0g/kg bw*d的微生物源性抗氧化剂可以显著增强小鼠的抗氧化能力,1.5 g/kg bw*d的微生物源性抗氧化剂可以增强免疫功能。     

Effect of Heavy Metals on Wheat Seedlings: Activation of Antioxidant Enzymes

Accumulation of heavy metals in wheat grain exposed to multicomponent pollutants (industrial wastewater) was studied. The absolute content of metals (Zn, Cd, Cu, Cr, Ni, Co, Pb, and Mn) was found to be determined by the extent of purification of wastewater. An increase in the degree of grain contamination with heavy metals was accompanied by activation of antioxidant enzymes (superoxide dismutase, EC 1.15.1.1; catalase, EC 1.11.1.6; and peroxidase, EC 1.11.1.7) in leaves and activation of superoxide dismutase and peroxidase in roots. The ratio of activity of membrane enzymes to activity of cytosol enzymes was demonstrated to be high. It was concluded that the membranotropic effect of multicomponent contaminants was due to accumulation of heavy metals capable of inducing the antioxidant protection in the next generation of wheat seedlings.     

双酶法酶解豌豆蛋白制备高抗氧化多肽的研究

以水解度及DPPH自由基清除能力为指标优选出1398中性蛋白酶和胰蛋白酶分步酶解豌豆分离蛋白,所得豌豆蛋白水解产物(pea protein hydrolysate,PPH)中相对分子量小于2 KDa的多肽占77.07％,对DPPH的半清除浓度(IC50)为3.01 mg/mL;PPH经Sephadex G-25凝胶层析和两次RESOURCETM 3RPC反相层析纯化获得高抗氧化活性峰,经反相层析鉴定为层析纯,当浓度在0.2 mg/mL时,对DPPH的清除率达到62.03％,与同浓度下的谷胱甘肽对DPPH的清除率相近(66.82％).     

Antioxidant Synergetic Effect Between the Peptides Derived from the Egg White Pentapeptide Trp-Asn-Trp-Ala-Asp

When foods are intaken together, the antioxidant activity of mixtures may present synergistic, additive, or antagonistic interactions. The aim of this study was to investigate the oxygen radical absorbance capacity (ORAC) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonicacid) diammonium salt (ABTS) radical scavenging activity of (Trp-Asn-Trp-Ala-Asp) WNWAD, a pentapeptide derived from egg white ovomucin pepsin hydrolysates, and its fragments as well as the effect of WNWAD on cellular antioxidant enzymes activities. The ORAC assays demonstrated that combined group (WN?+?WAD) significantly improved the antioxidant activities, while combined group (WNW?+?AD) decreased the antioxidant activities. The ABTS assays showed peptide fragments (WNW, WN, WAD) and its mixture (WNW?+?AD, WN?+?WAD) have lower EC₅₀ than antioxidant peptide WNWAD in ABTS assay. Various combinations of peptide were found to have synergistic interaction, and synergistic interaction exhibited certain dose-dependently interactions. At cellular level, the activities of SOD, GSH-Px, CAT were higher while the level of MDA, LDH release were lower in the peroxide?+?peptide-treated group when compared to the peroxide exposed group. Results in this study imply the importance of exploiting synergism interactions in the development of new functional food.