菜籽肽抗氧化活性的研究

用分光光度法测定大鼠红细胞溶血程度,用丙二醛(MDA)试剂盒测定MDA。结果显示,通过腹膜注射菜籽肽,可以使小鼠血清中MDA含量显著减少。高剂量组[100 mg/(kg.d)]与对照组相比,MDA的降低达到极显著水平(P<0.01)。离体实验结果显示,菜籽肽粗品及各级分对体外温育和H2O2诱导的小鼠肝组织匀浆MDA生成有较强的抑制作用,其中粗品RSP-R和级分RSP-1的作用最好,且有量效关系,而对Fe2 诱导的MDA生成的抑制作用稍差。另外,RSP-R还可抑制大鼠红细胞氧化溶血程度。以上结果表明,菜籽肽在体内外均具有明显的抗氧化作用。     

Factors affecting in vitro formation of tannin-protein complexes

In interaction of condensed tannins from Desmodium intortum and Lotus pedunculatus and tannic acid (hydrolysable tannin) with salivary mucoproteins (from sheep and goats), plant leaf proteins and bovine serum albumin were evaluated. These studies were carried out over a pH range of 2-0-9-0 and different inorganic ion conditions to simulate conditions in which dietary proteins would interact with tannins in a ruminant digestive tract. Insoluble tannin-protein interactions were found at pH 4–5–5–5 for bovine serum albumin and 3–5–5–5 for plant leaf protein. The present study showed that pH alone was not the sole determinant for tannin-protein complex formation, since tannin-protein complexation was found in the pH range 6-0–6-5 when different inorganic ions were added to the solutions. Insoluble complexes were not formed with salivary proteins, although precipitation by tannic acid was achieved at 5°C. This suggests that tannins may form soluble rather than insoluble complexes with salivary proteins. It was concluded that purified F1 leaf protein (the major protei occurring in leaf tissue) ought to be used as the test protein for evaluating tannin-protein interactions for in vitro assay procedures. Using this method it was calculated that 27–43% and 19–40% of available plant protein may interact with condensed tannins from Desmodium intortum and Lotus pedunculatus, respectively.     

Combined cross-linking treatments of bovine serum albumin gel beadlets for controlled-delivery of caffeine

Combined cross-linking agents (CCLA) of microbial transglutaminase (MTgase) and ribose were applied during production of bovine serum albumin gels via incubation and heating treatment, respectively. CCLA produced stronger gels with lower protein solubility in disruptive solvents (1% sodium dodecyl sulphate plus 1% β-mercaptoethanol) as compared to BSA gels (BSA/Control) or gels produced using single cross-linking agents (SCLA) of MTGase or ribose. The gels were then converted into dried beadlets containing caffeine following a freeze-drying process. In-vitro controlled-release of caffeine and swelling ratio studies of the beadlets in artificial saliva or simulated gastric fluid indicated that CCLA beadlets had the slowest release of caffeine and the lowest swelling ratio as compared to other beadlets. Scanning electron microscopy (SEM) data suggested that the improved release and the lower swelling ratio were mainly due to the denser network formed within the CCLA beadlets that had restricted the diffusion of caffeine and hampered the enzymatic breakdown of the matrix. The additional protein cross-linkings formed as a result of MTgase incubation and ribose-induced Maillard reaction could provide a delay action in releasing caffeine that potentially extend the duration of the action of the drug during ingestion.     

New metal chelate sorbent for albumin adsorption: Cibacron Blue F3GA-Zn(II) attached microporous poly(HEMA) membranes

Poly(2-hydroxyethyl methacrylate) [poly(HEMA)] membranes were prepared by UV-initiated photopolymerization of HEMA in the presence of an initiator (α-α′-azobis-isobutyronitrile, AIBN). The triazine dye Cibacron Blue F3GA was attached as an affinity ligand to poly(HEMA) membranes, covalently. These affinity membranes with a swelling ratio of 58% and containing 10.7 mmol Cibacron Blue F3GA/m² were used in the albumin adsorption studies. After dye-attachment, Zn(II) ions were chelated within the membranes via attached-dye molecules. Different amounts of Zn(II) ions [650–1440 mg Zn(II)/m²] were loaded on the membranes by changing the initial concentration of Zn(II) ions and pH. Bovine serum albumin (BSA) adsorption on these membranes from aqueous solutions containing different amounts of BSA at different pH was investigated in batch reactors. The nonspecific adsorption of BSA on the poly(HEMA) membranes was negligible. Cibacron Blue F3GA attachment significantly increased the BSA adsorption up to 92.1 mg BSA/m². Adsorption capacity was further increased when Zn(II) ions were attached (up to 144.8 mg BSA m²). More than 90% of the adsorbed BSA was desorbed in 1 h in the desorption medium containing 0.5M NaSCN at pH 8.0 and 0.025M EDTA at pH 4.9. © 1998 John Wiley & Sons, Inc. J Appl Polym Sci 68: 657–664, 1998     

Diagnosing diabetic nephropathy by 1H NMR metabonomics of serum

Object: The most severe complication of type 1 diabetes (T1DM) is diabetic nephropathy. It is associated with a high risk of cardiovascular complications and premature death and requires early detection to be efficiently treated. The clinical practice to diagnose diabetic nephropathy is also a non-optimal and tedious set up based on albumin excretion rate in multiple overnight or 24h urine samples. Conversely, in this study, these independent diagnostic data are used to provide a realistic testing case for applying ¹H NMR metabonomics of serum in a diagnostic fashion. Materials and Methods: 182 T1DM and 21 non-diabetic (non-T1DM) individuals were studied. The ¹H NMR of serum at 500 MHz was targeted at two molecular windows: lipoprotein lipids and low-molecular-weight metabolites. Results: T1DM and non-T1DM individuals were exclusively separated by ¹H NMR. For diabetic nephropathy diagnosis in the T1DM patients, ¹H NMR data (and clinical biochemistry data) gave a sensitivity of 87.1% (83.9%) and a specificity of 87.7% (95.9%). The predictive values of positive and negative tests were 89.0% (95.5%) and 83.6% (79.2%), respectively. Conclusions: ¹H NMR metabonomics clearly distinguishes metabolic characteristics of T1DM and appears approximately as good a means to diagnose diabetic nephropathy from serum as an advanced set of biochemical variables.     

吸附树脂的制备及吸附蛋白质的研究

本文用醇钠活化球状聚乙烯醇颗粒，Williamson醚化法制得了大孔径的吸附树脂。以牛血清白蛋白(BSA)为模型蛋白，研究了蛋白质在多孔吸附树脂表面的吸附行为。通过测定蛋白质的吸附等温线，并用线性最小二乘法对曲线进行拟合，发现蛋白质在疏水吸附树脂的吸附等温线可用Langmuir方程描述。     

离子型表面活性剂与牛血清白蛋白作用的比较研究

采用荧光光谱、紫外吸收光谱、动态光散射和Zeta电位法对比研究了十二烷基硫酸钠(SDS)和十二烷基三甲基溴化铵(DTAB)与牛血清白蛋白(BSA)的相互作用机理。结果表明：SDS和DTAB均能猝灭BSA内源荧光,298 K时的猝灭常数Ksv分别为2.64×104和304.21 L/mol。同步荧光光谱和三维荧光光谱显示SDS和DTAB对BSA的构象产生了影响。SDS对BSA荧光猝灭机理为动静联合猝灭机制;热力学计算表明,SDS与BSA的结合过程中,静电力起主导作用,并且能与BSA形成SDS/BSA超分子复合物;DTAB对BSA荧光猝灭机理为动态猝灭,作用力主要是疏水作用。SDS和DTAB与BSA的平均结合距离分别为2.77和4.73 nm。综合结合常数、粒径和Zeta电位等变化,在相同条件下具有较大电荷密度和较小体积极性头基的SDS与BSA具有更强的结合作用。     

Cu~(2+)、Zn~(2+)和Pb~(2+)对绿原酸与牛血清白蛋白结合作用的影响

以杜仲绿原酸(CA)为原料,在模拟人体生理条件下,采用荧光光谱和紫外吸收光谱等方法,考察Cu2+、Zn2+和Pb2+对CA与牛血清白蛋白(BSA)相互作用产生的影响。结果表明,金属离子的存在不会改变CA对BSA的猝灭类型,但对其猝灭速率常数(kq)、结合常数(K)以及结合位点(n)会有不同程度的影响;CA与BSA之间的反应属于自发型,主要以范德华力和氢键为主,Zn2+和Pb2+的存在不会改变其作用力类型,Cu2+则会使作用力变为以静电吸引力为主;Zn2+和Pb2+会将供体与受体之间的结合距离分别减小1.975%和14.07%,Cu2+的干扰使CA与BSA之间的结合距离增大11.58%;同步荧光光谱显示CA与BSA中的酪氨酸残基和色氨酸残基都有相互作用,三维荧光测定结果进一步表明猝灭剂对BSA构象及微环境会引起变化。