The changes of lysozyme conformation in the absence and presence of luteolin and luteoloside were investigated by spectral analysis including fluorescence, UV, CD, Raman, and ATR-FTIR, and the biological activity of lysozyme was investigated by lysozyme assay kit. The results showed that the microenvironment hydrophobicity of lysozyme increased and peptide extension decreased with the addition of luteolin or luteoloside. The α-helix of lysozyme might be influenced by luteolin or luteoloside, and its relative content had a significant difference after adding luteolin or luteoloside by the ATR-FTIR method, which was reconfirmed by CD and Raman spectra. The lysozyme activity changed obviously after adding luteolin or luteoloside. All of the conclusions above indicated the active site of lysozyme in the α-helix might be influenced by luteolin and luteoloside.

The interactions of Salvianolic acis A (SAA) and Salvianolic acid B (SAB) with insulin were studied by using fluorescence spectroscopy, UV-vis spectroscopy and ATR-FTIR spectroscopy in simulating physiological condition (pH 7.40). The fluorescence quenching of insulin by SAA and SAB were static quenching process. The results of synchronous fluorescence and three-dimensional fluorescence spectra suggested no obvious conformation changes of insulin after SAA or SAB binding. But ATR-FTIR spectra showed that SAA and SAB could change the secondary structures of insulin, of which β-turns decreased and random coil increased accompanied with α-belices and β-sheets no clear change. The glucose might influenced the bioactivity of insulin in the SAA-insulin and SAB-insulin systems by changing the binding constants of SAA (or SAB) with insulin and exacerbating the changes of insulin conformation and relative contents of α-belices.

探讨中药补肾固筋方对SD大鼠膝骨关节炎中IL-1、TNF-α的抑制作用.44只SD大鼠随机分为正常组、予生理盐水的模型组、美洛昔康组和补肾固筋方组,除正常组外均采用右后膝改良Hulth法造模,连续干预12周,取血清及股骨内髁关节软骨、滑液及软骨下骨,ELISA法检测IL-1(白细胞介素-1)、iNOS(诱导型一氧化氮合酶)含量.结果发现美洛昔康组关节间隙狭窄、关节面粗糙,骨赘明显重于补肾固筋方组;补肾固筋方组关节间隙稍窄伴轻微骨赘.模型组IL-1、iNOS含量较正常组升高(P＜0.01);补肾固筋方组和美洛昔康组含量比模型组降低(P＜0.01)且两组之间差异无统计学意义(P＞0.05).提示补肾固筋方通过有效抑制IL-1、iNOS的分泌促进骨关节炎软骨修复.

The interaction of rutin with lysozyme(LYZ) was studied in simulating physiological condition(pH 7.40) at 298 K, 304 K and 310 K by ultraviolet-vis(UV) absorption and fluorescence spectroscopy. The results showed that rutin quenched the endogenous fluorescence of LYZ via a static quenching procedure and the interaction between them was a spontaneous process. The number of binding sites was approximately 1. Van der Waals' forces and hydrogen bonds played major roles in stabilizing rutin-LYZ complex, and the distance between the donor and acceptor was 4.02 nm(298 K). The UV absorption, synchronous fluorescence and three-dimensional fluorescence spectra showed that the conformation of LYZ became more tightly packed. The turbidimetric analysis showed that rutin decreased LYZ activity. Rutin may be bad for the hydrogen-bonding receptor catalysis activity of Asp-52 by affecting the micro-environment of lysozyme activity site(Asp-52).

Angelica sinensis (Oliv.) Diels combined treatment with warfarin would increase the risk of bleeding. Ferulic acid is an abundant hydroxycinnamic acid in A. sinensis and warfarin is the most widely used oral anticoagulant. The studies on supermolecular interaction of warfarin with human serum albumin (HSA) and the influence of ferulic acid on the binding would contribute to the understanding of the metabolic processes of warfarin and the effect of ferulic acid. We focus on investigating the effect of warfarin on fluorescence spectrum of human serum albumin (HSA), fluorescence quenching mechanism, binding constant, Hill coefficient, binding mode, and the effect of different ferulic acid concentrations on the binding. Warfarin quenched the intrinsic fluorescence of HSA mainly by static quenching. Accession of ferulic acid reduced the binding of HSA-warfarin. By decreasing binding constant and the Hill coefficient of warfarin with HSA, ferulic acid could improve the plasma concentration of free warfarin, which would increase the risk of bleeding. Warfarin’s free concentration increased at least 50% under the condition of simulated human body. The results indicated that A. sinensis combined treatment with warfarin would increase the risk of bleeding. And the results provide an important theoretical support for warfarin used as oral anticoagulant.