Recently, the great interests in application of natural product have led to focus on the potential impact on bio macromolecules. Propolis is one of the natural products with various biological activities such as anti-diabetic effect. Under hyperglycemia, hemoglobin (Hb) frequently undergoes non-enzymatic glycation which affect its structure and function. In this study, the binding interaction between bulk propolis and three forms of Hb, native Hb, glycated Hb (GHb) and fructated Hb (FHb), in two temperatures of 25 and 37 °C have been examined and compared with our previous study (interaction of propolis nanoparticles with Hb, GHb and FHb). The binding studied with applying fluorescence and far UV circular dichroism spectroscopic techniques. Bulk Propolis possess the ability to quench the intrinsic fluorescence of the different types Hb like propolis nanoparticles. The mechanism of interaction bulk propolis with different types Hb was similar to the propolis nanoparticles, dynamic quenching in native Hb and static quenching in glycated and fructated Hb. According to the modified Stern–Volmer equation, the binding constant and number of binding sites were obtained. The result suggested that propolis nanoparticles have more quenching rate constant (kq) in compared to the bulk propolis in interaction different types Hb. In addition, the values of binding constant (Kb) revealed that nanopropolis has more binding affinity in compared to the bulk propolis. Data obtained from thermodynamic parameters implied more portion of electrostatic forces in GHb-propolis than native Hb-propolis and more hydrophobic portion in FHb-propolis. The CD spectra of Hb, GHb and FHb displayed no significant changes in secondary structure after incubated with propolis. These data are agreement with propolis nanoparticles. Results showed that propolis nanoparticles are a stronger quencher and bound to Hb, GHb and FHb with higher affinity than bulk propolis.

近来，对天然产物应用的极大兴趣导致人们将注意力集中在对生物大分子的潜在影响上。蜂胶是具有多种生物活性如抗糖尿病作用的天然产物之一。在高血糖症下，血红蛋白（Hb）经常经历非酶糖基化，从而影响其结构和功能。在这项研究中，研究了在25和37°C的两个温度下，蜂胶与三种形式的Hb，天然Hb，糖化Hb（GHb）和果糖Hb（FHb）之间的结合相互作用，并将其与我们以前的研究进行比较蜂胶纳米粒子与Hb，GHb和FHb的相互作用）。通过应用荧光和远紫外圆二色光谱技术研究了结合。散装蜂胶具有猝灭不同类型Hb像蜂胶纳米粒子的固有荧光的能力。蜂胶与不同类型的血红蛋白相互作用的机理类似于蜂胶纳米粒子，天然血红蛋白中的动态淬灭和糖化和果糖中的血红蛋白的静态淬灭。根据改进的Stern-Volmer方程，获得了结合常数和结合位点数。结果表明，与不同类型Hb相互作用的本体蜂胶相比，蜂胶纳米粒子具有更大的猝灭速率常数（kq）。另外，结合常数（Kb）的值表明，与蜂胶相比，纳米蜂胶具有更大的结合亲和力。从热力学参数获得的数据表明，相比天然Hb蜂胶，GHb蜂胶中静电力的份额更大，而FHb蜂胶中疏水性更高的份额。与蜂胶一起孵育后，Hb，GHb和FHb的CD光谱显示二级结构无明显变化。这些数据与蜂胶纳米颗粒一致。结果表明，蜂胶纳米颗粒是更强的猝灭剂，与散装蜂胶相比，与Hb，GHb和FHb的结合力更高。