消化过程被视为食物与健康之间的生理桥梁，其中组分间相互作用会影响营养素的生物可及性。这一关键过程决定了人体对营养成分的吸收效率，从而直接影响身体的营养状况和健康水平。蛋白质和脂质作为食物中的两大主要营养成分，在人体代谢中发挥着重要作用。然而，高脂饮食下蛋白质与脂质的交互作用如何调节营养组分的释放速度和生物利用度尚不明确。本研究使用体外消化模型，聚焦于揭示脂质和蛋白在消化过程中的交互影响，以确定膳食蛋白种类及脂肪含量是否会影响脂质和蛋白的消化速率。重点关注蛋白质与生理表面活性剂胆盐之间的相互作用，以及它们对脂质消化的调节功能。本研究旨在进一步阐明蛋白质和脂质在食物消化过程中的协同作用机制，具体的研究如下：

1.蛋白质种类和脂肪含量对膳食结构及消化特性的影响研究

本章旨在探明全营养膳食中蛋白与脂肪的交互作用及其对营养素生物利用度的影响。选择四种不同来源的蛋白质（酪蛋白、猪肉蛋白、鸡肉蛋白和大豆蛋白）并通过添加全营养素制备了标准膳食和高脂膳食。在体外消化模型下测定了不同膳食组分间的微观结构变化及蛋白和脂质的水解速率，以期确定蛋白和脂质在膳食体系中的相互作用。结果表明，脂肪含量的增加会影响蛋白质的二级和三级结构，蛋白质疏水基团暴露，溶解度降低。高脂膳食中蛋白与脂肪具有更高的乳化活性，进而提升了猪肉蛋白和鸡肉蛋白的消化率。不同的蛋白质与脂质的微观结合状态存在差异，并最终导致体系中脂质水解速率不同，具体表现为肉蛋白膳食中的脂质水解速率低于酪蛋白和大豆蛋白膳食。这一发现可以为了解高脂膳食中蛋白质与脂质的协同消化作用提供新的见解。

2.高脂膳食条件下不同蛋白质对脂质水解的影响及内在机制

蛋白种类和高脂肪水平是影响营养素利用的重要因素，然而其内在机制尚不明确。本章研究了不同蛋白质来源（猪肉蛋白、鸡肉蛋白、酪蛋白和大豆蛋白）的高脂高蛋白膳食的体外消化行为，以期探究蛋白质种类对脂质水解的影响机制。尽管脂肪类型和水平相同，但不同蛋白膳食中脂质最终水解程度存在差异，酪蛋白膳食中脂质水解度最高，而猪肉和鸡肉蛋白中的脂质水解度较低。这种差异归因于猪肉和鸡肉中的盐溶性蛋白（肌原纤维蛋白）在小肠消化过程中能有效结合胆盐，降低了其参与脂肪消化的效率。平衡吸附实验和荧光光谱结果表明，猪肉和鸡肉的肌原纤维蛋白对胆盐具有较强的结合能力，疏水作用在结合程度中起着至关重要的作用。此外，不同蛋白质来源膳食的流变特性和微观结构也有所不同。肉类蛋白质膳食表现出较强的聚集作用，导致反应的界面面积较小。本章通过探讨不同膳食蛋白在消化道中的聚集性差异以及蛋白质与胆盐的相互作用，初步解析了蛋白质对脂质水解的影响机制。

3. 蛋白与胆盐相互作用机制及其在脂质界面层吸附动力学研究

本章研究旨在进一步探究小肠消化阶段蛋白质消化物与胆盐的相互作用以及它们在脂质界面层上的吸附动力学规律。通过结合等温实验和圆二色谱分析，发现肉源性肌原纤维蛋白可以结合更多的胆盐分子，胆盐的加入会增加蛋白质二级结构的展开过程。荧光光谱和粒径分析确定了牛磺脱氧胆酸钠与蛋白之间具有最强的相互作用，胆盐的加入会阻止蛋白质的自缔合并降低聚集性。不同蛋白在脂质层上的最终界面压力有所差异，酪蛋白最高（14.77 ± 0.12 mM/m），猪肉和鸡肉肌原纤维蛋白最低，分别为12.08 ± 0.25和10.75 ± 0.35 mM/m。蛋白质-胆盐混合物在界面上表现出比单一胆盐更高的界面压力和渗透速率，胆盐与蛋白质之间的相互作用使得胆盐无法完全置换蛋白质，这可能成为脂肪酶水解脂质的限速步骤。本章从蛋白-胆盐互作及界面张力变化两个方面进一步解释了肉源性蛋白对脂质水解的抑制机理。

4. 蛋白与脂质的体相相互作用及其对脂肪酶扩散速率的影响研究

本章旨在探究蛋白质-脂质体系在消化过程中的动态行为，包括体相相互作用及对脂肪酶扩散速率的影响。利用扩散波谱光谱微流变仪检测了不同蛋白稳定的乳液的粘弹性特征，并采用多重光散射监测了消化前后乳液的稳定性。研究结果表明，肌原纤维蛋白稳定的乳液相对于其他蛋白具有更高的粘弹性。通过微观表征和电位观察了不同蛋白乳液在小肠消化过程中的动态行为。发现不同蛋白的界面膜厚度存在差异，其中鸡肉肌原蛋白的界面膜最厚，为2.92 ± 1.24 μm，而酪蛋白的界面膜厚度仅为1.54 ± 0.47 μm。蛋白的存在造成胆盐临界胶束的延迟，其中猪肉和鸡肉肌原纤维蛋白作用更为明显。通过荧光漂白后恢复技术，FITC-胰脂肪酶在猪肉肌原纤维蛋白和鸡肉肌原纤维蛋白乳液中的扩散系数低于其他组别。肌原纤维蛋白抑制脂质水解的机制进一步被解释。

综上所述，本研究探讨了蛋白质与脂质在体外消化模型中的相互作用，重点关注高脂膳食水平下不同蛋白质对脂质水解速率的调控机制。相较于酪蛋白和大豆蛋白膳食，肉蛋白膳食中脂质具有较低的水解速率。这主要是因为肉蛋白膳食在消化道环境中更聚集，与胆盐有强烈的相互作用，从而降低了脂肪酶的水解效率。肌原纤维蛋白由于其强疏水性和特殊的壁垒效应，被确定为肉蛋白中抑制脂质水解的主要功能成分。

The digestive process is considered a physiological bridge between food and health, where interactions between components affect nutrient bioavailability. This critical process determines the efficiency of nutrient absorption by the human body, thereby directly impacting nutritional status and health levels. Proteins and lipids, as two major nutrients in food, play important roles in human metabolism. However, the mechanisms by which the interaction between proteins and lipids under high-fat dietary conditions regulates the release rate and bioavailability of nutrients remain unclear. This study uses an in vitro digestion model to elucidate the interactive effects of lipids and proteins during digestion, focusing on how different dietary proteins and fat content influence the digestion rates of lipids and proteins. Special emphasis is placed on the interactions between proteins and physiological surfactants, such as bile salts, and their regulatory functions in lipid digestion. The study aims to further elucidate the synergistic mechanisms of proteins and lipids during the digestion process, with the specific research outlined as follows:

1. Effects of Protein Types and Fat Content on Dietary Structure and Digestive Characteristics

The aim of this chapter is to elucidate the interaction between proteins and lipids in a complete nutritional diet and its impact on nutrient bioavailability. Four different sources of proteins (casein, pork protein, chicken protein, and soy protein) were selected, and standard diets and high-fat diets were prepared by adding all nutrients. Microstructural changes between different dietary components and the efficiency of protein and lipid hydrolysis were measured using an in vitro digestion model to determine the interaction between proteins and fats in the dietary system. The results indicated that increasing fat content affects the secondary and tertiary structure of proteins, exposing hydrophobic groups and reducing solubility. Proteins and lipids in high-fat diets exhibited higher emulsification activity, thereby enhancing the digestibility of pork protein and chicken protein. Different proteins exhibited distinct microstructural binding states with lipids, resulting in different rates of lipid hydrolysis in the system, with the lipid hydrolysis rate in meat protein diets being lower than that in casein and soy protein diets. This finding provided new insights into the synergistic digestion of proteins and lipids in high-fat diets.

2. The Effects and Underlying Mechanisms of Different Proteins on Lipid Hydrolysis under High-Fat Diet Conditions

Protein type and high-fat levels are important factors influencing nutrient utilization, however, the underlying mechanisms remain unclear.  This chapter investigated the in vitro digestion behavior of high-fat, high-protein diets sourced from different proteins (pork protein, chicken protein, casein, and soy protein) to explore the mechanisms by which protein types affect lipid hydrolysis. Despite the same type and level of lipid, differences in the final degree of lipid hydrolysis were observed among different protein diets. Casein-based diets exhibited the highest lipid hydrolysis, while lipid hydrolysis in pork and chicken protein diets was comparatively lower. This disparity was attributed to the effective binding of salt-soluble proteins (myofibrillar proteins) in pork and chicken during small intestinal digestion, which reduced their efficiency in lipid digestion. Equilibrium adsorption experiments and fluorescence spectroscopy results indicated that myofibrillar proteins from pork and chicken had strong binding abilities with bile salts, with hydrophobic interactions playing a crucial role in the degree of binding. Furthermore, diets from different protein sources exhibited distinct rheological properties and microstructures. Meat protein diets demonstrated strong aggregation, resulting in a smaller interfacial area for reactions. The findings of this chapter provided valuable insights into the potential mechanisms by which diets rich in different protein sources modulate fat digestion.

3. Protein-bile Salt Interaction Mechanism and its Adsorption Kinetics at the Lipid Interface

This chapter aimed to further investigate the interaction between protein digestion products and bile salts during the small intestinal digestion phase, as well as their adsorption kinetics at the lipid interface. Through combined isothermal experiments and circular dichroism analysis, it was found that meat-derived myofibrillar proteins could bind more bile salt molecules, with the addition of bile salts enhancing the unfolding process of protein secondary structure. Fluorescence spectroscopy and particle size analysis confirmed that sodium taurodeoxycholate exhibited the strongest interaction with proteins, with the addition of bile salts preventing protein self-aggregation and reducing aggregation. Different proteins exhibited varying final interfacial pressures on the lipid layer, with casein being the highest (14.77 ± 0.12 mM/m), and pork and chicken myofibrillar proteins being the lowest, at 12.08 ± 0.25 and 10.75 ± 0.35 mM/m, respectively. Protein-bile salt mixtures showed higher interfacial pressure and permeation rate at the interface compared to single bile salts, as the interaction between bile salts and proteins prevented complete displacement of proteins, which may have become a rate-limiting step in lipase-mediated lipid hydrolysis. This chapter further explained the inhibitory mechanism of meat-derived proteins on lipid hydrolysis from the perspectives of protein-bile salt interactions and changes in interfacial tension.

4. Study on the Phase Interactions between Protein and Lipid and their Effects on the Diffusion Rate of Lipase

This chapter aimed to investigate the dynamic behavior of protein-lipid systems during digestion, including their phase interactions and their impact on the diffusion rate of lipases. Using diffusion spectroscopy and spectroscopic micro-rheology, the viscoelastic properties of emulsions stabilized by different proteins were examined, and the stability of emulsions before and after digestion was monitored using multiple light scattering. The results showed that emulsions stabilized by myofibrillar proteins exhibited higher viscoelasticity compared to other proteins. The dynamic behavior of different protein emulsions during intestinal digestion was characterized through microscopic observation and zeta potential measurement. It was found that the thickness of interfacial membranes varied among different proteins, with chicken myofibrillar protein having the thickest membrane at 2.92 ± 1.24 μm, while casein had a membrane thickness of only 1.54 ± 0.47 μm. The presence of proteins delayed the formation of the critical micelle concentration of bile salts, with pork and chicken myofibrillar proteins showing more pronounced effects. Using the fluorescence recovery after photobleaching (FRAP) technique, it was observed that the diffusion coefficient of FITC-labeled pancreatic lipase in pork myofibrillar protein and chicken myofibrillar protein emulsions was lower compared to other groups. The mechanism of inhibition of lipid hydrolysis by myofibrillar proteins was further elucidated.

In summary, this study investigated the interactions between proteins and lipids in an in vitro digestion model, with a focus on the regulatory mechanisms by different proteins influence the rate of lipid hydrolysis under high-fat dietary conditions. Compared to diets containing casein and soy protein, diets with meat protein exhibited lower lipid hydrolysis rates. This is mainly attributed to the meat protein diet being more aggregated in the digestive tract environment and having strong interactions with bile salts, thereby reducing the efficiency of lipid hydrolysis by lipases. Myofibrillar proteins, due to their strong hydrophobicity and specific barrier effects, have been identified as the primary functional components inhibiting lipid hydrolysis in meat protein.

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