功能性人工油体制备工艺优化及网络药理学预测其干预肥胖的作用靶点

Preparation process optimization of functional artificial oil bodies and prediction of their obesity-alleviating targets via network pharmacology

摘要:

人工油体(artificial oil bodies, AOB)凭借仿生天然油体的结构优势与生物相容性，在活性成分靶向递送领域展现出独特的应用潜力。针对传统人工油体功能单一及体系稳定性不足的问题，本研究通过优化制备工艺，构建了高效稳定的功能性人工油体递送体系。采用单因素实验结合Box-Behnken响应面法，系统优化AOB制备工艺；通过分子对接手段预测茶多酚(tea polyphenols, TP)与AOB结构蛋白Ctcaleosin的结合能力，进一步采用超声自由基法将TP与AOB进行修饰偶联，构建茶多酚-蛋白复合物介导的功能性人工油体(AOB@TP)；基于网络药理学技术系统预测AOB@TP中茶多酚与中链甘油三酯(medium-chain triglycerides, MCT)协同干预肥胖的作用靶点。优化所得AOB制备工艺参数为：MCT 10 mg、蛋黄卵磷脂100 μg、Ctcaleosin蛋白275 μg；在此条件下，茶多酚与AOB的结合率高达94.55%，AOB@TP呈现类球形结构，粒径为216.67 nm，具备优良的载体特性；网络药理学预测结果显示，茶多酚与MCT共筛选出潜在作用靶点67个，与肥胖治疗相关的交集靶点60个，其中MMP9、HIF1A为核心调控靶点；GO功能富集分析表明其主要参与脂质代谢、能量稳态等生物学过程，KEGG通路富集主要涉及肥胖相关信号通路。本研究构建的AOB@TP功能性递送体系，兼具粒径小、多功能等优势，不仅为脂溶性营养物质的高效递送提供了新型优质载体，也为功能性人工油体在代谢疾病干预中的深入应用奠定了理论基础与技术基础。

Abstract:

Artificial oil bodies (AOBs), leveraging the structural advantages of biomimetic natural oil bodies and their inherent biocompatibility, demonstrate unique potential in the field of targeted delivery of bioactive compounds. To address the issues of single functionality and inadequate systemic stability associated with conventional AOBs, this study optimized the preparation process and established a highly efficient and stable functional delivery system based on AOBs. This study employed single-factor experiments combined with the Box-Behnken response surface method to systematically optimize the preparation process of AOBs. Molecular docking was employed to predict the binding affinity between tea polyphenols (TP) and the structural protein Ctcaleosin of AOBs. Subsequently, the ultrasonic radical method was employed to modify and conjugate TP with AOB, thereby constructing functional AOBs mediated by tea polyphenol-protein complexes (AOB@TP); Network pharmacology was employed to systematically predict the targets for the synergistic obesity-alleviating effects of TP and medium-chain triglycerides (MCT) within AOB@TP. The preparation parameters were optimized as follows: 10 mg MCT, 100 μg egg yolk lecithin, and Ctcaleosin 275 μg. The binding rate of TP to AOBs prepared with the optimal parameters reached 94.55%, AOB@TP exhibited a spherical structure with a particle size of 216.67 nm, demonstrating excellent carrier properties; Network pharmacology predicted 67 potential targets for TP and MCT, with 60 overlapping targets related to obesity treatment, among which MMP9 and HIF1A were identified as core regulatory targets; GO functional enrichment analysis indicated that the targets were primary involved in biological processes such as lipid metabolism and energy homeostasis, while KEGG pathway enrichment primarily involved obesity-related signaling pathways. The AOB@TP functional delivery system AOB@TP developed in this study combines the advantages of small particle size and multifunctionality. It not only provides a novel, high-quality carrier for the efficient delivery of fat-soluble nutrients but also lays the theoretical and technical foundation for the in-depth application of functional AOBs in the intervention of metabolic diseases.