基于代谢组学技术和QSAR模型的黑灵芝甾醇类化合物定量结构-活性关系研究

According to previous research, sterol compounds from Ganoderma have cholesterol lowering properties. However, the pathways of regulating cholesterol metabolism for sterol compounds from Ganoderma are still unclear. Besides, the relationship between the molecular structures of the sterol compounds and the hypocholesterolemic activity needs to be investigated..In the present project, high speed counter-current chromatography will be used in order to separate and purify enough sterol compounds from Ganoderma atrum for the investigation of their effects on cholesterol levels and metabolic profiles of hypercholesterolia Caco-2 and HepG2 cell model. A protocol for the metabolic profiling of Caco-2 and HepG2 cell lines will be developed based on HPLC-Q-TOF-MS techniques. Changes in metabolic profiles affected by sterol compounds from Ganoderma atrum will be observed using chemometrics methods, and the relationship between the molecular structures of the sterol compounds and the hypocholesterolemic activity will also be explored by QSAR models. The study may offer specific putative pathways in the cholesterol metabolism affected by sterol compounds.

现有研究表明，灵芝的主要活性成分甾醇类化合物具有明显的改善机体胆固醇水平作用，然而，灵芝中甾醇类化合物究竟与胆固醇代谢的哪些途径有关？各个途径调控胆固醇代谢的作用机制如何？甾醇类化合物的分子结构与调节胆固醇吸收、代谢相关活性之间的关系如何？这些问题尚不清楚，有待进一步研究。.本项目拟在前期工作基础上，采用高通量的高速逆流色谱制备分离技术对江西赣南地区特产的黑灵芝中的甾醇类化合物进行分离纯化，通过构建Caco-2 细胞胆固醇吸收和HepG2细胞胆固醇代谢模型，探明灵芝中甾醇类化合物对胆固醇吸收、代谢的影响，并采用代谢组学技术研究灵芝中甾醇类化合物调节胆固醇代谢的作用机制，在上述工作的基础上采用QSAR模型进一步探讨甾醇类化合物分子结构与调节胆固醇吸收、代谢相关活性之间的关系。该研究有助于深入理解甾醇类化合物降胆固醇的作用机制, 为甾醇类化合物生物活性的构效关系研究提供理论和实验依据。

本项目构建了HepG2细胞胆固醇代谢模型和Caco-2细胞吸收转运模型探究体外细胞中胆固醇的吸收和代谢，探明灵芝中甾醇类化合物（麦角甾醇、麦角甾醇酯、(22E)-麦角甾-4，6，8(14)，22-四烯-3酮和麦角甾-7，22-二烯-3β-醇）直接作用对胆固醇吸收和代谢的影响及作用机制及构效关系，并分析灵芝中甾醇类化合物对体外细胞内胆固醇吸收转运的调控机理，提出黑灵芝中甾醇类化合物直接调节胆固醇代谢的信号转导途径，并采用代谢组学技术研究灵芝中甾醇类化合物调节胆固醇代谢的作用机制。