山楂依赖肠道菌群通过卵磷脂代谢调脂的作用研究

Dyslipidemia and atherosclerosis sex heart disease and its complications are closely related, hawthorn has the lipid effect.The latest research has found that，gut flora plays an crucial role in the generation and progression of atherosclerosis induced by abnormal phosphatidylcholine metabolism. in previous works, we hypothesis that one of the significant pathways to regulate lipid of Crataegus pinnatifida Bge is through gut-flora-dependent metabolism of phosphatidylcholine”. Using the Apoe-/- mice on diets supplemented with choline induced abnormal phosphatidylcholine metabolism model to investigate the regulation pathway of Crataegus pinnatifida Bge in atherosclerosis promoted by abnormal phosphatidylcholine metabolism via gut flora. ① Isotopic tracer method is used to analyze the correlation between Crataegus pinnatifida Bge and gut flora. ② Flow cytometry analysis is performed to prove that Crataegus pinnatifida Bge be able to inhibit peritoneal macrophages cholesterol accumulation and foam cell formation which included suppressing the gut flora, significantly inhibiting dietary-choline-induced macrophage foam cell formation and declining macrophage scavenger receptor expression. ③ RT-PCR method is performed to discover Crataegus pinnatifida Bge preparations’ inhibition of both mRNA levels and surface protein levels of macrophage scavenger receptors are correlated with suppression of gut flora. The aim are to clarify the molecular mechanism of abnormal phosphatidylcholine metabolism induced atherosclerosis during treatment of Crataegus pinnatifida Bge and paves the way for formation of new theory in prevention and treatment of atherosclerotic heart disease.

血脂异常与动脉粥样硬化（AS）性心脏病及其并发症密切相关，山楂具有调脂功效。最新研究发现，肠道微生物在卵磷脂代谢异常致AS发生发展过程中起着重要作用。本项目在前期工作基础上，提出“山楂通过肠道菌群参与的卵磷脂代谢是其调脂的一个重要途径”假说，研究山楂对肠道菌群参与的卵磷脂代谢异常所致AS的调节途径，采用载脂蛋白E基因敲除小鼠，胆碱强化饮食造成卵磷脂代谢异常小鼠动物模型。通过①同位素示踪法等观察山楂与肠道菌群间联系。②流式细胞术检测等证明山楂能够抑制腹腔巨噬细胞形成泡沫细胞，包括抑制肠道菌群，明显的减少膳食卵磷脂诱导的泡沫细胞形成和下调巨噬细胞受体表达的水平。③RT-PCR等探讨山楂制剂抑制巨噬细胞清道夫受体mRNA和表面蛋白水平与抑制肠道菌群相关联。旨在阐明山楂治疗卵磷脂代谢异常所致动脉粥样硬化的分子机制，为山楂防治AS性心脏病及其并发症提供新的理论基础，为中药调脂阐明一条新的途径。

动脉粥样硬化与血脂异常及卵磷脂代谢异常密切相关，而肠道菌群的功能则影响着饮食卵磷脂转化生成TMAO、巨噬细胞胆固醇的积累和泡沫细胞的形成。本课题以山楂和肠道菌群调控的腹腔巨噬细胞受体表达作为切入点来开展体内外研究。. 本课题研究山楂对肠道菌群参与的卵磷脂代谢异常所致AS的调节途径，采用载脂蛋白E基因敲除小鼠，胆碱强化饮食造成卵磷脂代谢异常小鼠动物模型，①观察山楂通过肠道菌群治疗脂质代谢异常；②流式细胞术检测等证明山楂能够抑制腹腔巨噬细胞形成泡沫细胞，包括抑制肠道菌群，明显的减少膳食卵磷脂诱导的泡沫细胞形成和下调巨噬细胞受体表达的水平；RT-PCR等探讨山楂制剂抑制巨噬细胞清道夫受体mRNA和表面蛋白水平与抑制肠道菌群相关联。旨在阐明山楂治疗卵磷脂代谢异常所致动脉粥样硬化的分子机制，为山楂防治AS性心脏病及其并发症提供新的理论基础，为中药调脂阐明一条新的途径。. 实验结果表明，山楂能够减少高胆碱饮食喂养造成动脉粥样硬化模型小鼠病变处形成的细胞炎症反应，能够显著降低巨噬细胞CD36 mRNA和SR-A1 mRNA的表达水平，抑制腹腔巨噬细胞中CD36，SR-A1的蛋白表达水平，增强机体的抗氧化能力,有效改善血脂，山楂制剂改善血脂可能与肠道菌群相关。山楂叶提取物金丝桃苷（Hyp）具有抗LPS诱导的RAW264.7细胞炎症反应的作用，其作用可能与调控p38MAPK/Sirt6/NF-κB信号通路相关。Hyp能够抑制腹腔巨噬细胞形成泡沫细胞，包括抑制肠道菌群，明显的减少膳食卵磷脂诱导的泡沫细胞形成和下调巨噬细胞受体表达的水平；抑制巨噬细胞清道夫受体mRNA和表面蛋白水平与抑制肠道菌群相关联。本研究为山楂防治AS性心脏病及其并发症提供新的理论基础，为中药调脂阐明一条新的途径。