基于UFLC-MS/MS结合网络药理学及实验验证的乌药叶抗高脂血症作用及机制研究  

Research on the Anti-hyperlipidemia Effect and Mechanism of Linderae Folium via UFLC-MS/MS Combined with Network Pharmacology and Experimental Validation

作　　者：蔡中齐 罗益远 王芙蓉 李娜 蔡红蝶 陈妮妮 陈宏降 王娟 CAI Zhongqi;LUO Yiyuan;WANG Furong;LI Na;CAI Hongdie;CHEN Nini;CHEN Hongjiang;WANG Juan(Zhejiang Pharmaceutical University,Ningbo 315100,China)

机构地区：[1]浙江药科职业大学,浙江宁波315100

出　　处：《中国现代应用药学》2024年第24期3553-3564,共12页Chinese Journal of Modern Applied Pharmacy

基　　金：浙江省医药卫生科技计划项目(2023KY298);浙江省基础公益研究计划项目(LTGN23H280002);浙江省大学生科技创新活动计划暨新苗人才计划项目(2022R460A001);宁波市自然科学基金项目(2022J194,2023J306,2023J309);宁波市科技计划项目(2023S187);浙江药科职业大学校级科研课题(2022134,2022135)。

摘　　要：目的采用UFLC-MS/MS技术分析乌药叶中化学成分,依据解析的化学成分结合网络药理学、分子对接技术和动物实验验证分析乌药叶防治高脂血症的主要活性成分、作用靶点及相关通路,初步探索其防治高脂血症的潜在作用机制。方法通过UFLC-MS/MS技术分析乌药叶中化学成分,采用网络药理学对高脂血症相关靶点进行预测,构建“化合物-疾病-靶点”网络。通过GO功能和KEGG分析,筛选出显著富集的通路(P<0.05)。对筛选出的活性成分与高脂血症关键靶点进行分子对接验证,最后采用动物实验和RT-PCR技术对乌药叶药效和抗高脂血症关键靶点进行实验验证。结果共分析鉴定32个化合物,其中包括8个生物碱类、16个黄酮类、8个内酯类化合物。网络药理学研究共筛选出活性成分13个,涉及相关基因靶点46个。基于GO和KEGG分析乌药叶降脂作用的潜在机制,主要涉及以下几条通路:流体剪应力与动脉粥样硬化信号通路、AGE-RAGE信号通路、TNF信号通路、脂肪细胞因子信号通路等。动物实验表明,乌药叶醇提物2个剂量均可明显调节模型小鼠血脂、血糖和肝脏脂质水平;且RT-PCR结果显示其可明显上调肝脏组织中清道夫受体B类Ⅰ型(scavenger receptor class B typeⅠ,SR-BI)的表达,下调酰基辅酶A-胆固醇酰基转移酶(acyl-coenzyme A:cholesterol acyltransferase,ACAT)基因的表达。结论乌药叶具有改善高脂血症作用,其通过多成分、多靶点、多通路发挥效应,潜在活性成分排名前5的为槲皮素、山柰酚、四氢小檗碱、波尔定碱和四氢非洲防己碱,它们可能通过调节动脉粥样硬化、脂肪细胞因子等信号通路参与炎症反应、脂肪代谢、胰岛素抵抗和血管内皮功能等过程发挥降脂作用机制。OBJECTIVE To explore the potential mechanism of Linderae Folium on prevention and treatment of hyperlipidemia,UFLC-MS/MS was used to analyze the chemical components of Linderae Folium,then the main active components,targets and related pathways of Linderae Folium against hyperlipidemia were verified by the chemical composition,network pharmacology,molecular docking technology and animal experiments.METHODS The chemical constituents in the Linderae Folium were analyzed by UFLC-MS/MS,and the targets related to hyperlipidemia were predicted by network pharmacology,and a"compound-disease-target"network was constructed.Through GO function and KEGG analysis,the significantly enriched pathways were screened(P<0.05).The selected active ingredients and the key targets of hyperlipidemia were verified by molecular docking.Finally,animal experiments and RT-PCR were used to verify the efficacy and anti-hyperlipidemia key targets.RESULTS There were 32 chemical components analyzed by qualitative analysis of Linderae Folium,including 8 alkaloids,16 flavonoids and 8 lactones.A total of 13 active ingredients and 46 related gene targets were screened by network pharmacological study.To explore the potential mechanism of Lindera Folium on lipid-lowering effect based on GO and KEGG analysis,the results suggested that the mechanism was mainly involved in the following pathways:fluid shear stress and atherosclerosis,AGE-RAGE signaling pathway,TNF signaling pathway,adipocytokine signaling pathway and so on.Animal experiments showed that the 2 doses of alcohol extract of Linderae Folium could significantly regulate the plasma lipid,blood glucose,and liver lipid levels of hyperlipidemia mice.The results of RT-PCR showed that Lindera Folium could significantly upregulate the expression of scavenger receptor class B typeⅠ(SR-BI)mRNA,and downregulate the level of acyl-coenzyme A:cholesterol acyltransferase(ACAT)mRNA in liver.CONCLUSION Lindera Folium might have effective on alleviate hyperlipidemia,and it exerted multi-component,multi-targ

关 键 词：乌药叶 UFLC-MS/MS 高脂血症 网络药理学 作用机制 

分 类 号：R285.5[医药卫生—中药学]