基于药效团模型的龙血竭潜在降糖活性成分筛选  

Screening of potential hypoglycemic active ingredients from Resina Draconi based on pharmacophore model

作　　者：莫钧茹 覃淼 朱素梅 黄金梅 奉建芳 黎芳[1,3] 梁健钦 MO Jun-ru;QIN Miao;ZHU Su-mei;HUANG Jin-mei;FENG Jian-fang;LI Fang;LIANG Jian-qin(Guangxi University of Chinese Medicine,Nanning 530200,China;Guangxi Superior Proprietary Chinese Medicine and Ethnic Medicine Development Engineering Technology Research Center,Nanning 530023,China;Guangxi Key Laboratory of Common Technology of Traditional Chinese Medicine Preparation in Laboratory,Nanning 530200,China)

机构地区：[1]广西中医药大学,广西南宁530200 [2]广西优势中成药与民族药开发工程技术研究中心,广西南宁530023 [3]广西中药制剂共性技术研发重点实验室,广西南宁530200

出　　处：《中草药》2022年第15期4764-4772,共9页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(81960872);广西研究生教育计划项目(YCSW2021240)。

摘　　要：目的 基于药效团模型筛选龙血竭Resina Draconi潜在降糖活性成分。方法 采用葡萄糖氧化酶法测定龙血竭含药血清对HepG2细胞胰岛素抵抗模型的葡萄糖消耗量。通过液相色谱-质谱联用技术(liquid chromatography-mass spectrometry,LC-MS)鉴定和检索中国知网(CNKI)、Web of Science数据库建立龙血竭的化合物库,通过Discovery Studio的吸收、分布、代谢、排泄、毒性(absorption,distribution,metabolism,excretion,toxicity,ADMET)模块预过滤化合物,通过钠-葡萄糖协同转运蛋白2(sodium-glucose cotransporter 2,SGLT2)、二肽基肽酶4(dipeptidyl peptidase 4,DPP4)药效团模型和分子对接筛选候选有效成分,通过细胞实验验证候选有效成分的降糖活性。结果 与模型组比较,龙血竭含药血清能使胰岛素抵抗的HepG2细胞葡萄糖消耗量显著增加(P<0.05)。经ADMET模块筛选到179个化合物,经最优药效团模型SGLT2-02、DPP4-03从中筛选出SGLT2、DPP4潜在抑制剂14个,其中5’-甲氧基松脂素(medioresinol)、开环异落叶松树脂酚[(+)-secoisolariciresinol]、1-palmitoylglycerophosphocholine、L-茶氨酸(L-theanine)的结合能均优于对照配体,分别为-852.285、-798.060、-916.116、-667.356 kJ/mol。与模型组相比,开环异落叶松树脂酚、L-茶氨酸能显著提高HepG2细胞葡萄糖消耗量(P<0.05、0.01)。结论 体外实验证实龙血竭显示良好的降糖活性,龙血竭中的5’-甲氧基松脂素、开环异落叶松树脂酚是SGLT2的潜在抑制剂,1-palmitoylglycerophosphocholine、L-茶氨酸是DPP4的潜在抑制剂,开环异落叶松树脂酚、L-茶氨酸具有降糖活性。Objective To screen potential hypoglycemic active ingredients of Longxuejie(Resina Draconis, Chinese dragon’s blood)based on pharmacophore model. Methods The glucose consumption of serum containing Resina Draconis on insulin resistance model of HepG2 cells was determined by glucose oxidase method. The compound database of Resina Draconis was established by liquid chromatography-mass spectrometry(LC-MS) identify and searching CNKI and Web of Science database. Then compounds were filtered by ADMET(absorption, distribution, metabolism, excretion and toxicity) module of Discovery Studio. The candidate active ingredients were screened by pharmacophore models for sodium-glucose cotransporter 2(SGLT2) and dipeptidyl peptidase 4(DPP4)and molecular docking. Finally, the activity of candidate active ingredients were verified by cellular experiments. Results The serum containing Resina Draconis significantly increased glucose consumption of insulin-resistant HepG2 cells compared with the model group(P < 0.05). A total of 179 compounds from Resina Draconis were screened by ADMET module. A total of 14 potential inhibitors of SGLT2 and DPP4 were screened by the optimal pharmacophore model of SGLT2-02 and DPP4-03. The binding energies of medioresinol,(+)-secoisolariciresinol, 1-palmitoylglycerophosphocholine and L-theanine were lower than the control ligands, which were-852.285,-798.060,-916.116,-667.356 kJ/mol, respectively.(+)-Secoisolariciresinol and L-theanine could significant increase glucose consumption of HepG2 cells compared with the model group(P < 0.05, 0.01). Conclusion Resina Draconis showed good in vitro hypoglycemic activity. Medioresinol and(+)-secoisolariciresinol were the potential inhibitors of SGLT2, and 1-palmitoylglycerophosphocholine and L-theanine were the potential inhibitors of DPP4.(+)-Secoisolariciresino and L-theanine showed good in vitro hypoglycemic activity.

关 键 词：龙血竭 降糖 ADMET 药效团 分子对接 钠-葡萄糖协同转运蛋白2 二肽基肽酶4 开环异落叶松树脂酚 L-茶氨酸 

分 类 号：R284[医药卫生—中药学] R285[医药卫生—中医学]