大豆苷在大鼠肝、肾、小肠黏膜匀浆中的酶解研究  

Enzymolysis of daidzin in rat liver,kidney and small intestine mucosal homogenates

作　　者：夏婷 王新雨 张锋林 谭晓梅 张艳华 XIA Ting;WANG Xin-yu;ZHANG Feng-lin;TAN Xiao-mei;ZHANG Yan-hua(Guangdong Provincial Key Laboratory for Chinese Medicine Pharmaceutics,Guangdong Provincial Engineering Laboratory for Preparation Technology of Chinese Medicine/College of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,China;Guangxi Huahong Pharmaceutical Co.,Ltd.,Liuzhou 545199,China)

机构地区：[1]南方医科大学中医药学院,广东省中药制剂重点实验室,广东省中药制剂技术工程实验室,广东广州510515 [2]广西壮族自治区花红药业集团股份公司,广西柳州545199

出　　处：《中成药》2023年第3期708-713,共6页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金(81973489);广东省自然科学基金(2019A1515011398)。

摘　　要：目的考察大豆苷在大鼠肝、肾、小肠黏膜匀浆中的酶解。方法UPLC-MS/MS法测定大豆苷在肝、肾、小肠黏膜匀浆中的含量,计算K_(m)、V_(max)。分别采用D-葡萄糖酸-1,5-内酯、D-葡萄糖二酸-1,4-内酯验证乳糖酶根皮苷水解酶、β-葡萄糖醛酸苷酶是否在肾中水解大豆苷。再对大豆苷、β-葡萄糖醛酸苷酶进行分子对接。结果大豆苷在肝匀浆中不水解,在肾、小肠黏膜匀浆中水解,并且在后者中程度更高。大豆苷在肾匀浆中的代谢符合典型酶动力学过程,而在小肠黏膜匀浆中的代谢符合非典型酶动力学过程。β-葡萄糖醛酸苷酶活性被抑制后,大豆苷代谢率降低;乳糖酶根皮苷水解酶活性被抑制后,大豆苷元生成速率降低。β-葡萄糖醛酸苷酶可与大豆苷直接结合。结论大豆苷在大鼠肾匀浆中可高效转化为大豆苷元,肾、小肠黏膜中其酶解动力学存在差异。β-葡萄糖醛酸苷酶、乳糖酶根皮苷水解酶均参与大豆苷转化。AIM To investigate the enzymolysis of daidzin in rat liver,kidney and small intestine mucosal homogenates.METHODS UPLC-MS/MS was adopted in the content determination of daidzin in liver,kidney and small intestine mucosal homogenates,after which K_(m)and V_(max)were calculated.D-glucuronide 1,5-lactone and D-glucuronide 1,4-lactone were applied to verifying whether lactase phlorizin hydrolase and β-glucuronidase hydrolysed daidzin in kidney,respectively.Subsequently,molecular docking was performed inβ-glucuronidase and daidzin.RESULTS Daidzin did not hydrolyze in liver homogenate,but hydrolyzed in kidney homogenate and small intestinal mucosa homogenate,whose degree was higher in the latter.The metabolism of daidzin in kidney homogenate accorded with typical enzyme kinetic process,while that in small intestinal mucosal homogenate accorded with atypical enzyme kinetic process.After β-glucuronidase and lactase phlorizin hydrolase activities were inhibited,the metabolic rate and production rate of daidzin were decreased,respectively.β-Glucuronidase could directly bind to daidzin.CONCLUSION Daidzin can be efficiently converted into daidzein in rat kidney homogenate,whose enzymolysis kinetics in kidney and small intestine mucosa exist differences.Both β-glucuronidase and lactase phlorizin hydrolase are involved in the conversion process of daidzin.

关 键 词：大豆苷 酶解 肝匀浆 肾匀浆 小肠黏膜匀浆 UPLC-MS/MS 分子对接 

分 类 号：Q814[生物学—生物工程]