高山锥叶的成分分离鉴定及其胰脂肪酶抑制活性研究

Isolation and identification of constituents from the leaves of Castanopsis delavayi and their inhibitory activity against pancreatic lipase

关键词:  高山锥, 化学成分, 分离鉴定, 胰脂肪酶, 抑制活性

Key words:  Castanopsis delavayi, chemical constituents, isolation and identification, pancreatic lipase, inhibitory activity

摘要:

为系统探究高山锥叶中的化学成分,并明确其化合物对胰脂肪酶的抑制活性,该研究采用Sephadex LH-20、MCI gel CHP 20P、Diaion HP20SS、Toyopearl HW-40F等多种色谱柱交叉层析法,对高山锥叶80%甲醇提取物进行分离纯化,采用NMR、MS等波谱技术并结合文献对比鉴定化合物结构; 基于4-甲基伞形酮油酸酯(4-MUO)荧光测定法构建体外胰脂肪酶抑制活性筛选模型,以奥利司他为阳性对照(IC₅₀=0.001 4 mmol·L^-1),评价单体化合物的胰脂肪酶抑制活性。结果表明:(1)从高山锥叶80%甲醇提取物中共分离得到19个化合物,分别鉴定为没食子酸(1)、丁香酸(2)、顺式对羟基肉桂酸(3)、反式对羟基肉桂酸(4)、dehydrodigallic acid(5)、3-O-β-D-(6'-O-galloyl)-glucopyranoside(6)、龙胆酸5-O-β-D-( 6'-O-没食子酰基)-吡喃葡萄糖苷(7)、macarangioside E(8)、chesnatin(9)、茶倍素(10)、3-O-对香豆酰基奎宁酸(11)、5-O-咖啡酰基奎宁酸(12)、5-O-咖啡酰奎宁酸甲酯(13)、槲皮素(14)、杨梅素-3-O-葡萄糖苷(15)、槲皮素-3-O-β-D-半乳糖苷(16)、槲皮素-3-O-葡萄糖醛酸苷(17)、槲皮素-3-O-β-D-葡萄糖醛酸-6″-甲酯(18)、quercetin 3-O-(6″-O-galloyl)-β-D-galactopyranoside(hyperin 6″-gallate)(19)。(2)共筛选出12个对胰脂肪酶具有不同程度抑制活性的化合物,化合物6、7、12、17表现出较好的抑制活性。所有化合物均为首次从高山锥中分离得到,该研究为高山锥降脂作用的深入研究提供了理论依据。

Abstract:

A phytochemical investigation of the leaves of Castanopsis delavayi was conducted to isolate and characterize its constituent compounds and evaluate their inhibitory activity against pancreatic lipase. The 80% methanol extract of the leaves was sequentially purified using multiple chromatographic columns, including Sephadex LH-20, MCI gel CHP 20P, Diaion HP20SS, and Toyopearl HW-40F. The structure of isolated compounds was identified by pop analysis(NMR, MS)in comparison with literature data. An in vitro screening model for pancreatic lipase inhibitory activity was established utilizing the 4-methylumbelliferyl oleate(4-MUO)fluorescence assay, with orlistat serving as the positive control(IC₅₀=0.001 4 mmol·L^-1). The results were as follows:(1)Nineteen compounds were successfully isolated and structurally characterized from the 80% methanol extract: gallic acid(1), syringic acid(2),(Z)-3, 5-dihydroxycinnamic acid(3), (E)-3, 5-dihydroxycinnamic acid(4), dehydrodigallic acid(5), 3-O-β-D-(6'-O-galloyl)-glucopyranoside(6), gentisic acid-5-O-β-D-(6'-O-galloyl)-glucopyranoside(7), macarangioside E(8), chesnatin(9), theogallin(10), 3-O-p-coumaroylquinic acid(11), 5-O-caffeoylquinic acid(12), methyl chlorogenate(13), quercetin(14), myricetin-3-O-β-D-glucopyranoside(15), quercetin-3-O-β-D-galactoside(16), quercetin-3-O-glucuronopyranoside(17), quercetin-3-O-β-D-glucuronide-6″-methyl ester(18), and quercetin 3-O-(6″-O-galloyl)-β-D-galactopyranoside(hyperin 6″-gallate)(19).(2)Twelve compounds exhibiting differential inhibitory effects against pancreatic lipase, which compounds 6, 7, 12, and 17 demonstrated significant inhibitory activity. All compounds were isolated from C. delavayi for the first time, providing a theoretical basis for further research on its lipid-lowering effects.