摘要: |
珠子参叶是五加科植物珠子参Panax japonicas C. A. Mey. var. major (Burk.) C.Y.Wu et K.M.Feng的干燥带梗叶。为了合理开发利用珠子参叶,本研究利用HPLC方法分析珠子参叶皂苷部位的化学成分,测定珠子参叶皂苷部位的脂肪酶抑制活性,并利用酶动力学测定参叶皂苷部位对脂肪酶的抑制类型,通过分子对接辅助探讨脂肪酶抑制机制,最后利用高血脂症小鼠模型验证珠子参叶皂苷部位对小鼠血清中胆固醇和甘油三脂含量的影响。结果表明:(1)从珠子参叶皂苷部位中鉴定6个皂苷类化合物,分别为20(S)-人参皂苷Rg2、20(R)-人参皂苷Rg2、人参皂苷Rb2、人参皂苷Rb3、人参皂苷Rd、人参皂苷Rh2;(2)脂肪酶抑制活性的测试结果表明珠子参叶皂苷部位、20(R)-人参皂苷 Rg2对脂肪酶具有较强的抑制作用,其IC50分别为0.14和2.30 μmol/L;(3)动力学结果表明珠子参叶皂苷部位、20(R)-人参皂苷Rg2、人参皂苷Rb3对脂肪酶的抑制为可逆性抑制,抑制类型为非竞争型抑制;(4)分子对接研究显示:配体与ARG337B、ASP331B、ILE248B残基结合可能有助于提高配体的脂肪酶抑制活性;(5)动物实验证明珠子参叶总皂苷可以显著降低高脂血症小鼠血清中胆固醇和甘油三脂的含量。该研究为珠子参叶在降血脂方面的深入开发利用奠定基础。 |
关键词: 珠子参叶,皂苷,脂肪酶,酶动力学,分子对接,体内验证 |
DOI: |
分类号:R932 |
基金项目:陕西省科技厅项目(2018ZDXM-SF-007);陕西中医药大学创新团队项目(2019-YL12);陕西中医药大学质量提升工程项目(ZG031)[Supported byShaanxi Provincial Science and Technology Department Project (2018ZDXM-SF-007);Innovation Team Project of Shaanxi University of Traditional Chinese Medicine (2019-YL12);Shaanxi University of Traditional Chinese Medicine Quality Improvement Project ( ZG031 )] |
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Study on the mechanism of lipase inhibition and hypolipidemic effect of Panax japonicas leaves |
ZHENG Huachan, MENG Lingru, HE Miao, HUANG Wenli, DAN Linwei, XU Hong, DENG Chong, ZHANG Huawei, JIANG Yi, SONG Xiaomei
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School of Pharmacy,Shaanxi University of Chinese Medicine
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Abstract: |
The leaves of Panax japonicas C. A. Mey. var. major (Burk.) C.Y. Wu et K. M. Feng ( Araliaceae ) are dry sessile leaves. In order to rationally develop and utilize the leaves of Panax japonicas, the HPLC method was used to analysis and identify the saponins from the leaves of Panax japonicas. The inhibitory activity and the inhibitory type of the saponins from the leaves of Panax japonicas on lipase was determined with enzyme kinetics. The inhibition mechanism of lipase was investigated with molecular docking. Finally, the effect of saponins from the leaves of Panax japonicas on the content of cholesterol and triglyceride in serum of mice was verified in hyperlipidemia mouse model. The results showed that: (1) Six compounds were identified from the saponins of Panax japonicas leaves, which were 20 (S) -Ginsenoside Rg2, 20 (R) -Ginsenoside Rg2, Ginsenoside Rb2, Ginsenoside Rb3, Ginsenoside Rd and Ginsenoside Rh2. (2) The results of lipase inhibitory activity showed that 20 (R) -Ginsenoside Rg2 had strong inhibitory effect on lipase with IC50 of 0.14 and 2.30 μmol/L respectively; (3) The kinetic results exerted that the lipase activity of Panax japonicas leaf saponins, 20 (R) -Ginsenoside Rg2 and Ginsenoside Rb3 were all reversible inhibition, and the inhibition type was non-competitive inhibition. (4) Molecular docking studies showed that the binding of ligands to ARG337B, ASP331B and ILE248B residues may help to improve the lipase inhibitory activity of ligands. (5) the results of animal experiments suggested that total saponins of Panax japonicas leaves could significantly reduce the content of cholesterol and triglyceride in serum of hyperlipidemia mice. This study laid a foundation for the further development and utilization of Panax japonicas leaves. |
Key words: Panax japonicas leaf, Saponin, Pancreatic lipase, Enzyme kinetics, Molecular docking, Verification in vivo |