黄柏碱对α-葡萄糖苷酶的体外抑制作用

郑丽婷; 周鸿; 刘奕明; 林爱华

黄柏碱对α-葡萄糖苷酶的体外抑制作用

1.
广州中医药大学第二附属医院，广东广州 510120
2.
广东省中医证候临床研究重点实验室，广东广州 510120

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出版历程
- 刊出日期: 2020-11-10

Inhibitory Effect of Phellodendrine on α-Glucosidasein vitro

1.
The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510120, China
2.
Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome, Guangzhou, 510120, China

摘要

摘要: 目的研究黄柏碱对α-葡萄糖苷酶的体外抑制作用、作用类型和分子机制。方法建立α-葡萄糖苷酶抑制剂体外筛选模型，测定黄柏碱对α-葡萄糖苷酶的抑制率，并采用动力学方法研究黄柏碱的酶抑制作用类型。采用同源模建的方法构建酿酒酵母α-葡萄糖苷酶的三维结构，并运用分子对接技术分析黄柏碱抑制α-葡萄糖苷酶的分子作用机制。结果黄柏碱对α-葡萄糖苷酶的半数抑制浓度（IC₅₀）为126.36 mg/L，且抑制率随浓度增加而增加。酶动力学结果显示，黄柏碱浓度增大，反应速率降低，V_max、K_m变小。分子对接结果显示，黄柏碱与α-葡萄糖苷酶位点5的结合能最低且其最好的对接构象结合能为-31.4 kJ/mol。黄柏碱与α-葡萄糖苷酶分子中的LYS15、SER295、HIS258等氨基酸残基形成氢键，与残基ALA289形成疏水相互作用以及与残基GLU10形成π-阴离子相互作用。结论黄柏碱是α-葡萄糖苷酶的可逆性反竞争性抑制剂，氢键、疏水作用和π-阴离子相互作用是黄柏碱与α-葡萄糖苷酶分子间的主要作用力。
- 黄柏碱 /
- α-葡萄糖苷酶 /
- 反竞争性抑制 /
- 同源建模 /
- 分子对接
Abstract: OBJECTIVE To study the inhibitory effect and the inhibition type of phellodendrine on α-glucosidasein vitro and explore the molecular mechanism. METHODS An inhibitor screening model was establishedin vitro to examine the inhibitory rate of phellodendrine on α-glucosidase.The inhibition type was investigated by kinetic method. The three-dimensional structure of α-glucosidase （Saccharomyces cerevisiae） was constructed by homology modeling method， and the inhibition molecular mechanism of phellodendrine on α-glucosidase was analyzed by molecular docking technology. RESULTS Phellodendrine displayed obvious inhibitory activity on α-glucosidase with an IC₅₀ value of 126.36 mg/L， in a concentration-dependent manner. The results of enzyme kinetics indicated that the reaction rate was lowered and V_max， K_m were decreased with the concentration increasing of phellodendrine. The results of molecular docking showed that the binding energy of phellodendrine to α-glucosidase site 5 was the lowest， and its optimal docking conformation binding energy was -31.4 kJ/mol. Phellodendrine exerted its inhibitory activity by forming hydrogen bonds with amino acid residues such as LYS15， SER295， HIS258， hydrophobic interaction with ALA289 and π-anion interaction with GLU10. CONCLUSION Phellodendrine is a reversible uncompetitive inhibitor of α-glucosidase. The hydrogen bond， hydrophobic interaction and π-anion interaction are main forces between phellodendrine and α-glucosidase.
- phellodendrine /
- α-glucosidase /
- uncompetitive inhibition /
- homology modeling /
- molecular docking

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