多酚氧化酶抑制剂高良姜素抗黑色素瘤机制研究

发布时间：2018-12-28 18:04

【摘要】：目的:研究高良姜素对多酚氧化酶(PPO)的抑制作用机制,并探讨了高良姜素与多酚氧化酶的结合机制,以期获得高良姜素抗人恶性黑色素瘤细胞株A375机理。方法:分光光度法检测高良姜素对PPO的抑制作用和抑制作用类型;荧光光谱法和分子对接法检测高良姜素与PPO酶的结合方式;MTT法测定高良姜素人恶性黑色素瘤细胞株A375增殖的抑制作用,Transwell法检测高良姜素对A375细胞侵袭的抑制作用。结果:高良姜素是一种竞争型的PPO抑制剂,对PPO氧化酶的半抑制率IC_(50)为(47.86±3.33)μmol/L,抑制常数Ki为(24.83±1.45)μmol/L;高良姜素能够有效猝灭PPO的荧光,高良姜素与PPO的结合常数Ka为(4.67±0.43)×10~4L/mol;高良姜素能够与PPO的活性中心铜离子发生结合作用,并与催化基团His259形成氢键;高良姜素能够明显的抑制A375细胞的增殖和转移,且细胞中的PPO活性和黑色素的合成量降低。结论:高良姜素是一个竞争性的PPO酶抑制剂,对调节黑色素水平起到重要作用,为临床抗皮肤癌提供了理论基础。
[Abstract]:Aim: to study the inhibitory mechanism of gingerin on polyphenol oxidase (PPO) and the mechanism of its binding with polyphenol oxidase (PPO) in order to obtain the mechanism of galangin against human malignant melanoma cell line A375. Methods: the inhibitory effects and types of gingerin on PPO were detected by spectrophotometry, and the binding modes of gingerin and PPO enzyme were detected by fluorescence spectrometry and molecular docking. The inhibitory effect of gingerin on the proliferation of human malignant melanoma cell line A375 was determined by MTT assay, and the inhibitory effect of galangin on the invasion of A375 cell line was detected by Transwell assay. Results: gingerin is a competitive PPO inhibitor. The semi-inhibitory rate of IC_ (50) on PPO oxidase is (47.86 卤3.33) 渭 mol/L, inhibition constant Ki is (24.83 卤1.45) 渭 mol/L;. Gingerin can effectively quench the fluorescence of PPO, and the binding constant Ka of gingerin to PPO is (4.67 卤0.43) 脳 10 ~ (4) L / mol, gingerin can bind to Cu ~ (2 +) ion in the active center of PPO and form hydrogen bond with the catalytic group His259. Gingerin significantly inhibited the proliferation and metastasis of A375 cells, and the PPO activity and melanin synthesis in A375 cells decreased. Conclusion: gingerin is a competitive inhibitor of PPO, which plays an important role in regulating melanin level and provides a theoretical basis for clinical anti-skin cancer.
【作者单位】：四川省医学科学院四川省人民医院皮肤外科;四川省医学科学院四川省人民医院肾脏内科;
【基金】：四川省卫计委科研课题(No.140084)
【分类号】：R739.5

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