凤冈富锌硒茶对ERp57 蛋白酶的抑制作用及其有效成分的虚拟筛选

Inhibitory effect of Se-enriched and zinc-enriched green tea of Fenggang on ERp57 protease and virtual screening of its effective constituents

摘要:

目的 探讨凤冈富锌硒茶水提取物对抗血小板聚集的潜在药物靶点ERp57 蛋白酶的抑制作用及 其潜在的有效成分。方法 利用超高效液相色谱分析凤冈富锌硒茶水提取物的主要成分，采用胰岛素浑浊度 分析方法检测提取物对ERp57 蛋白酶的抑制作用，采用体外血小板聚集率实验分析提取物对血小板聚集的干 预作用。采用计算机虚拟筛选和实验验证相结合的方法，分析提取物中抑制ERp57 蛋白酶的主要活性成分。 结果 凤冈富锌硒茶水提取物主要存在4 种化合物，提取物对ERp57 蛋白酶有显著的抑制作用，且表现出一 定程度的抗血小板聚集作用，其中表没食子儿茶素没食子酸酯（EGCG）对ERp57 蛋白酶的抑制作用最强。 结论 长期饮用绿茶有益于降低心血管疾病死亡率，可能与绿茶抑制ERp57 蛋白酶活性，从而抗血小板聚集 有关，其中EGCG 是其主要有效成分之一。

Abstract:

Objective To examine the effect of Se-enriched and zinc-enriched green tea of Fenggang on the enzymatic activity of ERp57, a potential anti-platelet target, and try to clarify the major constituents of the extract. Methods Ultra performance liquid chromatography (UPLC) was used to assay the major components of the green tea. The effect of the extract on platelet function and ERp57 activity was evaluated by light transmission aggregometry and insulin reduction assay, respectively. The major active constituents of the green tea were identified using virtual screening and experimental validation. Results Using UPLC, 4 major components were identified from the tea extract. Further experiments with insulin turbidity assay and virtual screening using computer-based methods demonstrated the green tea extract displayed potent inhibitory effect on the enzymatic activity of ERp57 (among which EGCG had the strongest effect) and certain anti-platelet aggregation effect. Conclusions The beneficial effect of green tea on the mortality of CVD patients could be associated with its inhibition of ERp57 activity and platelet aggregation. In addition, EGCG is one of the major active constituents of the green tea.