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目的从分子水平探讨乙酰泽泻醇降低胆固醇(TC)的作用机理。方法利用试剂盒法测定了调脂中药泽泻主要有效成分23-乙酰泽泻醇B和24-乙酰泽泻醇A对高脂小鼠TC的影响,利用试剂盒法、Western blotting技术结合分子模拟技术研究两者对TC代谢关键酶3-羟基-3-甲基戊二酸单酰辅酶A(HMG-Co A)还原酶作用的分子机理。结果乙酰泽泻醇能显著降低高脂小鼠TC(P<0.01,P<0.05),23-乙酰泽泻醇B作用强度高于24-乙酰泽泻醇A(P<0.05),同时在体内、体外均可剂量依赖性下调HMG-Co A还原酶活性(P<0.01),且23-乙酰泽泻醇B对该酶作用强于24-乙酰泽泻醇A(P<0.05)。两者均未显著下调HMG-Co A还原酶的蛋白表达(P>0.05)。两者与HMG-Co A还原酶结合的关键氨基酸残基可能为Lys691、Asp767、Asn658。结论乙酰泽泻醇降低TC的机制可能是其原型药物通过抑制HMG-Co A还原酶活性来达到,且可能是通过直接竞争性与HMG-Co A还原酶结合抑制其作用,乙酰泽泻醇的舵手基团为其侧链,侧链与母环折叠结合弱,打开结合强。
Objective To investigate the mechanism of acetyl-alisol-lowering cholesterol (TC) at the molecular level. Methods The effects of 23-acetyl alisol B and 24-acetyl alisol A, the major active constituents of Alismataceae, a lipid-lowering herbal medicine, on the TC of hyperlipidemic mice were determined by the kit method. Using the kit method, Western blotting and molecular modeling The molecular mechanism of their interaction on HMG-Co A reductase, a key enzyme in TC metabolism, was studied. Results Acetisine Alisol could significantly decrease the TC of high-fat mice (P <0.01, P <0.05). The intensity of 23-acetyl alisol B was higher than that of 24-acetyl alisol A (P <0.05) In vitro, HMG-Co A reductase activity was down-regulated in a dose-dependent manner (P <0.01), and 23-acetyl alisol B had a stronger effect on this enzyme than 24-acetyl alisol A (P <0.05). Both did not significantly down-regulate the protein expression of HMG-Co A reductase (P> 0.05). The two key amino acid residues that bind to HMG-Co A reductase may be Lys691, Asp767, Asn658. Conclusions Acetose alisol may decrease the TC level by inhibiting the activity of HMG-Co A reductase, possibly through the direct competitive inhibition of HMG-Co A reductase, The helmsman group is its side chain, the side chain and the mother ring are folded and combined weakly, and the bond is strong.