基于生物活性和分子对接的滇鸡血藤水煎液质量控制研究

Study on Quality Control of Kadsurae Caulis Decoction Based on Biological Activity and Molecular Docking

目的:初步探究滇鸡血藤水煎液活血功效的生物评价方法并挖掘参与凝血酶抑制的核心成分。方法:采用体外凝血酶活性抑制试验,评价滇鸡血藤水煎液体外凝血酶抑制活性。然后利用超高效液相色谱-质谱(UPLC-MS)挖掘滇鸡血藤水煎液中的化学成分,并通过分子对接技术对这些化学成分的生物活性进行评估,筛选核心抗凝成分。结果:凝血酶抑制试验表明滇鸡血藤的标准煎剂(0.2 g·m L^(-1))具有高达78.40%的凝血酶抑制率,并从滇鸡血藤水煎液中推测鉴定了14个化合物,将其与凝血酶进行分子对接评价:戈米辛C(-7.9 kcal·mol^(-1))、戈米辛G(-7.6 kcal·mol^(-1))、内南五味子素C(-8.1kcal·mol^(-1))、内南五味子素D(-9.4 kcal·mol^(-1))、阿里山五味子灵C(-9.4 kcal·mol^(-1))、五味子素C(-7.9 kcal·mol^(-1))、五味子素A(-7.3 kcal·mol^(-1))、异型南五味子素E(-8.2 kcal·mol^(-1))、五味子酯D(-8.6 kcal·mol^(-1))、红花五味子素(-7.4 kcal·mol^(-1))、内南五味子酯B(-7.6 kcal·mol^(-1))、内南五味子酯A(-8.1 kcal·mol^(-1))、乙酰基氧代南五味子烷(-8.5 kcal·mol^(-1))、内南五味子酯D(-9.2 kcal·mol^(-1))。其中内南五味子素D和阿里山五味子灵C与凝血酶蛋白具有紧密结合的空间构型以及理想的结合能。结论:滇鸡血藤水煎液具有较好的凝血酶抑制活性,筛选出的2个具有凝血酶抑制活性的核心成分,可作为其质量标志物候选成分,为滇鸡血藤水煎液生物活性评价方法的建立和质控研究提供了参考依据。

Objective:To preliminary explore the biological evaluation method for the blood activating effect of kadsurae caulis decoction,and to explore the core components involved in thrombin inhibition.Methods:In vitro thrombin inhibition test was used to evaluate the thrombin inhibition activity of kadsurae caulis decoction.The chemical constituents in kadsurae caulis decoction were illuminated by UPLC-MS.The pharmacological activities of these compounds were evaluated by molecular docking,and potential thrombin activity inhibitors were screened from them.Results:The thrombin inhibition test showed that the thrombin inhibition rate of kadsurae caulis decoction is up to 78.40%,and 14 compounds were speculated and identified from the decoction by UPLC-MS,and their molecular docking with thrombin was evaluated:Gomisin C(-7.9 kcal·mol^(-1)),Gomisin G(-7.6 kcal·mol^(-1)),Interiorin C(-8.1 kcal·mol^(-1)),Interiorin D(-9.4 kcal·mol^(-1)),Schiarisanrin C(-9.4 kcal·mol^(-1)),Schisandrin C(-7.9 kcal·mol^(-1)),Schizandrin A(-7.3 kcal·mol^(-1)),Heteroclitin E(-8.2 kcal·mol^(-1)),Schisantherin D(-8.6 kcal·mol^(-1)),Rubschisandrin(-7.4 kcal·mol^(-1)),Interiotherin B(-7.6 kcal·mol^(-1)),Interiotherin A(-8.1 kcal·mol^(-1)),Acetoxyl Oxokadsurane(-8.5 kcal·mol^(-1)),Interiotherin D(-9.2 kcal·mol^(-1)).The docking results of these compounds indicated that Interiorin D and Schiarisanrin C had a highly compact spatial configuration and binding energy with thrombin protein.Conclusion:The thrombin inhibitory activity of kadsurae caulis decoction is excellent,and 2 core components with thrombin inhibitory activity have been screened out,that can be used as candidate components for the quality markers.This provides a reference basis for the establishment of a biological activity evaluation method and quality control research of kadsurae caulis decoction.