基于HPLC-QTOF/MS及G蛋白偶联受体分析的延胡索物质基础及作用机制研究

Chemical constituents and mechanism of Corydalis Rhizoma based on HPLC-QTOF/MS and G protein-coupled receptor analysis

为明确延胡索药材的物质基础并初步探讨其作用机制,本研究利用高效液相色谱-四级杆/飞行时间质谱(HPLC-QTOF/MS)对延胡索药材的60%乙醇提取物的化学成分进行分析鉴定,并通过胞内钙离子浓度荧光检测法考察延胡索药材提取物及其代表性单体成分原阿片碱、巴马汀和延胡索乙素对6个G蛋白偶联受体即5-羟色胺受体(5-HT_(1A))、μ阿片受体(OPRM1)、β_2肾上腺素受体(ADRB_2)、多巴胺受体(D_2)、乙酰胆碱受体(M_2)和血栓素-前列腺素受体(TP)的激动和拮抗作用。实验共分析得到延胡索药材提取物中的31个化合物,鉴定出28个生物碱类成分。G蛋白偶联受体实验表明,延胡索提取物能激动5-HT1A、OPRM1和ADRB2受体,拮抗D2受体而发挥生物活性;原阿片碱只对D_2、M_2受体显示拮抗作用;延胡索乙素可激动ADRB2受体,拮抗D_2、TP受体;而巴马汀对6个受体无显著作用。结果体现了延胡索多成分、多靶点、多途径发挥药效的作用机制。

The chemical constituents of Corydalis Rhizoma were identified in the 60% ethanol extract using high performance liquid chromatography/electrospray ionization quadruple time-of-flight mass spectrometry（HPLC-QTOF/MS）. The stimulation and inhibition effects of Corydalis Rhizoma and its representative compounds（protopine, palmatine, tetrahydropalmatine） on G protein-coupled receptor（GPCR）, including 5-hydroxytryptamine 1A receptor（5-HT_（1A））, μ opioid receptor（OPRM1）, β_2 adrenergic receptor（ADRB2）, dopamine receptor（D_2）, acetylcholine receptor（M_2） and thromboxane-prostaglandin receptor（TP） were explored using the fluorescence assay of intracellular calcium ion. As a result, 31 compounds were obtained and 28 alkaloid compounds were identified. The results of GPCR experiments showed that Corydalis Rhizoma could activate 5-HT1 A, OPRM1, ADRB2 receptors and block D_2 receptor. Protopine showed antagonism on D_2 and M_2 receptors, tetrahydropalmatine could agitate ADRB2 receptor and antagonize D_2 and TP receptors, while palmatine showed no significant biological activity on the 6 GPCRs. In conclusion, Corydalis Rhizoma may exert biological activity by multi-components acting on multi-targets.