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目的了解5种黄连生物碱(巴马亭、黄连碱、表小檗碱、药根碱和小檗碱)与生物大分子蛋白质和DNA的作用关系及其差异。方法将黄连生物碱巴马亭、黄连碱、表小檗碱、药根碱和小檗碱均配成浓度分别为0、1.95×10-6、3.9×10-6、7.8×10-6、1.56×10-5和3.12×10-5 mol/L的PBS稀释液后,分别量取1.5mL与0.5mL牛血清白蛋白(BSA,1.10×10-5 mol/L)溶液作用,采用F-4500型荧光分光光度计,在280nm激发波长条件下扫描300~500nm范围的荧光发射光谱。将浓度均为0、1.95×10-6、3.9×10-6、7.8×10-6和1.56×10-5 mol/L的5种黄连生物碱PBS稀释液各量取1.5mL与0.5mL百分浓度为0.1%的质粒DNA作用后,在368nm激发波长条件下扫描480~650nm范围的荧光发射光谱。结果 5种黄连生物碱对BSA荧光均有很强的淬灭作用,各生物碱之间的荧光淬灭作用差异不大。5种黄连生物碱与质粒DNA作用后,均能增强发射荧光,且各生物碱之间的荧光增强作用差异很大,荧光增强作用由大到小依次为表小檗碱、黄连碱、巴马亭、小檗碱和药根碱,其中,表小檗碱和黄连碱增强作用远大于其余3种黄连生物碱;5种黄连生物碱的最强荧光作用浓度均为7.8×10-6 mol/L。结论 5种黄连生物碱与蛋白和DNA均存在很强的相互作用。5种黄连生物碱与蛋白之间的作用差异较小,与DNA之间的作用差异较大,提示黄连生物碱与DNA的作用强度与其分子结构有关,该类分子中9,10-亚甲二氧基和2,3-二甲氧基结构有利于与DNA作用后的荧光增强作用。
OBJECTIVE To investigate the effects and relationships of the five Coptis alkaloids (parmating, berberine, berberine, jatrorrhizine and berberine) on the biological macromolecules and DNA. Methods Coptis chinensis alkaloids, palmatine, berberine, jatrorrhizine and berberine were all formulated into concentrations of 0, 1.95 × 10-6, 3.9 × 10-6, 7.8 × 10-6, 1.56 × 10-5 and 3.12 × 10-5 mol / L PBS, 1.5mL and 0.5mL bovine serum albumin (BSA, 1.10 × 10-5 mol / L) solution were measured respectively. F- 4500 fluorescence spectrophotometer, under the excitation wavelength of 280nm scanning fluorescence emission spectrum in the range of 300 ~ 500nm. Five concentrations of Coptis alkaloid PBS dilutions with concentrations of 0, 1.95 × 10-6, 3.9 × 10-6, 7.8 × 10-6 and 1.56 × 10-5 mol / L were respectively taken as 1.5mL and 0.5mL Subsequent to the action of plasmid DNA at a concentration of 0.1%, a fluorescence emission spectrum in the range of 480-650 nm was scanned under an excitation wavelength of 368 nm. Results The five kinds of Coptis alkaloids had a strong quenching effect on the BSA fluorescence and the difference of fluorescence quenching between alkaloids was not significant. After the action of 5 kinds of Coptis alkaloids and plasmid DNA, the emission fluorescence was enhanced, and the fluorescence enhancement effect between each alkaloid was quite different. The enhancement of fluorescence was descending order of berberine, coptisine, Paeoniflorin, berberine and jatrorrhizine. Among them, the enhancing effect of berberine and coptisine was much greater than that of the other three berberine alkaloids. The strongest fluorescence concentrations of the five kinds of berberine alkaloids were 7.8 × 10-6 mol / L. Conclusion Five kinds of Coptis alkaloids have strong interaction with protein and DNA. The differences between the five alkaloids of Coptis chinensis and the protein are small and the differences between the two alkaloids and DNA are quite different. It suggests that the interaction between Coptis alkaloids and DNA is related to its molecular structure. Among these molecules, 9,10-methylenebis The oxy and 2,3-dimethoxy structures favor fluorescence enhancement after DNA interaction.