苦荞黄酮的抗脂质过氧化和红细胞保护作用研究

Study on Antioxidant and Red Blood Cell Protecting Properties of Tartary Buckwheat Flavonoids

目的:研究苦荞总黄酮的体外抗脂质过氧化和红细胞保护作用,分析其抗氧化的主要成分。方法:将苦荞总黄酮用200～300目硅胶柱层析分离,氯仿-甲醇-水梯度洗脱,紫外检测,得到18个Rf值不同部位;DPPH活性示踪,得到2个活性较强部位(Fr4、Fr9);采用大鼠肝组织脂质过氧化、红细胞溶血模型比较苦荞总黄酮、Fr4、Fr9、槲皮素及芦丁的抗氧化效应;用薄层色谱分析Fr4、Fr9的化学组成。结果:苦荞总黄酮、Fr4、Fr9、槲皮素及芦丁DPPH抑制率(IR)分别为53.13%、66.15%、68.55%、71.99%、63.08%;抑制大鼠肝脏自发性脂质过氧化半抑制浓度(IC50)分别为:27.78、16.05、14.28、8.74和7.4mg/mL;抑制H2O2诱导大鼠肝脂质过氧化IC50分别为:0.37、3.60、0.07、0.07和0.41mg/mL;抑制H2O2诱导大鼠红细胞溶血IC50分别为:13.00、0.48、0.20、0.08和4.10mg/mL。Fr4、Fr9均含有槲皮素,另有3个组分待定。结论:槲皮素是苦荞总黄酮在体外表现抗脂质过氧化和红细胞保护作用主要活性成分之一。

Objective： To study lipid peroxidation inhibiting and red blood cell protecting activities of tartary buckwheat flavonoids, analysis the main antioxidant component. Methods： The total flavonoids of tartary buckwheat flour （TFTBF） was separated by silica column chromatography and 18 fractions were obtained. According to the DPPH scavenging activity assay, Fr4 and Fr9 had the stronger antioxidative effect. The antioxidative activities of TFTBF, Fr4, Fr9, quercetin and rutin were compared each other by rat liver and red blood cell models. The components of Fr4 and Fr9 were analyzed by thin layer chromatography. Results： The inhibition of DPPH free radical of TFTBF, Fr4, Fr9, quercetin and rutin were 53.13%, 66.15%, 68.55%, 71.99% and 63.08% respectively. In rat liver lipid autoperoxidation assay, the IC50 values of the samples mentioned above were 27.78 mg/mL, 16.05 mg/mL, 14.28 mg/mL, 8.74 mg/mL and 7.4 mg/mL respectively. In rat liver lipid peroxidation induced by H202 assay, the IC50 values of the samples were 0.37 mg/mL, 3.60 mg/mL, 0.07 mg/mL, 0.07 mg/mL and 0.41 mg/mL respectively. In rat red blood cell hemolysis induced by H202 assay, the IC50 values of the samples were 13.00 mg/mL, 0.48 mg/mL, 0.20 mg/mL, 0.08 mg/mL and 4.10 mg/mL respectively. Both Fr4 and Fr9 consisted of quercetin and the other three unknown compounds. Conclusion： Quereetin is the one of important antioxidant components in TFTBF.