汉源花椒总黄酮提取物的抗氧化及抗Hela肿瘤细胞增殖活性研究

The Study of Anti-oxidative and Anti-proliferative Activities of Total Flavonoids Extract from Hanyuan Zanthoxylum bungeanum

本研究采用超声波辅助溶剂提取、大孔树脂纯化法制备了汉源花椒总黄酮提取物,研究其抗氧化活性及对人宫颈癌Hela细胞的抗增殖活性。采用1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)法、水杨酸法、磷钼络合物法和三价铁还原法研究提取物的体外抗氧化活性,采用CCK8法、PI染色法和EGFP-Annexin V/PI染色法研究提取物对Hela细胞的抗增殖活性影响。结果表明:总黄酮提取得率为2.19%;当提取物质量浓度为125、500μg/mL时,对DPPH自由基和羟自由基(·OH)的清除率分别为84.70%和83.61%,IC50值分别为65.61、194.40μg/mL;当提取物质量浓度为400和300μg/mL时还原力与总抗氧化能力最强,吸光度分别为0.70和0.48。在细胞实验中,用800μg/mL的提取物处理12、24、48 h后的Hela细胞生存率分别为27.15%、10.43%和30.63%,与相同处理时间的对照组相比,生存率极显著下降(P<0.001),这可能与该浓度提取物处理24 h的细胞发生G2/M期阻滞和细胞凋亡率升高有关。因此,汉源花椒总黄酮提取物具有一定抗氧化和对Hela细胞的抗增殖活性,该研究结果可为进一步开发利用花椒资源提供理论支持。

Total flavonoids of Hanyuan Zanthoxylum bungeanum were prepared by ultrasonic-assisted solvent extraction and macroporous resins purification in this study,and the anti-oxidative activity in vitro as well as anti-proliferative activity on Hela cells was studied.The anti-oxidative activity in vitro was detected by the scavenging rates of DPPH·and hydroxyl radicals,the formation of phosphorus molybdenum complex and the reducing ability of Fe^3+.The anti-proliferative activities on Hela cells were detected by CCK-8 as well as the fluorescent staining methods of PI and EGFP-Annexin V/PI.The results showed that the yield of total flavonoids was 2.19%.The scavenging rates of DPPH and hydroxyl radicals were 84.70% and 83.61%respectively at the concentrations of 125 and 500μg/mL,and the IC50 values were 65.61 and 194.40μg/mL respectively.The reducing power and total anti-oxidative capacity were the strongest at the concentrations of 400 and 300μg/mL,and the absorbance of reducing power and total antioxidant capacity were 0.70 and 0.48,respectively.In the study of cells,the viabilities of Hela cells were 27.15%,10.43% and 30.63% respectively after 12,24,and 48 h treated by the concentration of 800μg/mL,and decreased significantly(P<0.001)compared to the control at the same time,which could be due to cell cycle arrest induced in G2/M phase and the cell apoptosis rate increased at this mass concentration at 24 h.Therefore,the total flavonoids extract from Hanyuan Zanthoxylum bungeanum had the anti-oxidative activity in vitro and antiproliferative activity on Hela cells,which could be theoretical support for further using Hanyuan Zanthoxylum bungeanum as a source of some functional products.