人参-黄芪药对不同配比化学成分分析及抗氧化活性研究

Analysis of chemical constituents and antioxidant activity of different ratios of ginseng and Astragalus

目的:比较不同配比人参-黄芪药对中指标性成分差异,探究其抗氧化活性。方法:制备人参、黄芪提取物及不同配伍比例(1∶1,1∶2,1∶4,m/m)的人参-黄芪药对提取物,采用1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)和2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸+[2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonate+,ABTS+]和等化学法检测人参-黄芪药对清除自由基能力;建立H_(2)O_(2)诱导的HepG2细胞氧化损伤模型,通过荧光探针法与ELISA法分析人参-黄芪药对对HepG2细胞模型中氧化因子分泌水平的影响;建立D-半乳糖诱导小鼠衰老模型分析人参-黄芪药对不同配比的体内抗氧化活性;采用紫外分光光度法和HPLC-ELSD法,分析比较各提取物中活性成分(总酚酸、总皂苷、总多糖、总黄酮、总蛋白)及7种主要化学成分的含量差异。结果:人参-黄芪药对不同配比提取物,均显示出较好的抗氧化活性,人参-黄芪(1∶2)药对具有较强的清除自由基能力。体外细胞模型实验中,人参-黄芪药对给药后可显著提高细胞活力、超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)含量,降低丙二醛(malondialdehyde,MDA)浓度,其中,人参-黄芪(1∶2)给药浓度为400μg·mL^(-1)时,细胞活力最高可达(92.88±2.81)%。体内实验表明,人参-黄芪(1∶2)可以增强模型小鼠血液和肝脏中超氧化物歧化酶和谷胱甘肽过氧化物酶含量,降低丙二醛浓度。人参-黄芪药对活性成分分析结果表明,人参-黄芪(1∶2)配比时总黄酮和总皂苷含量最高。结论:与单药材提取物相比,人参-黄芪药对不同配比提取物均具有良好的清除自由基能力及体内体外抗氧化作用,其中人参-黄芪(1∶2)药对的抗氧化活性最好。

Objective:To compare the differences at index components in different ratios of ginseng-Astragalus membranaceus pairs and to investigate their antioxidant activities.Methods:Extracts of ginseng and astragalus and extracts of ginseng-astragalus pairs with different pairing ratios(1∶1,1∶2,1∶4,m/m)were prepared,and the scavenging ability of ginseng-astragalus pairs was detected by chemical methods such as 1,1-diphenyl-2-trinitrophenylhydrazine(DPPH)and 2,2'-biazobis-3-ethylbenzothiazoline-6-sulfonic acid^(+)(ABTS^(+));a model of H_(2)O_(2)-induced oxidative damage in HepG2 cells was established.H_(2)O_(2)-induced oxidative damage model of HepG2 cells,and analyzed the effect of ginseng-Astragalus medicine on the secretion level of oxidative factors in the HepG2 cell model by fluorescent probe method and ELISA;established D-Galactose-induced mouse aging model to analyze the in vivo antioxidant activity of ginseng-Astragalus medicine on different ratios;and analyzed and compared the active ingredients(total and total)of the extracts by using the UV spectrophotometric method and HPLC-ELSD method.The differences in the contents of active ingredients(total phenolic acids,total saponins,total polysaccharides,total flavonoids,total proteins)and seven major chemical components in the extracts were analyzed and compared using UV spectrophotometry and HPLC-ELSD.Results:The ginseng-Astragalus medicinal pairs with different ratios of extracts showed better antioxidant activities,and the ginseng-Astragalus(1∶2)medicinal pairs possessed stronger scavenging ability of free radicals.In the in vitro cell modeling assay,the ginseng-Astragalus drug pair administration significantly increased the cell viability,superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)contents,and decreased the malondialdehyde(MDA)concentration,of which the highest cell viability of ginseng-Astragalus(1∶2)could be reached at the administration concentration of 400μg·mL^(-1)(92.88±2.81)%.In vivo experiments showed that ginseng-Astragalus membranaceus(1∶2)enhanced superoxide dismutase and glutathione peroxidase levels and decreased malondialdehyde concentration in blood and liver of model mice.The results of Ginseng-Astragalus drug-pair active ingredient analysis showed that the highest total flavonoids and total saponins were found in the Ginseng-Astragalus(1∶2)ratio.Conclusion:Compared with single herb extracts,the different ratios of ginseng-Astragalus medicinal pairs had good free radical scavenging ability and antioxidant effects in vivo and in vitro,with the best antioxidant activity of ginseng-Astragalus(1∶2)medicinal pairs.