沙葱总黄酮对过氧化氢诱导的红细胞氧化损伤的保护作用
|
摘要
本试验旨在研究沙葱总黄酮在体外条件下对过氧化氢(H2O2)诱导的红细胞氧化损伤的保护作用。在筛选建立红细胞氧化损伤模型时所需的最适H2O2浓度和作用时间及沙葱总黄酮安全浓度范围的基础上,将试验分为空白对照组、H2O2损伤组、不同浓度沙葱总黄酮组(沙葱总黄酮在细胞悬液中的终浓度分别为30、130、260、390和520μg/mL),每组9个重复,评价沙葱总黄酮对红细胞溶血率、丙二醛(MDA)含量及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)活性的影响。结果表明,沙葱总黄酮能显著抑制H2O2损伤引起的红细胞溶血(P<0.05),且其作用呈剂量依赖关系。与H2O2损伤组相比,130、260和520μg/mL沙葱总黄酮能显著降低红细胞中M DA的含量(P<0.05),并提高红细胞中SOD、CAT和GSH-Px的活性(P<0.05)。由此可见,沙葱总黄酮通过抑制H2O2氧化损伤引起的溶血、降低过氧化产物的产生及提高抗氧化酶活性,从而保护红细胞免受氧化应激引起的损伤。
The aim of this study was to investigate the protective effect of total flavonoids from Allium mongolicum Regel on oxidative damage of erythrocytes induced by hydrogen peroxide(H_2O_2) in vitro.Based on the optimal H_2O_2 concentration and induction time required for the establishment of erythrocytes oxidative damage model and the safe concentration range of total flavonoids from Allium mongolicum Regel,the present experiment were divided into control group,H_2O_2 damage group and different concentrations of total flavonoids from Allium mongolicum Regel groups(the final concentrations of total flavonoids from Allium mongolicum Regel in cell suspension were 30,130,260,390 and 520 μg/mL,respectively),each group had 9 replicates.Subsequently,the effects of total flavonoids from Allium mongolicum Regel on hemolysis rate of erythrocytes,melondialdehyde(MDA) content,activities of superoxide dismutase(SOD),catalase(CAT) and glutathione peroxidase(GSH-Px) were evaluated.The results showed that total flavonoids from Allium mongolicum Regel could significantly inhibit the hemolysis of erythrocytes induced by H_2O_2 oxidative stress(P<0.05),which the effect was dose dependent.Compared with the H_2O_2 damage group,130,260 and 520 μg/mL total flavonoids from Allium mongolicum Regel significantly reduced the erythrocytes MDA content(P<0.05),and significantly increased the activities of SOD,CAT and GSH-Px in erythrocytes(P<0.05).To sum up,total flavonoids from Allium mongolicum Regel can inhibit the hemolysis induced by H_2O_2 oxidative stress,reduce the production of peroxide products and increase the activities of antioxidant enzymes,thereby protect erythrocytes from damages of oxidative stress.
The aim of this study was to investigate the protective effect of total flavonoids from Allium mongolicum Regel on oxidative damage of erythrocytes induced by hydrogen peroxide(H_2O_2) in vitro.Based on the optimal H_2O_2 concentration and induction time required for the establishment of erythrocytes oxidative damage model and the safe concentration range of total flavonoids from Allium mongolicum Regel,the present experiment were divided into control group,H_2O_2 damage group and different concentrations of total flavonoids from Allium mongolicum Regel groups(the final concentrations of total flavonoids from Allium mongolicum Regel in cell suspension were 30,130,260,390 and 520 μg/mL,respectively),each group had 9 replicates.Subsequently,the effects of total flavonoids from Allium mongolicum Regel on hemolysis rate of erythrocytes,melondialdehyde(MDA) content,activities of superoxide dismutase(SOD),catalase(CAT) and glutathione peroxidase(GSH-Px) were evaluated.The results showed that total flavonoids from Allium mongolicum Regel could significantly inhibit the hemolysis of erythrocytes induced by H_2O_2 oxidative stress(P<0.05),which the effect was dose dependent.Compared with the H_2O_2 damage group,130,260 and 520 μg/mL total flavonoids from Allium mongolicum Regel significantly reduced the erythrocytes MDA content(P<0.05),and significantly increased the activities of SOD,CAT and GSH-Px in erythrocytes(P<0.05).To sum up,total flavonoids from Allium mongolicum Regel can inhibit the hemolysis induced by H_2O_2 oxidative stress,reduce the production of peroxide products and increase the activities of antioxidant enzymes,thereby protect erythrocytes from damages of oxidative stress.
引文
[1]包颖.内蒙古葱属植物的地理分布[J].内蒙古师大学报(自然科学版),2000,29(2):130-134.
[2]燕玲,王六英,李红.内蒙古10种葱属植物染色体数目及核型分析[J].中国草地,1999(5):73-75.
[3]李亚蕾,罗瑞明.沙葱提取物抗菌效果的研究[J].安徽农业科学,2008,36(33):14596-14597,14618.
[4]凌关庭.天然抗氧化剂及其消除氧自由基的进展[J].食品工业,2000(3):19-22.
[5]缪亚娟.沙葱异黄酮含量测定及其对机体抗氧化与非特异性免疫影响的研究[D].硕士学位论文.呼和浩特:内蒙古农业大学,2009.
[6]BARRECA D,LAGANG,TELLONE E,et al.Influences of flavonoids on erythrocyte membrane and metabolic implication through anionic exchange modulation[J].Journal of Membrane Biology,2009,230(3):163-171.
[7]张波.杨梅花色苷对胰岛细胞氧化应激损伤的保护作用及其机制探讨[D].博士学位论文.杭州:浙江大学,2010.
[8]SADRZADEH S M,GRAF E,PANTER S S,et al.Hemoglobin.A biologic fenton reagent[J].Journal of Biological Chemistry,1984,259(23):14354-14356.
[9]HONZEL D,CARTER S G,REDMAN K A,et al.Comparison of chemical and cell-based antioxidant methods for evaluation of foods and natural products:generating multifaceted data by parallel testing using erythrocytes and polymorphonuclear cells[J].Journal of Agricultural and Food Chemistry,2008,56(18):8319-8325.
[10]BUEHLER P W,ALAYASH A I.Redox biology of blood revisited:the role of red blood cells in maintaining circulatory reductive capacity[J].Antioxidants&Redox Signaling,2005,7(11/12):1755-1760.
[11]何佳易.红细胞氧化模型评价抗氧化活性的方法学研究[D].硕士学位论文.扬州:扬州大学,2013.
[12]丁航,刘慧明,梁统,等.侧柏叶中黄酮类化合物对H2O2诱导的人红细胞氧化作用的影响[J].实用临床医学,2003,4(3):23-24.
[13]GIRISH T K,VASUDEVARAJU P,PRASADA RAOU J S.Protection of DNA and erythrocytes from free radical induced oxidative damage by black gram(Vigna mungo L.)husk extract[J].Food and Chemical Toxicology,2012,50(5):1690-1696.
[14]赵磊,郝添阳,王旋,等.蛋白水解物对DNA和红细胞氧化损伤的保护作用[J].中国食品学报,2016,16(8):7-15.
[15]皮建辉,胡朝暾,郑晴,等.金樱子多糖体外抗脂质过氧化和红细胞溶血作用研究[J].怀化学院学报,2012,30(5):11-15.
[16]马兰萍,刘在群,周波,等.绿茶多酚对自由基诱导的红细胞氧化性溶血的抑制作用[J].科学通报,2000,45(12):1271-1275.
[17]赵春艳.沙葱中黄酮类化合物的分离纯化、结构鉴定及其对小鼠免疫抗氧化机能影响的研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2008.
[18]木其尔.沙葱黄酮对肉羊机体抗氧化和免疫功能的影响及其机理研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2016.
[19]萨茹丽.沙葱黄酮提取工艺优化、结构鉴定及其相关生物活性研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2014.
[20]刘兵,苑博,宁天一,等.沙棘总黄酮对过氧化氢损伤人血管内皮细胞的保护作用[J].中华中医药学刊,2017,35(8):2158-2160.
[21]FEDERICO A,CARDAIOLI E,DA POZZO P,et al.Mitochondria,oxidative stress and neurodegeneration[J].Journal of the Neurological Sciences,2012,322(1/2):254-262.
[22]RTH E.Oxygen free radicals and their clinical implications[J].Acta Chirurgica Hungarica,1997,36(1/2/3/4):302-305.
[23]NATHAN C.Nitric oxide as a secretory product of mammalian cells[J].The FASEB Journal,1992,6(12):3051-3064.
[24]SALUCCI S,BURATTINI S,BATTISTELLI M,et al.Ultraviolet B(UVB)irradiation-induced apoptosis in various cell lineages in vitro[J].International Journal of Molecular Sciences,2012,14(1):532-546.
[25]张玲莉,邱振鹏.黄芩提取物对红细胞抗氧化作用研究[J].中国药师,2011,14(11):1572-1575.
[26]ARBOS K A,ClARO L M,BORGES L,et al.Human erythrocytes as a system for evaluating the antioxidant capacity of vegetable extracts[J].Nutrition Research,2008,28(7):457-463.
[27]蒙萍,马慧萍,王宁,等.大苞雪莲总黄酮体外抗氧化活性[J].中国实验方剂学杂志,2015,21(17):92-95.
[28]YAMAKOSHI J,SANO A,TOKUTAKE S,et al.Oral intake of proanthocyanid in-rich extract from grape seeds improves chloasma[J].Phytotherapy Research,2004,18(11):895-899.
[29]TEBIB K,ROUANET J M,BESANON P.Antioxidant effects of dietary polymeric grape seed tannins in tissues of rats fed a high cholesterol-vitamin E-deficient diet[J].Food Chemistry,1997,59(1):135-141.
[30]王丹丹.三种植物提取物对H2O2诱导PC12细胞氧化损伤的保护作用研究[D].硕士学位论文.济南:山东师范大学,2016.
[2]燕玲,王六英,李红.内蒙古10种葱属植物染色体数目及核型分析[J].中国草地,1999(5):73-75.
[3]李亚蕾,罗瑞明.沙葱提取物抗菌效果的研究[J].安徽农业科学,2008,36(33):14596-14597,14618.
[4]凌关庭.天然抗氧化剂及其消除氧自由基的进展[J].食品工业,2000(3):19-22.
[5]缪亚娟.沙葱异黄酮含量测定及其对机体抗氧化与非特异性免疫影响的研究[D].硕士学位论文.呼和浩特:内蒙古农业大学,2009.
[6]BARRECA D,LAGANG,TELLONE E,et al.Influences of flavonoids on erythrocyte membrane and metabolic implication through anionic exchange modulation[J].Journal of Membrane Biology,2009,230(3):163-171.
[7]张波.杨梅花色苷对胰岛细胞氧化应激损伤的保护作用及其机制探讨[D].博士学位论文.杭州:浙江大学,2010.
[8]SADRZADEH S M,GRAF E,PANTER S S,et al.Hemoglobin.A biologic fenton reagent[J].Journal of Biological Chemistry,1984,259(23):14354-14356.
[9]HONZEL D,CARTER S G,REDMAN K A,et al.Comparison of chemical and cell-based antioxidant methods for evaluation of foods and natural products:generating multifaceted data by parallel testing using erythrocytes and polymorphonuclear cells[J].Journal of Agricultural and Food Chemistry,2008,56(18):8319-8325.
[10]BUEHLER P W,ALAYASH A I.Redox biology of blood revisited:the role of red blood cells in maintaining circulatory reductive capacity[J].Antioxidants&Redox Signaling,2005,7(11/12):1755-1760.
[11]何佳易.红细胞氧化模型评价抗氧化活性的方法学研究[D].硕士学位论文.扬州:扬州大学,2013.
[12]丁航,刘慧明,梁统,等.侧柏叶中黄酮类化合物对H2O2诱导的人红细胞氧化作用的影响[J].实用临床医学,2003,4(3):23-24.
[13]GIRISH T K,VASUDEVARAJU P,PRASADA RAOU J S.Protection of DNA and erythrocytes from free radical induced oxidative damage by black gram(Vigna mungo L.)husk extract[J].Food and Chemical Toxicology,2012,50(5):1690-1696.
[14]赵磊,郝添阳,王旋,等.蛋白水解物对DNA和红细胞氧化损伤的保护作用[J].中国食品学报,2016,16(8):7-15.
[15]皮建辉,胡朝暾,郑晴,等.金樱子多糖体外抗脂质过氧化和红细胞溶血作用研究[J].怀化学院学报,2012,30(5):11-15.
[16]马兰萍,刘在群,周波,等.绿茶多酚对自由基诱导的红细胞氧化性溶血的抑制作用[J].科学通报,2000,45(12):1271-1275.
[17]赵春艳.沙葱中黄酮类化合物的分离纯化、结构鉴定及其对小鼠免疫抗氧化机能影响的研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2008.
[18]木其尔.沙葱黄酮对肉羊机体抗氧化和免疫功能的影响及其机理研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2016.
[19]萨茹丽.沙葱黄酮提取工艺优化、结构鉴定及其相关生物活性研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2014.
[20]刘兵,苑博,宁天一,等.沙棘总黄酮对过氧化氢损伤人血管内皮细胞的保护作用[J].中华中医药学刊,2017,35(8):2158-2160.
[21]FEDERICO A,CARDAIOLI E,DA POZZO P,et al.Mitochondria,oxidative stress and neurodegeneration[J].Journal of the Neurological Sciences,2012,322(1/2):254-262.
[22]RTH E.Oxygen free radicals and their clinical implications[J].Acta Chirurgica Hungarica,1997,36(1/2/3/4):302-305.
[23]NATHAN C.Nitric oxide as a secretory product of mammalian cells[J].The FASEB Journal,1992,6(12):3051-3064.
[24]SALUCCI S,BURATTINI S,BATTISTELLI M,et al.Ultraviolet B(UVB)irradiation-induced apoptosis in various cell lineages in vitro[J].International Journal of Molecular Sciences,2012,14(1):532-546.
[25]张玲莉,邱振鹏.黄芩提取物对红细胞抗氧化作用研究[J].中国药师,2011,14(11):1572-1575.
[26]ARBOS K A,ClARO L M,BORGES L,et al.Human erythrocytes as a system for evaluating the antioxidant capacity of vegetable extracts[J].Nutrition Research,2008,28(7):457-463.
[27]蒙萍,马慧萍,王宁,等.大苞雪莲总黄酮体外抗氧化活性[J].中国实验方剂学杂志,2015,21(17):92-95.
[28]YAMAKOSHI J,SANO A,TOKUTAKE S,et al.Oral intake of proanthocyanid in-rich extract from grape seeds improves chloasma[J].Phytotherapy Research,2004,18(11):895-899.
[29]TEBIB K,ROUANET J M,BESANON P.Antioxidant effects of dietary polymeric grape seed tannins in tissues of rats fed a high cholesterol-vitamin E-deficient diet[J].Food Chemistry,1997,59(1):135-141.
[30]王丹丹.三种植物提取物对H2O2诱导PC12细胞氧化损伤的保护作用研究[D].硕士学位论文.济南:山东师范大学,2016.