广东凉茶提取物对兔红细胞膜氧化损伤保护作用研究
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摘要
目的从膜成分、结构和功能角度观察广东凉茶提取物对兔红细胞膜氧化损伤的保护作用。方法实验设对照组、H2O2组(1 000μmol/L)、0.1 mg组(0.1 mg/mL凉茶+H2O2)、0.2 mg组(0.2 mg/mL凉茶+H2O2)、0.4 mg组(0.4 mg/mL凉茶+H2O2),参照Dodge法制备兔红细胞膜,观察各组对丙二醛(MDA)含量、蛋白羰基含量、膜封闭能力、细胞膜ATP酶活力、谷胱甘肽过氧化物酶(GPX)活力、过氧化氢酶(CAT)活力和细胞膜蛋白成分的影响。结果与对照组相比,H2O2组蛋白羰基含量、GPX酶活力升高,细胞膜封闭率、CAT酶活力、Ca2+Mg2+?ATP、Mg2+?ATP、Na+K+?ATP酶活力降低(均P<0.01)。与H2O2组比,广东凉茶提取物0.1 mg组、0.2 mg组、0.4 mg组的MDA和DNPH含量均降低,细胞膜封闭率、Ca2+Mg2+?ATP、Mg2+?ATP、Na+K+?ATP酶活力、GPX酶活力、CAT酶活力升高,差异均有统计学意义(均P<0.05)。结论广东凉茶提取物可抑制H2O2引起的红细胞膜成分、结构和功能的损伤作用。
Objective To observe the protective effect of Guangdong herbal tea extract on oxidative damage of rabbit erythrocyte membrane from the perspective of membrane composition,structure and function. Methods The experiment consisted of a control group,H2 O2 group(1 000 μmol/L),0.1 mg group(0.1 mg/ml herbal tea +1 000 μmol/L H202),0.2 mg group(0.2 mg/mL herbal tea +1 000 μmol/L H202),and 0.4 mg group(0.4 mg/mL herbal tea +1 000 μmol/L H202). Rabbit red blood cell membrane was prepared by Dodge method. The effects of malondialdehyde(MDA),the carbonyl content of protein,membrane blocking ability,ATPase activity,glutathione peroxidase(GPX),catalase(CAT)activity,and membrane protein composition on each group were observed. Results Compared with the control group,the carbonyl content of protein and GPX activity were increased,while cell membrane blocking ability,activity of CAT,Ca2+Mg2+ATPase,Mg2+ATPase,and Na+K+ATPase in H2 O2 group were decreased(P < 0.01 for all). Compared with the H2 O2 group,contents of MDA and DNPH decreased,while the cell membrane blocking rate,and activities of Ca2+Mg2+ATPase,Mg2+ATPase,Na+K+ATPase,GPX,and CAT increased in the three herbal tea groups(P < 0.05 for all). Conclusion Guangdong herbal tea extract could inhibit the damage of erythrocyte membrane composition,structure and function caused by hydrogen peroxide.
Objective To observe the protective effect of Guangdong herbal tea extract on oxidative damage of rabbit erythrocyte membrane from the perspective of membrane composition,structure and function. Methods The experiment consisted of a control group,H2 O2 group(1 000 μmol/L),0.1 mg group(0.1 mg/ml herbal tea +1 000 μmol/L H202),0.2 mg group(0.2 mg/mL herbal tea +1 000 μmol/L H202),and 0.4 mg group(0.4 mg/mL herbal tea +1 000 μmol/L H202). Rabbit red blood cell membrane was prepared by Dodge method. The effects of malondialdehyde(MDA),the carbonyl content of protein,membrane blocking ability,ATPase activity,glutathione peroxidase(GPX),catalase(CAT)activity,and membrane protein composition on each group were observed. Results Compared with the control group,the carbonyl content of protein and GPX activity were increased,while cell membrane blocking ability,activity of CAT,Ca2+Mg2+ATPase,Mg2+ATPase,and Na+K+ATPase in H2 O2 group were decreased(P < 0.01 for all). Compared with the H2 O2 group,contents of MDA and DNPH decreased,while the cell membrane blocking rate,and activities of Ca2+Mg2+ATPase,Mg2+ATPase,Na+K+ATPase,GPX,and CAT increased in the three herbal tea groups(P < 0.05 for all). Conclusion Guangdong herbal tea extract could inhibit the damage of erythrocyte membrane composition,structure and function caused by hydrogen peroxide.
引文
[1]王兰,张晓文,赵广才,等.中草药茶饮料饮品凉茶的功效及安全性研究评述[J].中医学报,2010,25(5):914-916.
[2]何蓉蓉,姚新生,栗原博.广东凉茶的“泻火”作用与物质基础研究[J].世界科学技术(中医药现代化),2009,11(6):834-839.
[3]谢果,李善兵,何蓉蓉,等.王老吉凉茶对拘束应激负荷小鼠脾脏CTL活性的影响[J].中药新药与临床药理,2010,21(3):223-226.
[4]宝丽,姚新生,栗原博,等.广东凉茶颗粒对拘束负荷诱发小鼠应激性肝损伤的保护作用[J].中国中药杂志,2008,23(6):664-668.
[5] Dodge JT,Mitchell C,Hanahan DJ. The preparation and chemical characteristics of hemoglobinfree ghosts of human erythrocyte[J]. Arch Biochim Biophys,1963,100(1):119–130.
[6]刘彬,齐云,李蒙,等.分光光度法与荧光法测定肝组织丙二醛含量的比较研究[J].中国药理学通报,2010,26(12):1674-1677.
[7]段丽菊,刘英帅,朱燕,等. DNPH比色法:一种简单的蛋白质羰基含量测定方法[J].毒理学杂志,2005,19(4):320-322.
[8]王晓玲,秦德安.橙皮苷对羟自由基引发红细胞膜损伤的影响[J].中国药学杂志,1998,33(2):83-85.
[9]姜维华.人红细胞膜葡萄糖转运蛋白特性及其相互作用蛋白质的研究[D].上海:复旦大学,2005.
[10] Shan YP,Wang HD. The structure and function of cell membranes examined by atomic force microscopy and singlemolecule force spectroscopy[J]. Chem Soc Rev,2015,44(11):3617-3638.
[11] Revin VV,Filatova SM,Syusin IV,et al. Study of correlation between state and composition of lipid phase and change in erythrocytes structure under induction of oxidative processes[J]. Int J Hematol,2015,101(5):487-496.
[12] Tsikas D. Assessment of lipid peroxidation by measuring malondialdehyde(MDA)and relatives in biological samples:analytical and biological challenges[J]. Anal Biochem,2017,524:13-30.
[13] Kumar N,Maurya PK,Kant R,et al.(-)Epicatechin in vitro ameliorates erythrocyte protein carbonyl content in hypertensive patients:comparison with Lascorbic acid[J]. Arch Physiol Biochem,2016,122(3):155-160.
[14] Alberts B(美).细胞的分子生物学[M].张新跃,钱万强,译.北京:科学出版社,2008:651-652.
[15] Bukowska B,Sicinska P,Pajak A,et al. Oxidative stress and damage to erythrocytes in patients with chronic obstructive pulmonary diseasechanges in ATPase and acetylcholinesterase activity[J]. Biochem Cell Biol,2015,93(6):574-580.
[16] Rocha S,Gomes D,Lima M,et al. Peroxiredoxin 2,glutathione peroxidase,and catalase in the cytosol and membrane of erythrocytes under H2O2induced oxidative stress[J]. Free Radical Research,2015,49(8):990-1003.
[17] Sani M,Sebai H,GhanemBoughanmi N,et al. Circadian(about24hour)variation in malondialdehyde content and catalase activityofmouseerythrocytes[J].ReodxReport,2015,20(1):26-32.
[18]王辉,周立,刘刚.莲心碱对氧自由基引发红细胞膜损伤的保护作用[J].中国药理学通报,2006,22(9):1088-1091.
[19] Lubos E,Loscalzo J,Handy DE. Glutathione peroxidase1 in health and disease:from molecular mechanisms to therapeutic opportunities[J]. Antioxid Redox Signal,2011,15(7):1957-1997.
[2]何蓉蓉,姚新生,栗原博.广东凉茶的“泻火”作用与物质基础研究[J].世界科学技术(中医药现代化),2009,11(6):834-839.
[3]谢果,李善兵,何蓉蓉,等.王老吉凉茶对拘束应激负荷小鼠脾脏CTL活性的影响[J].中药新药与临床药理,2010,21(3):223-226.
[4]宝丽,姚新生,栗原博,等.广东凉茶颗粒对拘束负荷诱发小鼠应激性肝损伤的保护作用[J].中国中药杂志,2008,23(6):664-668.
[5] Dodge JT,Mitchell C,Hanahan DJ. The preparation and chemical characteristics of hemoglobinfree ghosts of human erythrocyte[J]. Arch Biochim Biophys,1963,100(1):119–130.
[6]刘彬,齐云,李蒙,等.分光光度法与荧光法测定肝组织丙二醛含量的比较研究[J].中国药理学通报,2010,26(12):1674-1677.
[7]段丽菊,刘英帅,朱燕,等. DNPH比色法:一种简单的蛋白质羰基含量测定方法[J].毒理学杂志,2005,19(4):320-322.
[8]王晓玲,秦德安.橙皮苷对羟自由基引发红细胞膜损伤的影响[J].中国药学杂志,1998,33(2):83-85.
[9]姜维华.人红细胞膜葡萄糖转运蛋白特性及其相互作用蛋白质的研究[D].上海:复旦大学,2005.
[10] Shan YP,Wang HD. The structure and function of cell membranes examined by atomic force microscopy and singlemolecule force spectroscopy[J]. Chem Soc Rev,2015,44(11):3617-3638.
[11] Revin VV,Filatova SM,Syusin IV,et al. Study of correlation between state and composition of lipid phase and change in erythrocytes structure under induction of oxidative processes[J]. Int J Hematol,2015,101(5):487-496.
[12] Tsikas D. Assessment of lipid peroxidation by measuring malondialdehyde(MDA)and relatives in biological samples:analytical and biological challenges[J]. Anal Biochem,2017,524:13-30.
[13] Kumar N,Maurya PK,Kant R,et al.(-)Epicatechin in vitro ameliorates erythrocyte protein carbonyl content in hypertensive patients:comparison with Lascorbic acid[J]. Arch Physiol Biochem,2016,122(3):155-160.
[14] Alberts B(美).细胞的分子生物学[M].张新跃,钱万强,译.北京:科学出版社,2008:651-652.
[15] Bukowska B,Sicinska P,Pajak A,et al. Oxidative stress and damage to erythrocytes in patients with chronic obstructive pulmonary diseasechanges in ATPase and acetylcholinesterase activity[J]. Biochem Cell Biol,2015,93(6):574-580.
[16] Rocha S,Gomes D,Lima M,et al. Peroxiredoxin 2,glutathione peroxidase,and catalase in the cytosol and membrane of erythrocytes under H2O2induced oxidative stress[J]. Free Radical Research,2015,49(8):990-1003.
[17] Sani M,Sebai H,GhanemBoughanmi N,et al. Circadian(about24hour)variation in malondialdehyde content and catalase activityofmouseerythrocytes[J].ReodxReport,2015,20(1):26-32.
[18]王辉,周立,刘刚.莲心碱对氧自由基引发红细胞膜损伤的保护作用[J].中国药理学通报,2006,22(9):1088-1091.
[19] Lubos E,Loscalzo J,Handy DE. Glutathione peroxidase1 in health and disease:from molecular mechanisms to therapeutic opportunities[J]. Antioxid Redox Signal,2011,15(7):1957-1997.