牡丹花蕊黄酮对H_2O_2诱导RAW264.7巨噬细胞损伤的保护作用
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摘要
目的:研究牡丹花蕊黄酮对H_2O_2诱导损伤的RAW264.7巨噬细胞的保护作用。方法:实验分为空白组、H_2O_2损伤组、VC对照组和牡丹花蕊黄酮低、中、高剂量组。应用H_2O_2建立RAW264.7巨噬细胞氧化损伤模型,噻唑蓝法测定细胞存活率,试剂盒法测定细胞和细胞培养液中谷胱甘肽和丙二醛含量及过氧化氢酶、超氧化物歧化酶、乳酸脱氢酶活力。结果:牡丹花蕊黄酮的质量浓度在10～30μg/mL范围内时,可显著促进RAW264.7巨噬细胞的增殖(P<0.05)。与H2O2损伤组相比,牡丹花蕊黄酮各剂量组能够提高细胞及细胞培养液中总抗氧化能力,谷胱甘肽过氧化物酶、超氧化物酶、过氧化氢酶活力以及还原型谷胱甘肽含量,降低细胞及细胞培养液中丙二醛含量,提高细胞内乳酸脱氢酶的水平。结论:牡丹花蕊黄酮可以促进RAW264.7巨噬细胞的增殖,提高H_2O_2损伤的RAW264.7巨噬细胞的抗氧化能力,且存在剂量依赖效应。
Objective: The aim of this study was to investigate the protective effect of flavonoids from peony stamens(PSF) against hydrogen peroxide(H_2O_2)-induced injury in RAW264.7 cells. Methods: This experiment was divided into blank, model, control and low-, medium-and high-dose PSF groups. RAW264.7 cells were injured by H_2O_2 and the cell viability was measured by methyl thiazolyl tetrazolium assay. The contents of malondialdehyde(MDA) and glutathione(GSH) and the activities of catalase(CAT), superoxide dismutase(SOD) and lactate dehydrogenase(LDH) inside the cells and in the culture medium were determined by commercial kits. Results: The flavonoids at concentrations in the range of 10–30 μg/m L significantly enhanced the proliferation of RAW264.7 cells(P < 0.05). Compared with the model group, the flavonoids at all concentrations could increase total antioxidant capacity(T-AOC), the activities of glutathione peroxidase(GSH-Px), SOD and CAT and GSH content, reduce MDA content in the cultured cells and culture medium and improve LDH activity in the cells. Conclusion: Flavonoids from peony stamens can enhance the proliferation of RAW264.7 cells and protect them against H_2O_2-induced toxicity in a dose-dependent manner.
Objective: The aim of this study was to investigate the protective effect of flavonoids from peony stamens(PSF) against hydrogen peroxide(H_2O_2)-induced injury in RAW264.7 cells. Methods: This experiment was divided into blank, model, control and low-, medium-and high-dose PSF groups. RAW264.7 cells were injured by H_2O_2 and the cell viability was measured by methyl thiazolyl tetrazolium assay. The contents of malondialdehyde(MDA) and glutathione(GSH) and the activities of catalase(CAT), superoxide dismutase(SOD) and lactate dehydrogenase(LDH) inside the cells and in the culture medium were determined by commercial kits. Results: The flavonoids at concentrations in the range of 10–30 μg/m L significantly enhanced the proliferation of RAW264.7 cells(P < 0.05). Compared with the model group, the flavonoids at all concentrations could increase total antioxidant capacity(T-AOC), the activities of glutathione peroxidase(GSH-Px), SOD and CAT and GSH content, reduce MDA content in the cultured cells and culture medium and improve LDH activity in the cells. Conclusion: Flavonoids from peony stamens can enhance the proliferation of RAW264.7 cells and protect them against H_2O_2-induced toxicity in a dose-dependent manner.
引文
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[24]苗明三,陈元朋,吴巍.姜黄素对大鼠血瘀性脑缺血模型血液流变学及脑匀浆LD、LDH和TchE水平的影响[J].中药药理与临床,2010, 26(1):29-31. DOI:10.13412/j.cnki.zyyl.2010.01.009.
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[26]刘思思,李琦,孙立东,等.参莲提取物对LPS诱导的巨噬细胞炎症反应的影响[J].中国实验方剂学杂志, 2017, 23(10):85-91.DOI:10.13422/j.cnki.syfjx.2017100085.
[27]宮甜甜,黄少刚,张玥,等.巨噬细胞的极化及功能调控[J].解剖学报,2017,48(1):106-110.DOI:10.16098/j.issn.0529-1356.2017.01.019.
[28]袁芳.山楂叶总黄酮对H2O2诱导乳鼠心肌细胞氧化应激和细胞凋亡的影响[J].现代中西医结合杂志, 2016, 25(4):368-371; 405.DOI:10.3969/j.issn.1008-8849.2016.04.008.
[29]刘栩岑,李玉洁,王娅杰,等.参莲提取物对PM2.5染毒RAW264.7巨噬细胞损伤的保护作用[J].中国中药杂志, 2015, 40(10):1977-1983.DOI:10.4268/cjcmm20151025.
[30]曹柏营,姜秀娟,戚颖欣,等.藤本豆豆荚总黄酮对小鼠免疫功能的影响[J].食品与机械, 2017, 33(1):158-162. DOI:10.13652/j.issn.1003-5788.2017.01.036.
[2]WANGFulong,WANGZhenzhen,CHEN Yan. Advancesinthe researchofthemechanismandinterventionofaging[J].Chinese Journal of Cell Biology, 2012, 34(8):739-748.
[3]TRACHOOTHAMD,ZHANGH,ZHANGW,etal.Effective eliminationoffludarabine-resistantCLLcellsbyPEITCthrough aredox-mediatedmechanism[J].Blood,2008,112(5):1912-1922.DOI:10.1182/blood-2008-04-149815.
[4]WEST AP,BRODSKYIE,RAHNERC,etal.TLRsignalling augmentsmacrophagebactericidalactivitythroughmitochondrial ROS[J]. Nature, 2011, 472:476-480. DOI:10.1038/nature09973.
[5]卞梦瑶,方勇,裴斐,等.生姜油树脂对H2O2引起RAW264.7巨噬细胞损伤的保护作用[J].食品科学,2014,35(1):244-249.DOI:10.7506/spkx1002-6630-201323048.
[6]崔志文,黄琴,黄怡,等.鼠李糖乳酸杆菌对Caco-2细胞抗氧化功能的影响[J].中国农业科学, 2011, 44(23):4926-4932. DOI:10.3864/j.issn.0578-1752.2011.23.020.
[7]PLAZA L, CRESPO I, DE PASCUAL-TERESA S, et al. Impact of minimal processing on orange bioactive compounds during refrigerated storage[J].FoodChemistry,2011,124(2):646-651.DOI:10.1016/j.foodchem.2010.06.089.
[8]HUANG Renhua, XIA Renxue, HU Liming, et al. Antioxidant activity and oxygen-scavenging system in orange pulp during fruit ripening andmaturation[J].ScientiaHorticuvlturae,2007,113(2):166-172.DOI:10.1016/j.scienta.2007.03.010.
[9]JO S H, SON M K, KOH H J, et al. Control of mitochondrial redox balance and cellular defense against oxidative damage bymitochondrial NADP(+)-dependentisocitratedehydrogenase[J].TheJournalof Biological Chemistry, 2001, 276(19):16168-16176. DOI:10.1074/jbc.M010120200.
[10]LIAO H, BANBURY L K. Antioxidant activity of 45 Chinese herbs and the relationship with their TCM characteristics[J]. Evidencebased Complementary and Alternative Medicine, 2008, 5(4):429-434.DOI:10.1093/ecam/nem054.
[11]杨楠,贾晓斌,张振海,等.黄酮类化合物抗肿瘤活性及机制研究进展[J].中国中药杂志, 2015, 40(3):373-381. DOI:10.4268/cjcmm20140302.
[12]陈娟,张明华,章丽,等.牡丹皮苷/酚组分对糖尿病肾病大鼠肾损伤的保护作用及其机制研究[J].中国中药杂志, 2016, 41(11):1990-1998. DOI:10.4268/cjcmm20161104.
[13]赵伟,耿岩玲,崔莉,等.牡丹花黄酮类化学成分研究[J].中国现代中药,2016,18(3):303-306.DOI:10.13313/j.issn.1673-4890.2016.3.010.
[14]王顺利,任秀霞,薛璟祺,等.牡丹籽油成分、功效及加工工艺的研究进展[J].中国粮油学报, 2016, 31(3):139-146. DOI:10.3969/j.issn.1003-0174.2016.03.027.
[15]傅茂润,刘峰,赵海军,等.牡丹雄蕊的营养成分和抗氧化能力研究[J].中国食物与营养, 2011, 11(5):71-74. DOI:10.3969/j.issn.1006-9577.2011.05.017.
[16]李朝苹,陈立勇,阴英超,等.牡丹花蕊水提取液抗氧化功能研究[J].山东大学学报(医学版), 2015, 53(11):32-36. DOI:10.6040/j.issn.1671-7554.0.2015.252.
[17]代艳文,袁丁,万静枝,等.竹节参总皂苷通过NF-κB通路对LPS致RAW264.7巨噬细胞炎症的保护作用研究[J].中国中药杂志, 2014,39(11):2076-2080. DOI:10.4268/cjcmm20141126.
[18]邓洋,许秀举,于海平.啤酒对大鼠血清的LDH和T-AOC的影响[J].食品科技,2009,34(5):120-122.DOI:10.13684/j.cnki.spkj.2009.05.066.
[19]董亮,何永志,王远亮,等.超氧化物歧化酶(SOD)的应用研究进展[J].中国农业科技导报, 2013, 15(5):53-58. DOI:10.3969/j.issn.1008-0864.2013.05.08.
[20]WANG L Y, DING L, YU Z P, et al. Intracellular ROS scavenging and antioxidant enzyme regulating capacities of corn gluten meal-derived antioxidant peptides in HepG2 cells[J]. Food Research International,2016, 90(1):33-41. DOI:10.1016/j.foodres.2016.10.023.
[21]PODKOPAEVA D A, GRABOVICH M Y, DUBININA G A. Oxidative stress and antioxidant cell protection systems in the microaerophilic bacteriumSpirillumwinogradskii[J].Microbiologiya,2003,72(5):534-542. DOI:10.1023/A:1026082914661.
[22]BEDOYA-RAMíREZD,CILLA A,CONTRERAS-CALDERóNJ,etal.Evaluationoftheantioxidantcapacity,furancompoundsand cytoprotective-cytotoxiceffectsuponCaco-2cellsofcommercial Colombiancoffee[J].FoodChemistry,2017,219(1):364-372.DOI:10.1016/j.foodchem.2016.09.159.
[23]王文君,蔡教英,蒋艳,等.芦荟黄酮体内抗氧化活性及对CATmRNA表达的影响[J].中国食品学报,2013,13(10):13-18.DOI:10.16429/j.1009-7848.2013.10.003.
[24]苗明三,陈元朋,吴巍.姜黄素对大鼠血瘀性脑缺血模型血液流变学及脑匀浆LD、LDH和TchE水平的影响[J].中药药理与临床,2010, 26(1):29-31. DOI:10.13412/j.cnki.zyyl.2010.01.009.
[25]白健,师伟,秦鸿雁.巨噬细胞功能、活化和起源异质性在肝纤维化中作用的研究进展[J].细胞与分子免疫学杂志, 2017, 33(1):114-118. DOI:10.13423/j.cnki.cjcmi.008057.
[26]刘思思,李琦,孙立东,等.参莲提取物对LPS诱导的巨噬细胞炎症反应的影响[J].中国实验方剂学杂志, 2017, 23(10):85-91.DOI:10.13422/j.cnki.syfjx.2017100085.
[27]宮甜甜,黄少刚,张玥,等.巨噬细胞的极化及功能调控[J].解剖学报,2017,48(1):106-110.DOI:10.16098/j.issn.0529-1356.2017.01.019.
[28]袁芳.山楂叶总黄酮对H2O2诱导乳鼠心肌细胞氧化应激和细胞凋亡的影响[J].现代中西医结合杂志, 2016, 25(4):368-371; 405.DOI:10.3969/j.issn.1008-8849.2016.04.008.
[29]刘栩岑,李玉洁,王娅杰,等.参莲提取物对PM2.5染毒RAW264.7巨噬细胞损伤的保护作用[J].中国中药杂志, 2015, 40(10):1977-1983.DOI:10.4268/cjcmm20151025.
[30]曹柏营,姜秀娟,戚颖欣,等.藤本豆豆荚总黄酮对小鼠免疫功能的影响[J].食品与机械, 2017, 33(1):158-162. DOI:10.13652/j.issn.1003-5788.2017.01.036.