表没食子儿茶素没食子酸酯对黄曲霉毒素B_1致肝细胞氧化损伤的保护作用
|
摘要
目的:观察表没食子儿茶素没食子酸酯(EGCG)对黄曲霉毒素B_1(AFB_1)致人正常肝细胞(HL7702细胞)氧化损伤的影响。方法:将HL7702细胞分为正常对照组、溶剂对照组(0.3%DMSO)、AFB_1染毒组和低、中、高剂量EGCG组(5μg/mL、10μg/mL、20μg/mL)。AFB_1染毒组用60μg/mL AFB_1作用细胞24 h,建立肝HL7702细胞氧化应激模型。低、中、高剂量EGCG组预先用相应浓度EGCG处理细胞1 h,再加入60μg/mL AFB_1作用24 h。用CCK-8法检测细胞存活率,DCFH-DA荧光探针法检测细胞内活性氧(ROS),并检测细胞培养上清液中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、乳酸脱氢酶(LDH)以及受损细胞内超氧化物歧化酶(SOD)、丙二醛(MDA)的含量。结果:与对照组相比,AFB_1染毒组ALT、AST、MDA及ROS水平明显升高,肝细胞存活率、SOD酶活性明显降低,差异均有统计学意义(均P<0.05);与AFB_1染毒组相比,EGCG处理可使ALT、AST、MDA及ROS水平明显降低,肝细胞存活率、SOD活性明显升高(均P<0.05)。结论:EGCG可明显减轻AFB_1诱导的肝细胞损伤,其作用机制可能与抗氧化损伤有关。
Objective:To study the effect of epigallocatechin gallate(EGCG) on human hepatic cell line(HL7702) damaged by aflatoxin B_1(AFB_1).Methods:HL7702 cells were divided into control group,solvent control group(0.3% DMSO),AFB_1 group,and low-,medium-,and high-dose(5,10 and 20 μg/mL) of EGCG groups.The cells in AFB_1 group and EGCG groups were treated with 60 μg/mL AFB_1 for 24 h.EGCG groups cells were treated with different concentrations of EGCG at 1 h before the addition of AFB_1.CCK-8 assay was used to detect cell viability.The reactive oxygen species(ROS) in cells was measured by DCFH-DA fluorescence probe.The contents of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and lactate dehydrogenase(LDH) in cells supernatant as well as the intracellular superoxide dismutase(SOD) and malondialdehyde(MDA) in cells were determined.Results:Compared with the control group,the levels of ALT,AST,MDA and ROS in AFB_1 group were significantly increased,while the hepatocytes survival rate and the activity of SOD were decreased(P<0.05).All these effects of AFB_1 were reversed by pretreatment with EGCG(P<0.05).Conclusion:EGCG could reduce hepatocytes injury induced by AFB_1,and the mechanism might be related to antioxidant damage.
Objective:To study the effect of epigallocatechin gallate(EGCG) on human hepatic cell line(HL7702) damaged by aflatoxin B_1(AFB_1).Methods:HL7702 cells were divided into control group,solvent control group(0.3% DMSO),AFB_1 group,and low-,medium-,and high-dose(5,10 and 20 μg/mL) of EGCG groups.The cells in AFB_1 group and EGCG groups were treated with 60 μg/mL AFB_1 for 24 h.EGCG groups cells were treated with different concentrations of EGCG at 1 h before the addition of AFB_1.CCK-8 assay was used to detect cell viability.The reactive oxygen species(ROS) in cells was measured by DCFH-DA fluorescence probe.The contents of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and lactate dehydrogenase(LDH) in cells supernatant as well as the intracellular superoxide dismutase(SOD) and malondialdehyde(MDA) in cells were determined.Results:Compared with the control group,the levels of ALT,AST,MDA and ROS in AFB_1 group were significantly increased,while the hepatocytes survival rate and the activity of SOD were decreased(P<0.05).All these effects of AFB_1 were reversed by pretreatment with EGCG(P<0.05).Conclusion:EGCG could reduce hepatocytes injury induced by AFB_1,and the mechanism might be related to antioxidant damage.
引文
[1] KASOJU A,NARASU M L,MUVVA C,et al.Prediction of the three-dimensional structure of aflatoxin of Aspergillus flavus by homology modelling[J].Bioinformation,2012,8(14):684-686.
[2] YANAN A,XIAOCHEN S,XUDONG T,et al.Aflatoxin B1induces reactive oxygen species-mediated autophagy and extracellular trap formation in macrophages[J].Front Cell Infect Microbiol,2017(7):53.
[3] PETRUZZIELLO A.Epidemiology ofhepatitis B virus (HBV) and hepatitis C virus (HCV) related hepatocellular carcinoma[J].Open Virol J,2018,12(Suppl-1,M3):26-32.
[4] YILMAZ S,KAYA E,COMAKLI S.Vitamin E (α tocopherol) attenuates toxicity and oxidative stress induced by aflatoxin in rats[J].Adv Clin Exp Med,2017,26(6):907-917.
[5] PAN H,CHEN J,SHEN K,et al.Mitochondrialmodulation by epigallocatechin 3-gallate ameliorates cisplatin induced renal injury through decreasing oxidative/nitrative stress,inflammation and NF-kB in mice[J].PLoS One,2015,10(4):e0124775.
[6] ALEKSANDRA N,MOHD R,TATIANA K,et al.Anticancer efficacy of polyphenols and their combinations[J].Nutrients,2016,8(9):E552.
[7] 赵静芳,邓志杰,肖德强,等.表没食子儿茶素没食子酸酯对黄曲霉毒素B1诱导大鼠急性肝损伤的抗氧化作用[J].实用医学杂志,2018,34(11):1753-1756,1761.
[8] ROTIMI O A,ROTIMI S O,OLUWAFEMI F,et al.Oxidative stress in extrahepatic tissues of rats co-exposed to aflatoxin B1 and low protein diet[J].Toxicol Res,2018,34(3):211-220.
[9] DOHNAL V,WU Q,KAMIL KUCA.Metabolism of aflatoxins:Key enzymes and interindividual as well as interspecies differences[J].Arch Toxicol,2014,88(9):1635-1644.
[10] MARIN D E,TARANU I.Overview on aflatoxins and oxidative stress[J].Toxin Rev,2012,31(3-4):32-43.
[11] ROTIMI O A,ROTIMI S O,OLUWAFEMI F,et al.Coexistence ofaflatoxicosis with protein malnutrition worsens hepatic oxidative damage in rats[J].J Biochem Mol Toxicol,2016,30(6):269-276.
[12] ABDEL-AZIEM SH,HASSAN A M,EL-DENSHARY E S,et al.Ameliorative effects of thyme and calendula extracts alone or in combination against aflatoxins-induced oxidative stress and genotoxicity in rat liver[J].Cytotechnology,2014,66(3):457-470.
[13] KUMAR M B,KUMAR T S.Fisetinmodulates antioxidant enzymes and inflammatory factors to inhibit aflatoxin-B1 induced hepatocellular carcinoma in rats[J].Oxid Med Cell Longev,2016(2016):1972793.
[14] 刘婷婷,孟馨.表没食子儿茶素没食子酸酯抗氧化作用机制的研究进展[J].现代药物与临床,2016,31(6):919-923.
[15] VALENCIA A M,ABRANTES M A,HASAN J,et al.Reactiveoxygen species,biomarkers of microvascular maturation and alveolarization,and antioxidants in oxidative lung injury[J].React Oxyg Species (Apex),2018,6(18):373-388.
[16] YUNJIE X,WEINAN G,YONG Z,et al.ABT737 reverses cisplatin resistance by targeting glucose metabolism of human ovarian cancer cells[J].Int J Oncol,2018,53:1055-1068.
[17] QI S,FENG Z,LI Q,et al.Inhibition of ROS-mediated activation Src-MAPK/AKT signaling by orientin alleviates H2O2-induced apoptosis in PC12 cells[J].Drug Des Devel Ther,2018(12):3973-3984.
[18] 朱俊杰,徐静.大青叶提取物对HepG2细胞酒精性损伤的保护作用[J].食品研究与开发,2018,39(15):161-167.
[19] 张笑天,郑晓瑛.氧化自由基清除剂超氧化物歧化酶与疾病[J].中国公共卫生,2014,30(10):1349-1352.
[2] YANAN A,XIAOCHEN S,XUDONG T,et al.Aflatoxin B1induces reactive oxygen species-mediated autophagy and extracellular trap formation in macrophages[J].Front Cell Infect Microbiol,2017(7):53.
[3] PETRUZZIELLO A.Epidemiology ofhepatitis B virus (HBV) and hepatitis C virus (HCV) related hepatocellular carcinoma[J].Open Virol J,2018,12(Suppl-1,M3):26-32.
[4] YILMAZ S,KAYA E,COMAKLI S.Vitamin E (α tocopherol) attenuates toxicity and oxidative stress induced by aflatoxin in rats[J].Adv Clin Exp Med,2017,26(6):907-917.
[5] PAN H,CHEN J,SHEN K,et al.Mitochondrialmodulation by epigallocatechin 3-gallate ameliorates cisplatin induced renal injury through decreasing oxidative/nitrative stress,inflammation and NF-kB in mice[J].PLoS One,2015,10(4):e0124775.
[6] ALEKSANDRA N,MOHD R,TATIANA K,et al.Anticancer efficacy of polyphenols and their combinations[J].Nutrients,2016,8(9):E552.
[7] 赵静芳,邓志杰,肖德强,等.表没食子儿茶素没食子酸酯对黄曲霉毒素B1诱导大鼠急性肝损伤的抗氧化作用[J].实用医学杂志,2018,34(11):1753-1756,1761.
[8] ROTIMI O A,ROTIMI S O,OLUWAFEMI F,et al.Oxidative stress in extrahepatic tissues of rats co-exposed to aflatoxin B1 and low protein diet[J].Toxicol Res,2018,34(3):211-220.
[9] DOHNAL V,WU Q,KAMIL KUCA.Metabolism of aflatoxins:Key enzymes and interindividual as well as interspecies differences[J].Arch Toxicol,2014,88(9):1635-1644.
[10] MARIN D E,TARANU I.Overview on aflatoxins and oxidative stress[J].Toxin Rev,2012,31(3-4):32-43.
[11] ROTIMI O A,ROTIMI S O,OLUWAFEMI F,et al.Coexistence ofaflatoxicosis with protein malnutrition worsens hepatic oxidative damage in rats[J].J Biochem Mol Toxicol,2016,30(6):269-276.
[12] ABDEL-AZIEM SH,HASSAN A M,EL-DENSHARY E S,et al.Ameliorative effects of thyme and calendula extracts alone or in combination against aflatoxins-induced oxidative stress and genotoxicity in rat liver[J].Cytotechnology,2014,66(3):457-470.
[13] KUMAR M B,KUMAR T S.Fisetinmodulates antioxidant enzymes and inflammatory factors to inhibit aflatoxin-B1 induced hepatocellular carcinoma in rats[J].Oxid Med Cell Longev,2016(2016):1972793.
[14] 刘婷婷,孟馨.表没食子儿茶素没食子酸酯抗氧化作用机制的研究进展[J].现代药物与临床,2016,31(6):919-923.
[15] VALENCIA A M,ABRANTES M A,HASAN J,et al.Reactiveoxygen species,biomarkers of microvascular maturation and alveolarization,and antioxidants in oxidative lung injury[J].React Oxyg Species (Apex),2018,6(18):373-388.
[16] YUNJIE X,WEINAN G,YONG Z,et al.ABT737 reverses cisplatin resistance by targeting glucose metabolism of human ovarian cancer cells[J].Int J Oncol,2018,53:1055-1068.
[17] QI S,FENG Z,LI Q,et al.Inhibition of ROS-mediated activation Src-MAPK/AKT signaling by orientin alleviates H2O2-induced apoptosis in PC12 cells[J].Drug Des Devel Ther,2018(12):3973-3984.
[18] 朱俊杰,徐静.大青叶提取物对HepG2细胞酒精性损伤的保护作用[J].食品研究与开发,2018,39(15):161-167.
[19] 张笑天,郑晓瑛.氧化自由基清除剂超氧化物歧化酶与疾病[J].中国公共卫生,2014,30(10):1349-1352.