垂盆草总黄酮及异鼠李素对对乙酰氨基酚诱导的L02细胞损伤的保护作用
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摘要
目的:探讨垂盆草总黄酮及异鼠李素对对乙酰氨基酚(APAP)诱导的人正常肝细胞(L02)损伤的保护作用。方法:以不同浓度的APAP刺激L02细胞,制备损伤模型,筛选APAP的有效刺激浓度;复制APAP诱导的L02细胞损伤模型,用细胞增殖与活性试剂(CCK-8)检测细胞存活能力,分析盆草总黄酮及异鼠李素对APAP诱导的L02细胞活性的影响;检测细胞上清液中丙二醛(MDA),谷胱甘肽(GSH)含量,超氧化物歧化酶(SOD),谷胱甘肽过氧化物酶(GSH-Px)活性,丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)的含量。结果:与正常组比较,不同浓度的APAP处理的L02细胞活力显著降低,筛选出的造模条件为10 mmol·L-1APAP孵育24 h,模型组肝细胞上清液中MDA含量升高(P<0.05,P<0.01),GSH含量降低,SOD,GSH-Px活性降低,AST和ALT含量升高;与模型组比较,垂盆草总黄酮、异鼠李素均能显著提高APAP损伤细胞的存活率,降低MDA含量,提高GSH含量,提高SOD,GSH-Px活性,降低ALT和AST的释放量(P<0.05,P<0.01)。结论:垂盆草总黄酮及异鼠李素对APAP损伤的L02细胞具有较好的保护作用。
Objective: To study the protective effect of Sedi Herba total flavanones and isorhamnetin on the acetaminophen( APAP)-induced injury of human normal hepatocytes( L02). Method: L02 cells were stimulated with different concentrations of APAP to prepare the injury model, and the effective stimulating concentration of APAP was screened. APAP-induced L02 cell injury models were thencopied,and the effect of Sedi Herba total flavanones and isorhamnetin on the activity of L02 cells induced by APAP was analyzed by cell counting kit-8( CCK-8) method. The contents of malondialdehyde( MDA) and glutathione( GSH),activity of superoxide dismutase( SOD) and glutathione peroxidase( GSH-Px), and levels of alanine aminotransferase( ALT),andaspartate aminotransferase( AST) in supernatant were measured. Result: As compared with the normal group,the activity of L02 cells treated with different concentrations of APAP was significantly decreased,and the selected modeling condition was 10 mmol·L-1 for 24 h. The MDA content in the supernatant of the model group was increased and the GSH content was decreased,SOD and GSH-Px activity decreased,AST and ALT levels increased( P < 0. 05,P < 0. 01). As compared with the model group,the Sedi Herba total flavanones and isorhamnetin could significantly increase the survival rate of APAP-injured cells,decrease the content of MDA,increase the content of GSH,increase the activity of SOD and GSH-Px activity,and lower the release of ALT and AST( P <0. 05,P < 0. 01). Conclusion: The Sedi Herba total flavanones and isorhamnetin have a good protective effect on APAP-injured L02 cells.
Objective: To study the protective effect of Sedi Herba total flavanones and isorhamnetin on the acetaminophen( APAP)-induced injury of human normal hepatocytes( L02). Method: L02 cells were stimulated with different concentrations of APAP to prepare the injury model, and the effective stimulating concentration of APAP was screened. APAP-induced L02 cell injury models were thencopied,and the effect of Sedi Herba total flavanones and isorhamnetin on the activity of L02 cells induced by APAP was analyzed by cell counting kit-8( CCK-8) method. The contents of malondialdehyde( MDA) and glutathione( GSH),activity of superoxide dismutase( SOD) and glutathione peroxidase( GSH-Px), and levels of alanine aminotransferase( ALT),andaspartate aminotransferase( AST) in supernatant were measured. Result: As compared with the normal group,the activity of L02 cells treated with different concentrations of APAP was significantly decreased,and the selected modeling condition was 10 mmol·L-1 for 24 h. The MDA content in the supernatant of the model group was increased and the GSH content was decreased,SOD and GSH-Px activity decreased,AST and ALT levels increased( P < 0. 05,P < 0. 01). As compared with the model group,the Sedi Herba total flavanones and isorhamnetin could significantly increase the survival rate of APAP-injured cells,decrease the content of MDA,increase the content of GSH,increase the activity of SOD and GSH-Px activity,and lower the release of ALT and AST( P <0. 05,P < 0. 01). Conclusion: The Sedi Herba total flavanones and isorhamnetin have a good protective effect on APAP-injured L02 cells.
引文
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[14]王君明,崔瑛,王峥涛,等.超氧化物歧化酶参与肝损伤的研究进展[J].中国实验方剂学杂志,2011,17(7):265-269.
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[17]JIN S,DAI C,YANG Y,et al.Effect of hepatic blood inflow occlusion without hemihepatic artery control on apoptosis of liver cells in rats[J].Hepato Gastroenterol,2010,57(97):35-40.
[2]邵成雷.垂盆草化学成分及其药理临床研究[D].济南:山东大学,2006.
[3]潘金火,潘萍.垂盆草总黄酮的保肝降酶作用及其化学成分的鉴别研究[J].时珍国医国药,2010,21(8):1930-1934.
[4]董曦,孙桂波,罗云,等.异鼠李素对H2O2引起的H9C2细胞氧化应激损伤的保护作用研究[J].中国药理学通报,2015,31(6):853-860.
[5]梁容佳,陈俊秀,只德贤,等.异鼠李素对人肝微粒体CYPs活性及大鼠原代肝细胞毒性作用研究[J].药物评价研究,2017,40(5):627-632.
[6]AN H R,WU X Q.Antituberculosis drugs and hepatotoxicity[J].Chin J Antibiotics,2010,35(10):727-733.
[7]陈军宝,徐智,覃小艳,等.扑热息痛肝损伤机制的研究进展[J].实用医学杂志,2012,28(20):3479-3480.
[8]赵慧,吕佳,曹燕妮,等.甘草酸铵对对乙酰氨基酚致人肝细胞损伤的保护作用研究[J].中南药学,2017,15(3):301-304.
[9]贺兰芝,孟雅坤,韩延忠,等.木犀草素对对乙酰氨基酚诱导的L02肝细胞损伤的保护作用[J].中国中药杂志,2016,41(22):4234-4239.
[10]魏鹏,刘伟霞,贾凤兰,等.8-甲氧补骨脂素对对乙酰氨基酚致小鼠急性肝损伤的保护作用[J].中国现代应用药学,2012,29(8):682-686.
[11]Jaeschke H,Mc Gill M R,Ramachandran A.Oxidant stress,mitochondria,and cell death mechanisms in drug-induced liver injury:lessons learned from acetaminophen hepatotoxicity[J].Drug Metab Rev,2012,44(1):88-106.
[12]刘霞飞,吴骁,杨方秀,等.正柴胡饮对对乙酰氨基酚所致小鼠急性肝损伤的保护作用[J].中国药理学与毒理学杂志,2017,31(1):101-111.
[13]鲁合军.垂盆草水煎液对4种肝损伤模型小鼠的保肝作用[J].中医学报,2017,32(3):409-414.
[14]王君明,崔瑛,王峥涛,等.超氧化物歧化酶参与肝损伤的研究进展[J].中国实验方剂学杂志,2011,17(7):265-269.
[15]王英,贾杰.AST/ALT比值在各种肝病中的变化和意义[J].中国热带医学,2001,1(3):222-223.
[16]Hsiao G,LIN Y H,LIN C H,et al.The protective effects of PMC against chronic carbon tetrachlorideinduced hepatotoxicity in vivo[J].Biol Pharm Bull,2001,24(11):1271-1276.
[17]JIN S,DAI C,YANG Y,et al.Effect of hepatic blood inflow occlusion without hemihepatic artery control on apoptosis of liver cells in rats[J].Hepato Gastroenterol,2010,57(97):35-40.