蟛蜞菊内酯对对乙酰氨基酚致小鼠急性肝损伤的保护作用
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摘要
目的探究蟛蜞菊内酯对对乙酰氨基酚引起的小鼠急性肝损伤的保护作用。方法采用小鼠腹腔注射对乙酰氨基酚模拟肝损伤模型,观察蟛蜞菊内酯对肝的保护作用。将BALB/c小鼠随机分为对照组、模型组、蟛蜞菊内酯低剂量组和高剂量组(n=8)。通过检测小鼠血清中的丙氨酸转移酶(ALT)、天冬氨酸转移酶(AST);肝组织匀浆中的丙二醛(MDA)、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GSH-PX)和超氧化物歧化酶(SOD)指标,血清ELIA检测TNF-α、IL-6,结合肝的组织病理学检查,观察蟛蜞菊内酯对对乙酰氨基酚所致肝损伤的保护作用。结果模型组小鼠血清中的AST、ALT、GSH、MDA、IL-6以及TNF-α水平水平明显高于对照组(P<0. 01),有统计学意义;蟛蜞菊内酯低剂量组和高剂量组的AST、ALT以及肝组织匀浆液中SOD和GSH-PX水平明显要高于模型组,且肝组织匀浆液中MDA、GSH水平以及小鼠血清中TNF-α、IL-6水平明显低于模型组(P<0. 01),有统计学意义;模型组的GSH-PX和SOD水平明显低于对照组(P<0. 01),有统计学意义。组织病理学苏木素-伊红(hematoxylin and eosin,HE)和细胞凋亡染色(Td T-mediated DUTP nick end labeling,TUNEL)显示蟛蜞菊内酯可明显减轻肝组织的坏死和凋亡。结论蟛蜞菊内酯对对乙酰氨基酚引起的急性肝损伤有明显的保护作用。
Objective To investigate the protective effect of wedelolactone( Wed) against acetaminophen( APAP)-induced acute liver in mice. Methods A mouse model of APAP-induced liver injury was established. Thirty-two male BALB/c mice were randomly divided into normal,APAP,10 mg/kg Wed and 20 mg/kg Wed groups( n = 8). A colorimetric method was used to assay the levels of serum alanine aminotransferase( ALT),aspartate aminotransferase( AST),malondialdehyde( MDA),glutathione( GSH),glutathione peroxidase( GSH-PX) and superoxide dismutase( SOD) in mouse liver homogenates,and the serum levels of tumor necrosis factor( TNF-α) and interleukin-6( IL-6).Results In the model group,the serum levels of AST,ALT,IL-6 and TNF-α,and GSH and MDA in the liver homogenates were significantly higher than those in the control group( P < 0. 01). The levels of AST,ALT,SOD and GSH-PX in liver tissue homogenates of the low-and high-dose Wed groups were significantly higher than those in the model group( P <0. 01),and the levels of MDA and GSH in liver tissue homogenates and TNF-α and IL-6 in the serum of mice were significantly higher than those in the model group( P < 0. 01). The levels of GSH-PX and SOD in the model group were significantly lower than those in the control group( P < 0. 01). Histopathological examination using HE and TUNEL staining showed that Wed significantly reduced liver necrosis and apoptosis. Conclusions Wed has a protective effect on APAP-induced acute liver injury.
Objective To investigate the protective effect of wedelolactone( Wed) against acetaminophen( APAP)-induced acute liver in mice. Methods A mouse model of APAP-induced liver injury was established. Thirty-two male BALB/c mice were randomly divided into normal,APAP,10 mg/kg Wed and 20 mg/kg Wed groups( n = 8). A colorimetric method was used to assay the levels of serum alanine aminotransferase( ALT),aspartate aminotransferase( AST),malondialdehyde( MDA),glutathione( GSH),glutathione peroxidase( GSH-PX) and superoxide dismutase( SOD) in mouse liver homogenates,and the serum levels of tumor necrosis factor( TNF-α) and interleukin-6( IL-6).Results In the model group,the serum levels of AST,ALT,IL-6 and TNF-α,and GSH and MDA in the liver homogenates were significantly higher than those in the control group( P < 0. 01). The levels of AST,ALT,SOD and GSH-PX in liver tissue homogenates of the low-and high-dose Wed groups were significantly higher than those in the model group( P <0. 01),and the levels of MDA and GSH in liver tissue homogenates and TNF-α and IL-6 in the serum of mice were significantly higher than those in the model group( P < 0. 01). The levels of GSH-PX and SOD in the model group were significantly lower than those in the control group( P < 0. 01). Histopathological examination using HE and TUNEL staining showed that Wed significantly reduced liver necrosis and apoptosis. Conclusions Wed has a protective effect on APAP-induced acute liver injury.
引文
[1]张倩,刘江宁,秦川.乙型病毒性肝炎动物模型的比较分析[J].中国比较医学杂志,2017,27(06):72-76.
[2]汪倩,徐瑞娟,杨劲.对乙酰氨基酚肝毒性机理及药物干预靶点[J].药学与临床研究,2011,19(3):247
[3]李三强,韩红梅,卢华杰,等.解整合素-金属蛋白酶9在对乙酰氨基酚诱导的小鼠急性肝损伤中的表达分析[J].中国临床药理学杂志,2015,31(12):1143-1145.
[4]李大伟,陆天飞,华相伟,等.对乙酰氨基酚诱导的小鼠药物性肝损伤的模型研究[J].中国细胞生物学学报,2014,36(6):805-809.
[5]潘家琪,宋丹军,李鹏旭,等.对乙酰氨基酚肝毒性机制与防治研究新进展[J].中国药理学与毒理学杂志,2014,28(4):618-624.
[6] Govindachari TR,Nagarajan K,Pai BR. Chemical examination of Wedelia calendulacea. Part 1. Structure of wedelolactone[J].J Chem Soc,1956:629-632.
[7] Mors WB,Nascimento MC,Parente JP,et al. Neutralization of lethal and myotoxic activities of South American rattlesnake venom by extracts and constituents of the plant Eclipta prostrate(Asteraceae)[J]. Toxicon,1989,27(9):1003-1009.
[8] Wagner H,Geyer B,Kisp Y,et al. Coumestans as the main active principles of the liver drugs Eclipta alba and wedelia calenulaceae[J]. Planta Med,1986,10(5):370-374.
[9] Kobori M,Yang Z,Gong D,et al. Wedelolactone suppresses LPS-induced caspase-11 expression by directly inhibiting the IKK complex[J]. Cell Death Differ,2004,11(1):123-130.
[10]陈杰,许静,罗子玲,等.蟛蜞菊内酯对脂多糖诱导RAW264. 7细胞IL-6、TNF-α及NF-κB的影响[J].时珍国医国药,2012,23(9):2166.
[11] Xia Y,Chen J,Cao Y,et al. Wedelolactone exhibits antifibrotic effects on human hepatic stellate cell line LX-2[J].Eur J Pharmacol,2013,714(1-3):105-111.
[12] Bleibel W,Kim S,D’Silva K,et al. Drug-induced liver injury:review article[J]. Dig Dis Sci,2007,52(10):2463-2471.
[13]刘冰,姜树民.中药制剂治疗急性肝损伤临床观察[J].辽宁中医药大学学报,2014,16(11):101-103.
[14]周琼,刘芳萍,赵玉林,等.四氯化碳致小鼠急性肝损伤动物模型建立方法的[J].东北农业大学学报,2012,43(6):77-81.
[15]邢旺兴,陈斌,宓鹤鸣,等.乌骨藤辨考[J].中药材,2003,26(7):524-526.
[16]邢旺兴,唐婷,谷娜,等.抗肿瘤中药通光藤的药理作用及其作用机理初步研究[J].健康研究,2011,31(1):12-16
[17] Ghosh J,Das J,Manna P,et al. Acetaminophen induced renal injury via oxidative stress and TNF-alpha production:therapeutic potential of arjunolic acid[J]. Toxicology,2010,268(1-2):8-18
[18] Ghosh J, Das J, Manna P, et al. Arjunolic acid, a triterpenoidsaponin,prevents acetaminophen(APAP)-induced liver and hepatocyte injury via the inhibition of APAP bioactivationand JNK-mediated mitochondrial protection[J].Free Radic Biol Med,2010,48(4):535-553.
[19] Terneus MV,Kiningham KK,Carpenter AB,et al. Comparison of S-Adenosyl-L-methionine and N-acetylcysteine protective effects on acetaminophen hepatic toxicity[J]. J Pharmacol Exp Ther,2007,320(1):99-107.
[20] Latchoumycandane C,Goh CW,Ong MM,et al. Mitochondrial protection by the JNK inhibitor leflunomide rescues micefrom acetaminophen-induced liver injury[J]. Hepatology,2007,45(2):412-421.
[21] Mullins ME,Schmidt RU,Jand TB,et al. What is the rate of adverse events with intravenous versus oral N-acetylcysteine in pediatric patients[J]. Ann Emerg Med,2004,44(5):547-548.
[22] Schmidt LE,Dalhoff K. Risk factors in the development of adverse reactions to N-acetylcysteine in patients with paracetamol poisoning[J].Br J Clin Pharmacol,2001,51(1):87-91.
[23] Adewusi EA,Afolayan AJ. A review of natural products with hepatoprotective activity[J]. J Med Plants Res,2010,4(13):1318-1334.
[24]何伟,宋莎莎,袁平凡,等.雷公藤红素对二乙基亚硝胺诱导的大鼠肝纤维化的治疗作用及机制[J].中国药理学通报,2013,29(4):519-524
[25]陈彤,李丽洪,陈小河,梁金平,杨立明.紫甘薯叶超声波提取工艺优化及其对小鼠肝脏保护作用研究[J].天然产物研究与开发,2016,28(08):1212-1218.
[26] Hsu YW,Tsai CF, Chen WK,et al. Protective effects of seabuckthorn(Hippophae rhamnoides L)seed oil against carbon tetrachloride-induced hepatotoxicity in mice[J]. Food Chem Toxicol,2009,47(9):2281-2288.
[27]李雅莉,赵玲,张丽,张兰,李林.复原再造胶囊对全脑缺血小鼠学习记忆及脑内氧化应激的影响[J].中国比较医学杂志,2015,25(08):1-5+83.
[28] Jaeschke H. Role of inflammation in the mechanism of acetaminophen-induced hepatotocicity[J]. Expert Opin Drug Mrtab Toxicol,2005,1(3):389-397.
[29] Ishida Y,Kondo T,Ohshima T,et al. A pivotal involvement of IFN-gamma in the pathogenesis of acetaminophen-induced acute liver injury[J].. FASEB J,2002,16(10):1227-1236.
[30] Masubuchi Y,Bourdi M,Reilly TP,et al. Role of interleukin-6in hepatic heat shock protein expression and protection against acetaminophen-induced liver disease[J]. Biochem Biophys Res Commun,2003,304(1):207-212.
[2]汪倩,徐瑞娟,杨劲.对乙酰氨基酚肝毒性机理及药物干预靶点[J].药学与临床研究,2011,19(3):247
[3]李三强,韩红梅,卢华杰,等.解整合素-金属蛋白酶9在对乙酰氨基酚诱导的小鼠急性肝损伤中的表达分析[J].中国临床药理学杂志,2015,31(12):1143-1145.
[4]李大伟,陆天飞,华相伟,等.对乙酰氨基酚诱导的小鼠药物性肝损伤的模型研究[J].中国细胞生物学学报,2014,36(6):805-809.
[5]潘家琪,宋丹军,李鹏旭,等.对乙酰氨基酚肝毒性机制与防治研究新进展[J].中国药理学与毒理学杂志,2014,28(4):618-624.
[6] Govindachari TR,Nagarajan K,Pai BR. Chemical examination of Wedelia calendulacea. Part 1. Structure of wedelolactone[J].J Chem Soc,1956:629-632.
[7] Mors WB,Nascimento MC,Parente JP,et al. Neutralization of lethal and myotoxic activities of South American rattlesnake venom by extracts and constituents of the plant Eclipta prostrate(Asteraceae)[J]. Toxicon,1989,27(9):1003-1009.
[8] Wagner H,Geyer B,Kisp Y,et al. Coumestans as the main active principles of the liver drugs Eclipta alba and wedelia calenulaceae[J]. Planta Med,1986,10(5):370-374.
[9] Kobori M,Yang Z,Gong D,et al. Wedelolactone suppresses LPS-induced caspase-11 expression by directly inhibiting the IKK complex[J]. Cell Death Differ,2004,11(1):123-130.
[10]陈杰,许静,罗子玲,等.蟛蜞菊内酯对脂多糖诱导RAW264. 7细胞IL-6、TNF-α及NF-κB的影响[J].时珍国医国药,2012,23(9):2166.
[11] Xia Y,Chen J,Cao Y,et al. Wedelolactone exhibits antifibrotic effects on human hepatic stellate cell line LX-2[J].Eur J Pharmacol,2013,714(1-3):105-111.
[12] Bleibel W,Kim S,D’Silva K,et al. Drug-induced liver injury:review article[J]. Dig Dis Sci,2007,52(10):2463-2471.
[13]刘冰,姜树民.中药制剂治疗急性肝损伤临床观察[J].辽宁中医药大学学报,2014,16(11):101-103.
[14]周琼,刘芳萍,赵玉林,等.四氯化碳致小鼠急性肝损伤动物模型建立方法的[J].东北农业大学学报,2012,43(6):77-81.
[15]邢旺兴,陈斌,宓鹤鸣,等.乌骨藤辨考[J].中药材,2003,26(7):524-526.
[16]邢旺兴,唐婷,谷娜,等.抗肿瘤中药通光藤的药理作用及其作用机理初步研究[J].健康研究,2011,31(1):12-16
[17] Ghosh J,Das J,Manna P,et al. Acetaminophen induced renal injury via oxidative stress and TNF-alpha production:therapeutic potential of arjunolic acid[J]. Toxicology,2010,268(1-2):8-18
[18] Ghosh J, Das J, Manna P, et al. Arjunolic acid, a triterpenoidsaponin,prevents acetaminophen(APAP)-induced liver and hepatocyte injury via the inhibition of APAP bioactivationand JNK-mediated mitochondrial protection[J].Free Radic Biol Med,2010,48(4):535-553.
[19] Terneus MV,Kiningham KK,Carpenter AB,et al. Comparison of S-Adenosyl-L-methionine and N-acetylcysteine protective effects on acetaminophen hepatic toxicity[J]. J Pharmacol Exp Ther,2007,320(1):99-107.
[20] Latchoumycandane C,Goh CW,Ong MM,et al. Mitochondrial protection by the JNK inhibitor leflunomide rescues micefrom acetaminophen-induced liver injury[J]. Hepatology,2007,45(2):412-421.
[21] Mullins ME,Schmidt RU,Jand TB,et al. What is the rate of adverse events with intravenous versus oral N-acetylcysteine in pediatric patients[J]. Ann Emerg Med,2004,44(5):547-548.
[22] Schmidt LE,Dalhoff K. Risk factors in the development of adverse reactions to N-acetylcysteine in patients with paracetamol poisoning[J].Br J Clin Pharmacol,2001,51(1):87-91.
[23] Adewusi EA,Afolayan AJ. A review of natural products with hepatoprotective activity[J]. J Med Plants Res,2010,4(13):1318-1334.
[24]何伟,宋莎莎,袁平凡,等.雷公藤红素对二乙基亚硝胺诱导的大鼠肝纤维化的治疗作用及机制[J].中国药理学通报,2013,29(4):519-524
[25]陈彤,李丽洪,陈小河,梁金平,杨立明.紫甘薯叶超声波提取工艺优化及其对小鼠肝脏保护作用研究[J].天然产物研究与开发,2016,28(08):1212-1218.
[26] Hsu YW,Tsai CF, Chen WK,et al. Protective effects of seabuckthorn(Hippophae rhamnoides L)seed oil against carbon tetrachloride-induced hepatotoxicity in mice[J]. Food Chem Toxicol,2009,47(9):2281-2288.
[27]李雅莉,赵玲,张丽,张兰,李林.复原再造胶囊对全脑缺血小鼠学习记忆及脑内氧化应激的影响[J].中国比较医学杂志,2015,25(08):1-5+83.
[28] Jaeschke H. Role of inflammation in the mechanism of acetaminophen-induced hepatotocicity[J]. Expert Opin Drug Mrtab Toxicol,2005,1(3):389-397.
[29] Ishida Y,Kondo T,Ohshima T,et al. A pivotal involvement of IFN-gamma in the pathogenesis of acetaminophen-induced acute liver injury[J].. FASEB J,2002,16(10):1227-1236.
[30] Masubuchi Y,Bourdi M,Reilly TP,et al. Role of interleukin-6in hepatic heat shock protein expression and protection against acetaminophen-induced liver disease[J]. Biochem Biophys Res Commun,2003,304(1):207-212.