老鼠簕生物碱A对小鼠药物性肝损伤的保护作用
|
摘要
目的:观察老鼠簕生物碱A(AAIA)对对乙酰氨基酚引起肝损伤模型的影响。方法:将小鼠随机分成正常组、模型组、联苯双酯组(联苯双酯,150 mg·kg~(-1))及AAIA高、中、低剂量(200,100,50 mg·kg~(-1))组,每组10只,连续灌胃给药10 d,末次给药后禁食不禁水8 h,除正常组外,其余各组给予腹腔注射275 mg·kg~(-1)对乙酰氨基酚,诱导小鼠急性肝损伤模型。6 h后,进行眼球取血,称量小鼠体质量、肝脏、脾脏、肾脏、胸腺质量,计算相应脏器指数,使用试剂盒检测血清中丙氨酸氨基转移酶(ALT),天冬氨酸氨基转移酶(AST)的含量,紫外分光光度法测定肝匀浆中一氧化氮(NO),诱导型一氧化氮合酶(i NOS)的含量,蛋白免疫印迹法(Western blot)测定细胞外调节蛋白激酶(ERK1/2),磷酸化细胞外调节蛋白激酶(p-ERK1/2)的表达。结果:与正常组比较,模型组的肝脏指数,血清中AST,ALT,肝组织匀浆NO,iNOS水平及p-ERK1/2在肝脏中的蛋白表达显著增高(P <0. 01);与模型组比较,各给药组均能降低对乙酰氨基酚急性肝损伤小鼠肝脏指数,血清AST,ALT水平,肝组织匀浆中NO,iNOS的产生,肝脏p-ERK1/2蛋白表达(P <0. 01);所有注射对乙酰氨基酚组的脾脏、肾脏及胸腺无明显变化,差异无统计学意义。结论:AAIA可能通过降低AST,ALT的水平,下调NO,iNOS的表达,减少p-ERK1/2的表达,对小鼠药物性肝损伤起到保护作用。
Objective: To observe the effect of alkaloid A from Acanthi Ilicifolii Herba seu Radix(AAIA) on liver injury model caused by acetaminophen. Method: Mice were randomly divided into normal group,model group,positive drug group(bifendate,150 mg·kg~(-1)) and high,medium and low-dose AAIA groups(200,100,50 mg·kg~(-1)),with 10 in each group. They were given drugs by gavage for 10 days,and fasted for 8 hours after the last administration. Except the normal group,the other groups were intraperitoneally injected with275 mg·kg~(-1) acetaminophen to induce acute liver injury model in mice. Six hours later,blood was taken from the eyeball. The body,liver,spleen,kidney and thymus were weighed,and then the corresponding organ indexes were calculated. The kits were used to detect the contents of aspartate aminotransferase(AST) and alanine aminotransferase(ALT) in serum. The contents of nitric oxide(NO) and inducible nitric oxide synthase(i NOS)in liver homogenate were determined by ultraviolet spectrophotometry,and the expressions of extracellular regulated protein kinases(ERK1/2) and phosphorylated extracellular regulatory protein kinase(p-ERK1/2) were determined by Western blot. Result: Compared with the normal group,the liver index,serum AST and ALT levels,the production of NO and i NOS in liver homogenate,the expression of p-ERK1/2 protein in liver of the model group increased significantly(P < 0. 01). Compared with the model group,all of drug groups could reduce the liver index of mice with acute liver injury induced by acetaminophen,the levels of serum AST and ALT,the production of NO and i NOS in liver and homogenate and the expression of p-ERK1/2 protein in liver(P < 0. 01).however,it had no significant effect on spleen,kidney and thymus of all acetaminophen injection groups.Conclusion: AAIA may protect mice from drug-induced liver injury by reducing AST and ALT levels,downregulating the expressions of NO and i NOS,and reducing the expression of protein p-ERK1/2.
Objective: To observe the effect of alkaloid A from Acanthi Ilicifolii Herba seu Radix(AAIA) on liver injury model caused by acetaminophen. Method: Mice were randomly divided into normal group,model group,positive drug group(bifendate,150 mg·kg~(-1)) and high,medium and low-dose AAIA groups(200,100,50 mg·kg~(-1)),with 10 in each group. They were given drugs by gavage for 10 days,and fasted for 8 hours after the last administration. Except the normal group,the other groups were intraperitoneally injected with275 mg·kg~(-1) acetaminophen to induce acute liver injury model in mice. Six hours later,blood was taken from the eyeball. The body,liver,spleen,kidney and thymus were weighed,and then the corresponding organ indexes were calculated. The kits were used to detect the contents of aspartate aminotransferase(AST) and alanine aminotransferase(ALT) in serum. The contents of nitric oxide(NO) and inducible nitric oxide synthase(i NOS)in liver homogenate were determined by ultraviolet spectrophotometry,and the expressions of extracellular regulated protein kinases(ERK1/2) and phosphorylated extracellular regulatory protein kinase(p-ERK1/2) were determined by Western blot. Result: Compared with the normal group,the liver index,serum AST and ALT levels,the production of NO and i NOS in liver homogenate,the expression of p-ERK1/2 protein in liver of the model group increased significantly(P < 0. 01). Compared with the model group,all of drug groups could reduce the liver index of mice with acute liver injury induced by acetaminophen,the levels of serum AST and ALT,the production of NO and i NOS in liver and homogenate and the expression of p-ERK1/2 protein in liver(P < 0. 01).however,it had no significant effect on spleen,kidney and thymus of all acetaminophen injection groups.Conclusion: AAIA may protect mice from drug-induced liver injury by reducing AST and ALT levels,downregulating the expressions of NO and i NOS,and reducing the expression of protein p-ERK1/2.
引文
[1]杨盛昌,陆文勋,邹祯,等.中国红树林湿地:分布、种类组成及其保护[J].亚热带植物科学,2017,46(4):301-310.
[2]杨力龙,梁芳,张小军,等.红树植物老鼠簕的研究进展[J].宁夏农林科技,2014,55(5):37-38.
[3]杨维,夏杏洲,韩维栋,等.红树植物的化学成分及其生物活性研究进展[J].食品研究与开发,2011,32(1):173-180.
[4]卢顺玉,祁平,梁颖娥,等.老鼠簕生物碱A及其衍生物对四氯化碳致大鼠肝纤维化的保护作用[J].时珍国医国药,2013,24(2):310-311.
[5]黄燕军,梅燕,祁平,等.4-羟基苯并恶唑-2-酮对四氯化碳诱导小鼠急性肝损伤的保护作用[J].医学理论与实践,2013,26(15):1961-1962,1975.
[6]Woolbright B L,Jaeschke H.Role of the inflammasome in acetaminophen-induced liver injury and acute liver failure[J].J Hepatol,2016,66(4):836-848.
[7]赵慧,王洪礼,吕佳,等.益甘宁颗粒对对乙酰氨基酚诱导小鼠肝损伤模型的保护作用[J].中南药学,2017,15(5):587-590.
[8]祁平,樊惠,梁颖娥,等.4-羟基苯并恶唑-2-酮对小鼠急性肝损伤的保护作用[J].时珍国医国药,2012,23(7):1668-1669.
[9]高雅,王刚,杜沛霖,等.白马骨水提物对急性肝损伤小鼠氧化应激及炎症反应的影响[J].中国实验方剂学杂志,2017,23(21):135-140.
[10]王巍,许立拔,张卓,等.金草消毒颗粒对D-GalN/LPS致小鼠急性肝损伤保护的影响[J].中国实验方剂学杂志,2018,24(7):108-113..
[11]汪蕾,Rahmadini N,高天慧,等.印尼姜黄对刀豆球蛋白A所致小鼠肝损伤的保护作用及其影响TGF-β1/Smad通路的机制探讨[J].中国实验方剂学杂志,2017,23(7):127-133.
[12]何银,刘春雨,何迟迟,等.扶正养肝合剂对药物性肝损伤的保护作用研究[J].中国中药杂志,2018,43(23):4685-4691.
[13]邱炳勋,刘珂,邹利,等.五味子诱导的CYPs和Nrf2活化对对乙酰氨基酚急性肝损伤的影响[J].中国中药杂志,2018,43(24):4908-4915.
[14]ZHANG Y Q,DING N,ZENG Y F,et al.New progress in roles of nitric oxide during hepatic ischemia reperfusion injury[J].World J Gastroenterol,2017,23(14),2505-2510.
[15]Mosedale M,Watkins P.Drug-induced liver injury:advances in mechanistic understanding that will inform risk management[J].Clin Pharmacol Ther,2017,doi:10.1002/cpt.564.
[16]Diesen D L,Kuo P C.Nitric oxide and redox regulation in the liver:partⅠ.general considerations and redox biology in hepatitis[J].J Surg Res,2010,162(1):95-109.
[17]Diesen D L,Kuo P C.Nitric oxide and redox regulation in the liver:partⅡ.redox biology in pathologic hepatocytes and implications for intervention[J].J Surg Res,2011,167(1):96-112.
[18]FENG J,ZHANG Q,MO W,et al.Salidroside,pretreatment attenuates apoptosis and autophagy during hepatic ischemia-reperfusion injury by inhibiting the mitogen-activated protein kinase pathway in mice[J].Drug Des Devel Ther,2017,11:1989-2006.
[19]Nakagawa H,Maeda S.Molecular mechanisms of liver injury and hepatocarcinogenesis:focusing on the role of stress-activated MAPK[J].Patholog Res Int,2012,doi:10.1155/2012/172894.
[20]ZHANG J,MIN R W,LE K,et al.The role of MAP2kinases and p38 kinase in acute murine liver injury models[J].Cell Death Dis,2017,8(6):e2903.
[21]WU L,ZHANG Q,DAI W,et al.Quercetin pretreatment attenuates hepatic ischemia reperfusion-induced apoptosis and autophagy by inhibiting ERK/NF-κBpathway[J].Gastroenterol Res Pract,2017,2:e9724217.
[2]杨力龙,梁芳,张小军,等.红树植物老鼠簕的研究进展[J].宁夏农林科技,2014,55(5):37-38.
[3]杨维,夏杏洲,韩维栋,等.红树植物的化学成分及其生物活性研究进展[J].食品研究与开发,2011,32(1):173-180.
[4]卢顺玉,祁平,梁颖娥,等.老鼠簕生物碱A及其衍生物对四氯化碳致大鼠肝纤维化的保护作用[J].时珍国医国药,2013,24(2):310-311.
[5]黄燕军,梅燕,祁平,等.4-羟基苯并恶唑-2-酮对四氯化碳诱导小鼠急性肝损伤的保护作用[J].医学理论与实践,2013,26(15):1961-1962,1975.
[6]Woolbright B L,Jaeschke H.Role of the inflammasome in acetaminophen-induced liver injury and acute liver failure[J].J Hepatol,2016,66(4):836-848.
[7]赵慧,王洪礼,吕佳,等.益甘宁颗粒对对乙酰氨基酚诱导小鼠肝损伤模型的保护作用[J].中南药学,2017,15(5):587-590.
[8]祁平,樊惠,梁颖娥,等.4-羟基苯并恶唑-2-酮对小鼠急性肝损伤的保护作用[J].时珍国医国药,2012,23(7):1668-1669.
[9]高雅,王刚,杜沛霖,等.白马骨水提物对急性肝损伤小鼠氧化应激及炎症反应的影响[J].中国实验方剂学杂志,2017,23(21):135-140.
[10]王巍,许立拔,张卓,等.金草消毒颗粒对D-GalN/LPS致小鼠急性肝损伤保护的影响[J].中国实验方剂学杂志,2018,24(7):108-113..
[11]汪蕾,Rahmadini N,高天慧,等.印尼姜黄对刀豆球蛋白A所致小鼠肝损伤的保护作用及其影响TGF-β1/Smad通路的机制探讨[J].中国实验方剂学杂志,2017,23(7):127-133.
[12]何银,刘春雨,何迟迟,等.扶正养肝合剂对药物性肝损伤的保护作用研究[J].中国中药杂志,2018,43(23):4685-4691.
[13]邱炳勋,刘珂,邹利,等.五味子诱导的CYPs和Nrf2活化对对乙酰氨基酚急性肝损伤的影响[J].中国中药杂志,2018,43(24):4908-4915.
[14]ZHANG Y Q,DING N,ZENG Y F,et al.New progress in roles of nitric oxide during hepatic ischemia reperfusion injury[J].World J Gastroenterol,2017,23(14),2505-2510.
[15]Mosedale M,Watkins P.Drug-induced liver injury:advances in mechanistic understanding that will inform risk management[J].Clin Pharmacol Ther,2017,doi:10.1002/cpt.564.
[16]Diesen D L,Kuo P C.Nitric oxide and redox regulation in the liver:partⅠ.general considerations and redox biology in hepatitis[J].J Surg Res,2010,162(1):95-109.
[17]Diesen D L,Kuo P C.Nitric oxide and redox regulation in the liver:partⅡ.redox biology in pathologic hepatocytes and implications for intervention[J].J Surg Res,2011,167(1):96-112.
[18]FENG J,ZHANG Q,MO W,et al.Salidroside,pretreatment attenuates apoptosis and autophagy during hepatic ischemia-reperfusion injury by inhibiting the mitogen-activated protein kinase pathway in mice[J].Drug Des Devel Ther,2017,11:1989-2006.
[19]Nakagawa H,Maeda S.Molecular mechanisms of liver injury and hepatocarcinogenesis:focusing on the role of stress-activated MAPK[J].Patholog Res Int,2012,doi:10.1155/2012/172894.
[20]ZHANG J,MIN R W,LE K,et al.The role of MAP2kinases and p38 kinase in acute murine liver injury models[J].Cell Death Dis,2017,8(6):e2903.
[21]WU L,ZHANG Q,DAI W,et al.Quercetin pretreatment attenuates hepatic ischemia reperfusion-induced apoptosis and autophagy by inhibiting ERK/NF-κBpathway[J].Gastroenterol Res Pract,2017,2:e9724217.