酒精致大鼠肝损伤早期血清生物标志物水平的变化规律
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摘要
目的:研究酒精性肝损伤的生物标志物谷氨酸脱氢酶(GLDH),α-谷胱甘肽-S-转移酶(α-GST),嘌呤核苷酸磷酸化酶(PNP),精氨酸酶1(Arg1)的动态变化,明确这些指标联合检测是否可以作为酒精性肝损伤早诊断生物标志物。方法:Wistar大鼠48只,随机分为正常组和模型组2组,每组24只,雌雄各半。每日禁食但不禁水7 h后,模型组灌胃50%乙醇/10 mg·kg~(-1),正常组灌服同样体积的生理盐水; 1 h后按前用量重复灌胃50%乙醇1次,正常组灌服同体积生理盐水,连续造模和给药6 d,造成急性酒精性肝损伤模型。在第2,3,4,6天末次酒精灌胃3 h后,分别随机取2组大鼠每组6只雌雄各半,共12只,处死各组动物,测定天门冬氨酸氨基转移酶(AST),丙氨酸氨基转移酶(ALT),碱性磷酸酶(ALP),胆红素(TBIL),GLDH,α-GST,PNP,Arg1水平。结果:与正常组比较,模型组大鼠ALT,AST,ALP,TBIL,GLDH,PNP,α-GST,Arg1水平差异显著(P <0. 01),表明酒精性肝损伤模型已成功建立。模型组大鼠GLDH,PNP,α-GST,Arg1比ALT,AST水平升高时间早,幅度大。结论:GLDH,PNP,α-GST,Arg1较传统肝功能检测有较强的灵敏性,可以作为酒精性肝损伤早期检测生物标志物。
Objective: To investigate the dynamic changes of the biomarkers of alcoholic liver injury,including glutamate dehydrogenase( GLDH), α-glutathione-S-transferase( α-GST), purine nucleotide phosphorylase( PNP),and arginine enzyme 1( Arg1),and clarify whether these indexes can be used as early diagnostic biomarkers for alcoholic liver injury. Method: 48 Wistar rats were randomly divided into a blank group and a model group,24 rats in each group,half male and half female. After fasting but except water for 7 h,50% ethanol/10 m L·kg~(-1) was given to the model group by intragastric administration and the same volume of normal saline was administered to the blank group. After 1 h,50% ethanol was again given for once by intragastric administration according to the previous dosage. In the blank group,the same volume of normal saline was administered. After modeling and administration for 6 d,acute alcoholic liver injury model was established. 3 h after the last intragastric administration of alcohol at day 2,3,4,6,six rats( half male and half female) in each group were randomly selected. All the animals were sacrificed to determine the aspartate aminotransferase( AST),alanine aminotransferase( ALT),alkaline phosphatase( ALP),bilirubin( TBIL),GLDH,α-GST,PNP,and Arg1 levels. Result: As compared with the blank group,the levels of ALT,AST,ALP,TBIL,GLDH,PNP,α-GST and Arg1 in the model group were significantly different( P < 0. 01),indicating that the alcoholic liver injury model was successfully established. In the model group,GLDH,PNP,α-GST and Arg1 levels were increased earlier and more significantly than ALT and AST levels. Conclusion: GLDH,PNP,α-GST and Arg1 can be used as biomarkers for early detection of alcoholic liver injury.
Objective: To investigate the dynamic changes of the biomarkers of alcoholic liver injury,including glutamate dehydrogenase( GLDH), α-glutathione-S-transferase( α-GST), purine nucleotide phosphorylase( PNP),and arginine enzyme 1( Arg1),and clarify whether these indexes can be used as early diagnostic biomarkers for alcoholic liver injury. Method: 48 Wistar rats were randomly divided into a blank group and a model group,24 rats in each group,half male and half female. After fasting but except water for 7 h,50% ethanol/10 m L·kg~(-1) was given to the model group by intragastric administration and the same volume of normal saline was administered to the blank group. After 1 h,50% ethanol was again given for once by intragastric administration according to the previous dosage. In the blank group,the same volume of normal saline was administered. After modeling and administration for 6 d,acute alcoholic liver injury model was established. 3 h after the last intragastric administration of alcohol at day 2,3,4,6,six rats( half male and half female) in each group were randomly selected. All the animals were sacrificed to determine the aspartate aminotransferase( AST),alanine aminotransferase( ALT),alkaline phosphatase( ALP),bilirubin( TBIL),GLDH,α-GST,PNP,and Arg1 levels. Result: As compared with the blank group,the levels of ALT,AST,ALP,TBIL,GLDH,PNP,α-GST and Arg1 in the model group were significantly different( P < 0. 01),indicating that the alcoholic liver injury model was successfully established. In the model group,GLDH,PNP,α-GST and Arg1 levels were increased earlier and more significantly than ALT and AST levels. Conclusion: GLDH,PNP,α-GST and Arg1 can be used as biomarkers for early detection of alcoholic liver injury.
引文
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[18]曾瑾,唐绍微,刘云华,等.苍耳子对正常大鼠重复给药的肝毒性效应及其机制研究[J].中药药理与临床,2018,34(2):79-82.
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[20]刘晓娜,刘密凤,贾栗,等.药源性肝损伤潜在血清生物标志物的研究进展[J].首都医科大学学报,2014,35(4):511-515.
[21]中华医学会肝病学分会脂肪肝和酒精性肝病学组.酒精性肝病诊疗指南[J].中国肝脏病杂志:电子版,2010,2(4):49-53.
[22]李洪亮,张秋芳,郑雪皎,等.中药抗肝氧化应激的研究进展[J].医药导报,2013,32(12):1608-1612.
[2]王咪娜,徐士勋,林锦璇,等.鳖甲寡肽I-C-F-6对乙醇诱导的小鼠慢性肝损伤的实验研究[J].天津中医药,2015,32(1):26-29.
[3] WANG F S,FAN J G,ZHANG Z,et al. The global burden of liver disease:the major impact of China[J].Hepatology,2014,60(6):2099-2108.
[4]刘小茜,吴文晓,耿兴超,等.药物性肝损伤生物标志物研究进展[J].中国新药杂志,2018,27(1):47-52.
[5]柴玮杰,孙建飞,王明秋,等.大鼠亚急性酒精性肝损伤模型的建立[J].实验动物科学,2018,35(2):47-51.
[6]陶施民,卢贤欢,郭雅娟,等.葛根枳椇子栀子胶囊解酒护肝功效研究[J].中医药导报,2018,24(10):19-22,27.
[7] Rehm J,Samokhvalov A V,Shield K D. Global burden of alcoholic liver diseases[J]. J Hepatol,2013,59(1):160-168.
[8]吕圭源,陈素红,张丽丹,等.铁皮石斛对小鼠慢性酒精性肝损伤模型血清2种转氨酶及胆固醇的影响[J].中国实验方剂学杂志,2010,16(6):192-193.
[9]徐博,吴畏难,李传甲,等.萱草花总黄酮对小鼠急性酒精性肝损伤保护作用及机制探讨[J].中国实验方剂学杂志,2016,22(23):139-143.
[10]张艳芳,黄晶.酒精性肝纤维化的血清标志物研究进展[J].临床肝胆病杂志,2018,34(3):623-626.
[11] Shabaneh A I,Tamimi H A,McDonald R. Elevated alanine aminotransferase levels associated with polymyositis:can this be due to muscle injury?[J]. J Clin Rheumatol,2008,14(6):363-364.
[12] Amacher D E,Schomaker S J,Aubrecht J,et al.Development of blood biomarkers for drug induc; ed liver injury:an evaluation of their potential for risk assessment and diagnostic[J]. Mol Diagn Ther,2013,17(6):343-354.
[13]李飞,陆伦根.肝功能异常的评估以及相关肝病的诊断思路[J].胃肠病学,2018,23(5):305-308.
[14] Raoul Poupon. Liver alkaline phosphatase:a missing link between choleresis and biliary inflammation[J].Hepatology,2015,61(6):2080-2090.
[15] Ozer J,Ratner M,Shaw M,et al. Schomaker S. The current state of serum biomarkers of hepatotoxicity[J].Toxicology,2008,245(3):194-205.
[16] YAMAMOTO T,KIKKAWA R,YAMADA H,et al.Investigation of proteomic biomarkers in vivo hepatotoxicity study of rat liver:toxicity differentiation in hepatotoxicants[J]. J Toxicol Sci,2006,31(1):49-60.
[17]赵元明.血清谷氨酸脱氢酶活性检测在肝胆疾病中的临床价值[J].检验医学,2013,28(2):163-165.
[18]曾瑾,唐绍微,刘云华,等.苍耳子对正常大鼠重复给药的肝毒性效应及其机制研究[J].中药药理与临床,2018,34(2):79-82.
[19]耿兴超,沈连忠,李波,等.肝毒性生物标志物研究进展[J].中国药学杂志,2011,46(10):721-725.
[20]刘晓娜,刘密凤,贾栗,等.药源性肝损伤潜在血清生物标志物的研究进展[J].首都医科大学学报,2014,35(4):511-515.
[21]中华医学会肝病学分会脂肪肝和酒精性肝病学组.酒精性肝病诊疗指南[J].中国肝脏病杂志:电子版,2010,2(4):49-53.
[22]李洪亮,张秋芳,郑雪皎,等.中药抗肝氧化应激的研究进展[J].医药导报,2013,32(12):1608-1612.