CCl_4致大鼠肝损伤24 h生物标志物水平的变化规律
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摘要
目的:采用四氯化碳(CCl_4)复制大鼠肝损伤的动物模型,研究肝损伤早期生物标志物谷氨酸脱氢酶(GLDH),嘌呤核苷酸磷酸化酶(PNP),α-谷胱甘肽-S-转移酶(α-GST),精氨酸酶1(Arg1)的动态变化,为急性肝损伤的早期发现提供实验依据。方法:48只Wistar大鼠,随机分为正常组和模型组,模型组腹腔注射10%的CCl_4橄榄油溶液10 m L·kg~(-1),禁食不禁水,于3,6,12,24 h后处死各组动物,测定血清肝功能丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST),胆红素(TBIL),碱性磷酸酶(ALP)的水平,α-GST,Arg1,GLDH,PNP水平,肝匀浆超氧化物歧化酶(SOD),谷胱甘肽(GSH),丙二醛(MDA)的水平。结果:与正常组比较,给药3 h后ALT,AST,TBIL,α-GST,Arg1,GLDH,PNP,MDA水平显著升高,SOD明显下降(P <0. 01),给药6,12 h后ALT,AST,α-GST,Arg1,GLDH,TBIL,ALP,MDA水平显著升高,GSH,SOD明显下降(P <0. 05,P <0. 01),给药24 h后ALT,AST,α-GST,Arg1,TBIL,ALP,MDA水平显著升高,GSH,SOD显著下降(P <0. 01)。结论:α-GST,Arg1,GLDH,PNP较传统肝功能检测指标具有较好的灵敏性,可用于CCl_4致大鼠肝损伤早期检测。
Objective: To replicate the animal model of liver injury in rats by using carbon tetrachloride( CCl_4),investigate the dynamic changes of early biomarkers of liver injury,namely glutamate dehydrogenase( GLDH),purine nucleotide phosphorylase( PNP),α-dynamic changes of glutathione-S-transferase( α-GST) and arginase 1( Arg1),and provide experimental evidence for early detection of acute liver injury. Method: Fortyeight Wistar rats were randomly divided into a blank group and a model group. The model group was intraperitoneally injected with 10 m L·kg-110% CCl_4 olive oil solution,fasting but except water. Animals were sacrificed at 3,6,12, and 24 h. The serum liver function alanine aminotransferase( ALT), aspartate aminotransferase( AST),bilirubin( TBIL),alkaline phosphatase( ALP) levels,α-GST,Arg1,GLDH,PNP levels,and liver homogenate superoxide dismutase( SOD),glutathione( GSH),malondialdehyde( MDA) levels were then detected. Result: As compared with blank group,the levels of ALT,AST,TBIL,α-GST,Arg1,GLDH,PNP and MDA were increased significantly 3 h after administration,and SOD was decreased significantly( P < 0. 01). After 6,12 h,the levels of ALT,AST,α-GST,ARG-,GLDH,TBIL,ALP and MDA were increased significantly,while GSH and SOD were decreased significantly( P < 0. 05,P < 0. 01). After 24 h,the levels of ALT,AST,α-GST,Arg1,TBIL,ALP and MDA were significantly increased,while GSH and SOD were significantly decreased( P < 0. 01). Conclusion: α-GST,Arg1,GLDH and PNP have better sensitivity than traditional liver function test indicators,and can be used for early detection of liver injury induced by CCl_4 in rats.
Objective: To replicate the animal model of liver injury in rats by using carbon tetrachloride( CCl_4),investigate the dynamic changes of early biomarkers of liver injury,namely glutamate dehydrogenase( GLDH),purine nucleotide phosphorylase( PNP),α-dynamic changes of glutathione-S-transferase( α-GST) and arginase 1( Arg1),and provide experimental evidence for early detection of acute liver injury. Method: Fortyeight Wistar rats were randomly divided into a blank group and a model group. The model group was intraperitoneally injected with 10 m L·kg-110% CCl_4 olive oil solution,fasting but except water. Animals were sacrificed at 3,6,12, and 24 h. The serum liver function alanine aminotransferase( ALT), aspartate aminotransferase( AST),bilirubin( TBIL),alkaline phosphatase( ALP) levels,α-GST,Arg1,GLDH,PNP levels,and liver homogenate superoxide dismutase( SOD),glutathione( GSH),malondialdehyde( MDA) levels were then detected. Result: As compared with blank group,the levels of ALT,AST,TBIL,α-GST,Arg1,GLDH,PNP and MDA were increased significantly 3 h after administration,and SOD was decreased significantly( P < 0. 01). After 6,12 h,the levels of ALT,AST,α-GST,ARG-,GLDH,TBIL,ALP and MDA were increased significantly,while GSH and SOD were decreased significantly( P < 0. 05,P < 0. 01). After 24 h,the levels of ALT,AST,α-GST,Arg1,TBIL,ALP and MDA were significantly increased,while GSH and SOD were significantly decreased( P < 0. 01). Conclusion: α-GST,Arg1,GLDH and PNP have better sensitivity than traditional liver function test indicators,and can be used for early detection of liver injury induced by CCl_4 in rats.
引文
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[15]邱平,唐明薇,梅希,等.病理学诊断的非酒精性脂肪性肝病患者的临床特点分析[J].川北医学院学报,2015,30(2):182-186,194.
[16]刘晓娜,彭双清,贾栗,等.对乙酰氨基酚致小鼠肝损伤早期血清苹果酸脱氢酶和嘌呤核苷磷酸化酶的变化[J].毒理学杂志,2015,29(1):54-56.
[17]贺蕾艳,郭瑶雪,李春,等.药物性肝损伤生物标志物研究进展[J].药学学报,2015,50(8):959-965.
[18] Schomaker S,Warner R,Bock J,et al. Assessment of emeRGing biomarkers of liver injury in human subjects[J]. Toxicol Sci,2013,132(2):276-283.
[19] Aubrecht J, Schomaker S. Serum glutamate dehydrogenase as a potential biomarker of mitochondrial dysfunction[J]. Toxicol Sci,2013,134(1):223.
[20] Thulin P,Hornby R J,Auli M,et al. A longitudinal assessment of miR-122 and GLDH as biomarkers of drug-induced liver injury in the rat[J]. Biomarkers,2017,22(5):461-469.
[21]乔靖怡,周璐,金若敏,等.四氯化碳致大鼠肝损伤早期血清总胆汁酸、α-谷胱甘肽S转移酶、嘌呤核苷磷酸化酶和鸟氨酸氨基甲酰转移酶的变化[J].中国药理学与毒理学杂志,2013,27(4):650-656.
[2] Amacher D E,Schomaker S J,Aubrecht J,et al.Development of blood biomarkers for drug induced liver injury:an evaluation of their potential for risk assessment and diagnostic[J]. Mol Diagn Ther,2013,17(6):343-354.
[3]马蓉,李茂星,贾正平,等.螃蟹甲大孔吸附树脂50%乙醇洗脱物对大鼠CCl4肝损伤的保护作用[J].中国实验方剂学杂志,2016,22(4):132-136.
[4] Almazroo O A,Miah M K,Venkataramanan R. Drug metabolism in the liver[J]. Clin Liver Dis,2017,21(1):1-20.
[5]乔靖怡,白明,苗明三.基于肝病临床病症特点的动物模型分析[J].中华中医药杂志,2017,32(2):582-587.
[6] Abbas A T,El-Shitany N A,Shaala L A,et al. Red sea subereamollis sponge extract protects against CCl4-induced acute liver injuryin ratsviaan antioxidant mechanism[J]. Evid-Based Complement Alternat Med,2014,doi:org/10. 1155. 2014/745606.
[7] Abdel-moneim A M,Al-Kahtani M A,El-Kersh M A,et al. Free radical-scavenging, anti-inflammatory/antifibrotic and hepatoprotective actionsof taurine and silymarin against CCl4induced rat liver damage[J].PLo S One,2015,10(12):1-16.
[8] Rosillo M A,Sanchez-Hidalgo M,Cárdeno A,et al.Protective effect of ellagic acid,a natural polyphenolic compound,in a murine model of Crohn's disease[J].Biochem Pharmacol,2011,82(7):737-745.
[9] CHEN X,GONG X,JIANG R,et al. Resolvin D1attenuates CCl4-induced acute liver injury involving upregulation of HO-1 in mice[J]. Immunopharmacol Immunotoxicol,2016(2):61-67.
[10] Shabaneh A 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.
[11]周健.总胆红素与代谢综合征及其诊断组分的关系研究[J].中国实验诊断学,2018,22(3):422-425.
[12] Maeda K. ORGanic anion transporting polypeptide(OATP)1B1 and OATP1B3 as important regulators of the pharmacokinetics of substrate drugs[J]. Biol Pharm Bull,2015,38(2):155-168.
[13]邵坚,罗光明,朱继孝,等.栀子不同炮制饮片对四氯化碳致急性肝损伤大鼠的保护作用[J].中药材,2016,39(7):1521-1524.
[14]王跃峰,张可锋,周雨晴,等.拳卷地钱总黄酮对四氯化碳致急性肝损伤大鼠的保护作用及其作用机制[J].时珍国医国药,2017,28(2):277-279.
[15]邱平,唐明薇,梅希,等.病理学诊断的非酒精性脂肪性肝病患者的临床特点分析[J].川北医学院学报,2015,30(2):182-186,194.
[16]刘晓娜,彭双清,贾栗,等.对乙酰氨基酚致小鼠肝损伤早期血清苹果酸脱氢酶和嘌呤核苷磷酸化酶的变化[J].毒理学杂志,2015,29(1):54-56.
[17]贺蕾艳,郭瑶雪,李春,等.药物性肝损伤生物标志物研究进展[J].药学学报,2015,50(8):959-965.
[18] Schomaker S,Warner R,Bock J,et al. Assessment of emeRGing biomarkers of liver injury in human subjects[J]. Toxicol Sci,2013,132(2):276-283.
[19] Aubrecht J, Schomaker S. Serum glutamate dehydrogenase as a potential biomarker of mitochondrial dysfunction[J]. Toxicol Sci,2013,134(1):223.
[20] Thulin P,Hornby R J,Auli M,et al. A longitudinal assessment of miR-122 and GLDH as biomarkers of drug-induced liver injury in the rat[J]. Biomarkers,2017,22(5):461-469.
[21]乔靖怡,周璐,金若敏,等.四氯化碳致大鼠肝损伤早期血清总胆汁酸、α-谷胱甘肽S转移酶、嘌呤核苷磷酸化酶和鸟氨酸氨基甲酰转移酶的变化[J].中国药理学与毒理学杂志,2013,27(4):650-656.