重度创伤性脑损伤致应激性肝损伤大鼠肝组织细胞色素P4502El水平变化

doi:10.3969/j.issn.1672-5069.2014.05.017

摘要/Abstract

摘要： 目的研究重度创伤性脑损伤（TBI）诱导的应激性肝损害（HSI）大鼠肝组织细胞色素P4502El（CYP2E1）的变化。方法取40只健康雄性Wistar大鼠,随机分为假手术(A组)和TBI组,采用改良的Feeney自由落体撞击法建立TBI大鼠模型;于颅脑致伤后6、12和24 h,检测各组血清ATL、AST水平和丙二醛（MDA）水平变化,在光镜下观察肝脏组织学改变,采用RT-PCR和Western blot法分别检测各组肝组织CYP2E1 mRNA和蛋白表达。结果在TBI后6和24 h,各组大鼠血清ALT较基线水平[（42.2±8.1） U/L]明显升高[分别为（83.0±10.3） U/L 和（1204.5±146.6） U/L,P<0.01）];伤后12 h血清AST较基线[（44.0±7.2） U/L]升高[（280.4±53.3） U/L,P<0.01）],于24 h达峰值[（790.3±114.5） U/L];伤后6 h MDA较基线[（5.2±0.2） nmol/mg]明显升高[（14.2±0.2） nmol/mg,P<0.05],24 h达峰值[（56.3±0.5） nmol/mg];伤后24 h肝组织损伤最严重,可见肝小叶结构不清,肝窦明显扩张,散在肝细胞点状坏死,大量炎细胞浸润;伤后6 h肝组织CYP2E1 mRNA和蛋白表达水平较基线水平[分别为（0.28±0.02）和（61.68±0.60）]明显增加[（0.89±0.05）和（120.24±1.22）,P<0.05],在24 h达峰值[分别为（1.50±0.02）和（200.40±2.61）,P<0.01]。结论CYP2E1可能参与了TBI诱导的氧化应激反应,从而引起HSI。

关键词: 肝损伤, 应激反应, 细胞色素P4502El, 氧化应激, 大鼠

Abstract: Objective To explore the dynamic changes of cytochrome P4502El（CYP2E1）in liver tissues of rats with hepatic stress injury（HSI） secondary to traumatic brain injury（TBI）. Methods Forty healthy male Wistar rats were randomly divided into sham-operated and TBI group. The animal model was established by an improved Feeney method. Serum levels of ALT and AST were measured by enzymatic assay at 6,12 and 24 h after TBI;MDA contents in liver tissues were also measured. Pathological changes of liver tissues were observed under light microscopy. The CYP2E1 mRNA levels and its protein expression were detected by RT-PCR and Western blot,respectively. Results At 6 h and 24 h after TBI,serum ALT elevated significantly [（83.0±10.3） U/L and（1204.5±146.6） U/L,respectively,P<0.01）] as compared with baseline [（42.2±8.1） U/L];at 12 h after TBI,serum AST elevated significantly[（280.4±53.3） U/L,P<0.01）] compared with baseline[（44.0±7.2）） U/L],and it peaked at 24 h[（790.3±114.5） U/L];at 6 h after TBI,serum MDA elevated significantly [（14.2±0.2） nmol/mg,P<0.05] compared with baseline[（5.2±0.2） nmol/mg],and it peaked at 24 h after TBI [（56.3±0.5）nmol/mg];at 24 h after TBI,the injuries in liver tissues were serious,with expanded sinus,scattered spotty necrosis and inflammatory cell infiltration;at 6 h after TBI,both mRNA and protein levels of CYP2E1 were significantly elevated[（0.89±0.05）and（120.24±1.22）,P<0.05] compared with baseline[（0.28±0.02） and（61.68±0.60）,respectively],and they peaked at 24 h after TBI[（1.50±0.02）and（200.40±2.61）,respectively,P<0.01]. ConclusionsCYP2E1 may be involved in oxidative stress in hepatic stress injury after TBI.

Key words: Hepatic stress injury, Cytochrome P4502El, Oxidative stress, Rats

李静, 李毅, 刘天会, 尤红, 徐有青. 重度创伤性脑损伤致应激性肝损伤大鼠肝组织细胞色素P4502El水平变化[J]. 实用肝脏病杂志, 2014, 17(5): 515-518.

Li Jing, Li Yi, Liu Tianhui. Dynamic changes of cytochrome P4502El in liver tissues of rats with hepatic stress injury secondary to traumatic brain injury[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2014, 17(5): 515-518.

参考文献

[1] 孟庆颖,朱玉群. 成人创伤性脑损伤后早期应激性肝损害的临床分析. 首都医科大学学报,2007,24（4）:93-95.
[2] Zygun DA,Kortbeek JB,Fick GH,et al. Non-neurologic organ dysfunction in severe traumatic brain injury.Crit Care Med,2005,33（3）:654-60.
[3] Fang CW,Yao YM,Zhai HX,et al. Tissue lipopolysaccharide-binding protein expression in rats after thermal injury: potential role of TNF-alpha. Burns,2004,30（3）:225-231.
[4] Raza H. Dual localization of glutathione S-transferase in the cytosol and mitochondria:Implications in oxidative stress,toxicity and disease. FEBS J,2011,278（22）:4243-4251.
[5] Wanders RJ,Ferdinandusse S,Brites P,et al. Peroxisomes,lipid metabolism and lipot oxicity.Biochim Biophys Acta,2010,1801（3）:272-280.
[6] Abdelmegeed MA,Moon KH,Chen C,et al. Role of cytochrome P450 2E1 in protein nitration and ubiquitin-mediated degradation during acetaminophen toxicity. Biochem Pharmacol,2010,79（1）:57-66.
[7] Gong WH,Zheng WX,Wang J,et al. Coexistence of hyperlipidemia and acute cerebral ischemia/reperfusion induces severe liver damage in a rat model. World J Gastroenterol,2012,18（35）:4934-4943.
[8] Schoch KM,Madathil SK,Saatman KE. Genetic manipulation of cell death and neuroplasticity pathways in traumatic brain injury. Neurotherapeutics,2012,9（2）:323-337.
[9] Begriche K,Igoudjil A,Pessayre D,et al. Mitochondrial dysfunction in NASH: causes,consequences and possible means to prevent it. Mitochondrion,2006,6（1）:1-28.
[10] Song BJ,Moon KH,Upreti VV,et al. Mechanisms of MDMA （Ecstasy）-induced oxidative stress,mitochondrial dysfunction,and organ damage. Curr Pharm Biotechnol,2010,11（5）:434-443.
[11] Schreuder TC,Verwer BJ,van Nieuwkerk CM,et al. Nonalcoholic fatty liver disease:an overview of current insights in pathogenesis,diagnosis and treatment.World J Gastroenterol,2008,14（16）:2474-2486.
[12] Lu Y,Cederbaum AI. CYP2E1 and oxidative liver injury by alcohol. Free Radic Biol Med,2008,44（5）:723-738.
[13] Morris EM,Rector RS,Thyfault JP,et al. Mitochondria and redox signaling in steatohepatitis. Antioxid Redox Signal,2011,15（2）:485-504.
[14] Reed JR,Cawley GF,Backes WL. Inhibition of cytochrome P450 1A2-mediated metabolism and production of reactive oxygen species by heme oxygenase-1 in rat liver microsomes.Drug Metab Lett,2011,5（1）:6-16.
[15] Liu J,Litt L,Segal MR,et al. Metabolomics of oxidative stress in recent studies of endogenous and exogenously administered intermediate metabolites.Int J Mol Sci,2011,12（10）:6469-6501.
[16] Gonzales S,Polizio AH,Erario MA,et al. Glutamine is highly effective in preventing in vivo cobalt-induced oxidative stress in rat liver. World J Gastroenterol,2005,11（23）:3533-3538.
[17] Daly AK. Relevance of CYP2E1 to non-alcoholic fatty Liver Disease. Subcell Biochem,2013,67:165-175.
[18] Cederbaum AI. Role of CYP2E1 in ethanol-induced oxidant stress,fatty liver and hepatotoxicity. Dig Dis,2010,28（6）:802-811.
[19] Cederbaum AI,Lu Y,Wu D. Role of oxidative stress in alcohol-induced liver injury. Arch Toxicol,2009,83（6）:519-548.
[20] Lu Y,Cederbaum AI. CYP2E1 potentiation of LPS and TNFα-induced hepatotoxicity by mechanisms involving enhanced oxidative and nitrosative stress,activation of MAP kinases, and mitochondrial dysfunction.Genes Nutr,2010,5（2）:149-167.
[21] 梅玫,陆伟. 细胞色素P450 2E1在脂肪性肝病中致病作用研究进展. 实用肝脏病杂志,2007,10（1）:74-76.
[22] Yang L,Wu D,Wang X,et al. Cytochrome P4502E1, oxidative stress,JNK,and autophagy in acute alcohol-induced fatty liver. Free Radic Biol Med,2012,53（5）:1170-1180.