己酮可可碱对小鼠酒精性肝病酒精代谢酶和核受体PPAR-α的影响

doi:10.3969/j.issn.1672-5069.2014.02.013

摘要/Abstract

摘要： 目的研究己酮可可碱（PTX）对酒精性肝病小鼠酒精代谢酶和过氧化物酶增殖物激活受体（PPAR-α）的影响。方法将64只C57BL/6小鼠随机分为模型组、治疗组和对照组，用50%酒精灌胃建立急性肝损伤模型，以20%酒精连续灌胃6周建立慢性酒精性肝病模型；采用比色法检测各组小鼠血清乙醇脱氢酶（ADH）和细胞色素P4502E1（CYP2E1)活性；采用RT-PCR法检测肝组织ADH、CYP2E1和PPAR-α mRNA水平；采用免疫组化法检测肝组织CYP2E1和PPAR-α蛋白表达。结果急性和慢性酒精性肝损伤模型小鼠血清ADH活性分别为（11.2±1.6）U/ml和（5.8±1.4）U/ml，与相应对照组比无显著性差异[分别为（12.5±1.2）U/ml和（4.3±0.6）U/ml]；急性和慢性酒精性肝损伤小鼠CYP2E1活性分别为（12.2±1.8）U/ml和（11.8±1.7）U/ml，均显著高于对照组[（7.9±1.4）U/ml和（6.5±1.2）U/ml，P<0.01）]和治疗组[（8.1±1.5）U/ml和（7.8±1.5）U/ml，P<0.01]；急性和慢性酒精性肝损伤小鼠肝组织CYP2E1阳性细胞相对表达强度为（765±21）和（682±25），均显著高于对照组[分别为（308±12）和（305±18），P<0.01）]和大剂量PTX治疗组[分别为（521±18）和（418±12），P<0.01]；急性和慢性酒精性肝损伤小鼠肝组织ADH mRNA水平与对照组比无显著性差异，但肝组织CYP2E1 mRNA相对水平分别为（1.47±0.32）和（1.13±0.52），显著高于对照组[（0.89±0.23）和（0.45±0.28），P<0.01）]及大剂量PTX治疗组[分别为（0.92±0.27）和（0.48±0.32），P<0.01）]；急性酒精性肝病动物肝组织PPAR-α mRNA水平与对照组或PTX治疗组比无统计学差异，但慢性酒精性肝损伤小鼠肝组织PPAR-α mRNA相对水平[（0.45±0.31）]显著低于对照组[（0.85±0.21），（P<0.05）]；急性和慢性酒精性肝损伤小鼠肝组织PPAR-α阳性相对表达强度为（322±15）和（262±23），均显著低于对照组[分别为（721±18）和（689±14），（P<0.01）]和大剂量PTX治疗组[分别为（548±20）和（725±19），P<0.01）]。结论 PTX能够减轻急慢性酒精性肝损伤，可能与其上调酒精代谢酶CYP2E1和下调PPAR-α表达有关，而与ADH无关。

关键词: 酒精性肝病, 酒精代谢酶, 己酮可可碱, 过氧化物酶增殖物激活受-α, 小鼠

Abstract: Objective To investigate the effect of pentoxifylline （PTX） on ethanol metabolic enzymes and peroxisome proliferator-activated receptor alpha （PPAR-α） in C57BL/6 mice with alcoholic liver disease. Methods Sixty-four mice were randomly divided into alcoholic liver disease model， PTX intervention and control group; Acute alcoholic liver injury was induced in mice by intragastric administration with 50% alcohol，and chronic alcoholic liver injury was induced by intragastric administration with 20% alcohol daily for six weeks；Serum alcohol dehydrogenase（ADH） and cytochrome P4502E1 （CYP2E1） activity was measured by colorimetric method；The mRNA levels of ADH，CYP2E1 and PPAR-α were measured by PT-PCR；The protein expression of CYP2E1 and PPAR-α was determined by immunohistochemistry. Results The serum activity of ADH did not differ among mice with acute[（11.2±1.6）U/ml] or chronic[（5.8±1.4） U/ml] alcoholic liver injury and controls[（12.5±1.2）U/ml and（4.3±0.6）U/ml，respectively]；In mice with acute and chronic liver injury，the CYP2E1 activity was（12.2±1.8）U/ml and （11.8±1.7） U/ml，respectively，significantly higher than those in normal controls[（7.9±1.4）U/ml and （6.5±1.2） U/ml，P<0.01）] and those in PTX-intervented mice [（8.1±1.5） U/ml and （7.8±1.5）U/ml，P<0.01]；The relative CYP2E1 positive cells in liver tissues in mice with acute and chronic alcoholic liver injury were （765±21） and （682±25），respectively，significantly higher than those in normal controls [（308±12） and （305±18），P<0.01）] and in mice with high-dosage of PTX intervention [（521±18） and （418±12），respectively，P<0.01]；The mRNA levels of ADH in liver tissues of mice with acute and chronic alcoholic liver injury were similar to that in the controls， however， the relative CYP2E1 mRNA levels[（1.47±0.32） and （1.13±0.52）] were significantly higher than those in normal controls [（0.89±0.23） and （0.45±0.28），respectively，P<0.01）] and in mice with high-dose of PTX[（0.92±0.27） and （0.48±0.32），respectively，P<0.01）];PPAR-α mRNA levels in liver tissues did not differ among mice with acute liver injury and normal controls，however，it was significantly lower than that in normal controls（0.85±0.21） in liver of mice with chronic alcoholic liver injury[（0.45±0.31），P<0.05]；The PPAR-α positive cells in mice of acute and chronic alcohol injury were （322±15） and（262±23），respectively，significantly higher than those in normal controls[（721±18）and （689±14），P<0.01] and in mice with high-dose of PTX intervention[（548±20） and （725±19），P<0.01]. Conclusion PTX attenuates acute or chronic alcoholic liver injury，probably by through up-regulation of CYP2E1 and down-regulation of PPAR-α.

Key words: Alcoholic liver disease, Ethanol metabolic enzymes, Pentoxifylline, Peroxisome proliferator-activated receptor alpha, Mice

屈耀宁，董蕾，史海涛，秦斌，刘亚萍. 己酮可可碱对小鼠酒精性肝病酒精代谢酶和核受体PPAR-α的影响[J]. 实用肝脏病杂志, 2014, 17(2): 163-167.

Qu Yaoning，Dong Lei，Shi Haitao，et al.. Effects of pentoxifylline on ethanol metabolic enzymes and peroxisome proliferator-activated receptor alpha in mice with alcoholic liver disease[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2014, 17(2): 163-167.

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