利用心脏特异性CYP2E1基因修饰小鼠评价药物心脏毒性的初步探索  

Exploration of the use of heart-specific CYP2E1 genetically modified mice in evaluation of drug-induced cardiotoxicity

作　　者：裴彦宇 孙井江[1] 赵显莉 贺银丽[1] 魏若尧 高虹[1] 

机构地区：[1]中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,北京100021

出　　处：《中国实验动物学报》2016年第1期53-58,79,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：十二五重大新药创制科技项目(编号:2013ZX09302302)

摘　　要：目的使用两种心脏特异性CYP2E1基因修饰小鼠来评价药物西布曲明(sibutramine)的心脏毒性作用,选择出在评价药物的心脏毒性中具有更高敏感性的动物模型。方法 8周龄雄性Tg(+)型小鼠、s Tg(+)型小鼠、野生型C57BL/6小鼠(WT),各50只,分别随机分成5组,每组10只,溶媒对照组(灌胃给予纯净饮用水)及4个实验组(50、100、150、300 mg/kg西布曲明)。给药过程中进行一般症状观察以及存活率的测定;给药15 d后取血并解剖取心脏,测定各组实验小鼠血生化心脏损伤指标:乳酸脱氢酶(LDH),肌酸激酶(CK),肌酸激酶同工酶(CK-MB)。并进行病理组织学检查,免疫组织化学方法观察心脏组织缝隙蛋白43(connexin 43,CX43)的表达情况。结果 (1)血生化指标显示,在给药剂量50 mg/kg和100 mg/kg时,Tg(+)型小鼠均显著高于WT型小鼠(P<0.05或P<0.01);在给药剂量50、100和150 mg/kg时,Tg(+)型小鼠均显著高于s Tg(+)型小鼠(P<0.05或P<0.01);而在给药剂量300 mg/kg时,Tg(+)型小鼠均显著低于WT和s Tg(+)型小鼠(P<0.05或P<0.01),且s Tg(+)型小鼠减小的程度最小。(2)病理组织学结果显示,Tg(+)和WT型小鼠各个给药组均表现出心脏损伤的迹象。(3)免疫组织化学染色显示,随着给药剂量的加大,Tg(+)、s Tg(+)和WT型三种小鼠CX43在心肌细胞闰盘处的表达数量均逐渐下降,且染色颜色逐渐变浅;而CX43在心肌细胞闰盘处的表达数量下降程度和染色颜色变浅程度由高到低分别为,Tg(+)型小鼠,其次是WT型小鼠,s Tg(+)型小鼠。结论 Tg(+)型小鼠在评价药物潜在心脏毒性试验中可能比WT型小鼠具有更高的敏感性,而s Tg(+)型小鼠则是很好的心脏毒性保护模型。Objective To select more sensitive animal model to evaluate the cardiotoxicity of drugs,we used the heart-specific CYP2E1 genetically modified mice（ α-MHC CYP2E1 transgenic mice [Tg（ ＋） mice] and α-MHC CYP2E1 silencing transgenic mice [s Tg（ ＋） mice]） to evaluate the drug sibutramine-induced cardiotoxicity. Methods The 8-week old male Tg（ ＋）,s Tg（ ＋）,and C57 BL /6 mice（ wild type,WT）,50 mice in each group,were randomly divided into 5 groups： the solvent control group（ intragastric gavage of pure drinking water）,and the four sibutramine（ 50 mg / kg,100 mg / kg,150 mg / kg and 300 mg / kg） treatment groups,respectively. The general condition of the mice was observed and the survival rate was determined during the drug treatment period. At 15 days after the sibutramine administration,the blood biochemical indicators of cardiotoxicity lactate dehydrogenase（ LDH）,creatine kinase（ CK）,and creatine kinase isoenzyme（ CK-MB） were assessed,and then the mice were sacrificed and heart tissue samples were taken for pathological examination and immunohistochemical observation of the expression of connexin 43（ CX43）. Results（ 1） The blood biochemical indicators in the sibutramine 50 mg / kg and 100 mg / kg Tg（ ＋） treated mice were significantly higher than those in the WT mice（ P〈0. 05 or P〈0. 01）,and significantly higher in the 50 mg / kg,100 mg / kg and 150 mg / kg sibutramine treated Tg（ ＋） mice than in the sT g（ ＋） mice（ P〈0. 05 or P〈0. 01）. However,when the sibutramine was given at a dose of 300 mg / kg,the values of those indicators in the Tg（ ＋） mice were significantly lower than that in the WT and sT g（ ＋） mice（ P〈0. 05 or P〈0. 01）,with the lowest level in the sT g（ ＋） mice.（ 2） The pathological examination revealed cardiotoxic changes in the Tg（ ＋） and WT mice.（ 3） The immunohistochemical analysis showed that alongside with the increasing drug dose,the expression of CX43 was decrease

关 键 词：细胞色素P450 2E1 基因修饰 西布曲明 药物毒性 

分 类 号：Q95-33[生物学—动物学]