川芎嗪对阿霉素所致体外心肌细胞损伤的保护作用

Protective effect of ligustrazin on cardiomyocyte injury induced by doxorubicin in vitro

目的:心脏毒性限制了阿霉素在肿瘤化疗中的应用,体外实验观察川芎嗪对阿霉素所致心肌细胞损伤的保护作用及其机制。方法:实验于2006-10/2007-02在辽宁医学院药理学教研室完成。①实验材料及分组:选用出生1～3d的SD大鼠30只,雌雄不拘。采用纯化培养的心肌细胞建立阿霉素损伤模型,培养4d的心肌细胞随机分为5组:正常对照组(正常细胞培养,加入等数量的培养基,不加入任何药物),阿霉素损伤组(加入阿霉素1mg/L),阿霉素+川芎嗪低浓度组(加入阿霉素1mg/L和川芎嗪5mg/L),阿霉素+川芎嗪中浓度组(加入阿霉素1mg/L和川芎嗪10mg/L),阿霉素+川芎嗪高浓度组(加入阿霉素1mg/L和川芎嗪20mg/L)。作用48h后检测相关指标。②实验评估:采用乳酸脱氢酶试剂盒测其释放量;采用黄嘌呤氧化酶法测定超氧化物歧化酶活力;硫代巴比妥酸显色法测定丙二醛含量;硝酸还原法测定一氧化氮含量;四唑盐比色法测定心肌细胞线粒体酶活性。③组间显著性检验采用单因素方差分析和LSD检验。结果:①丙二醛含量:阿霉素损伤组明显高于正常对照组(P<0.01),阿霉素+高、中、低浓度川芎嗪组明显低于阿霉素损伤组(P<0.01)。②心肌细胞超氧化物歧化酶活性:阿霉素损伤组明显低于正常对照组(P<0.01),阿霉素+高、中、低浓度川芎嗪组明显高于阿霉素损伤组(P<0.01)。③心肌细胞线粒体酶活性:阿霉素损伤组心肌细胞线粒体酶活性明显低于正常对照组,阿霉素+高、中、低浓度川芎嗪组明显高于阿霉素损伤组(P<0.01)。④心肌细胞内一氧化氮含量:阿霉素损伤组明显高于正常对照组(P<0.01),阿霉素+高、中、低浓度川芎嗪组明显低于阿霉素损伤组(P<0.01)。结论:川芎嗪具有对阿霉素所致心肌细胞氧化损伤的保护作用,可能与其抗氧化、抗自由基作用有关。

AIM： Cardiotoxicity inhibits the application of doxorubicin in oncosis chemotherapy. The present study was designed to investigate the protective effect and mechanism of ligustrazin on cardiomyocyte injury induced by doxorubicin. METHODS： The experiment was performed in the Department of Pharmacology, Liaoning Medical University from October 2006 to February 2007. （1）Totally 30 SD rats aged 1-3 days of either sex were used for the experiment. Doxorubicin injured model was established with purified cultured cardiomyocytes, and the experiment was divided into 5 groups： normal control group （normal cells were cultivated with only medium of the same volume）, doxorubicin injured group （1 mg/L doxorubicin）, doxorubicin plus low concentration ligustrazin group （5 mg/L ligustrazin ＋ 1 mg/L doxorubicin）, doxorubicin plus middle concentration ligustrazin group （10 mg/L ligustrazin ＋ 1 mg/L doxorubicin）, doxorubicin plus high concentration ligustrazin group （20 mg/L ligustrazin ＋ 1 mg/L doxorubicin）. Relevant indexes were measured 48 hours later. （2）The releasing amount, activity of superoxide dismutase （SOD）, contents of malondialdehyde （MDA） and nitrogen monoxide and activity of dehydrogenase in mitochondria were measured by lactic dehydrogenase （LDH）, xanthine oxidase, thiobarbituric acide, nitric acid disoxidation and tetrazolium methods, respectively. （3）The single factor analysis of variance and LSD test were used for statistical analysis. RESULTS： （1）Content of MDA was significantly higher in the doxorubicin injured group than the normal control group, and doxorubicin plus low, middle, high concentration ligustrazin groups （P 〈 0.01）. （2）Activity of SOD was significantly lower in the doxorubicin injured group than the normal control group, and doxorubicin plus low, middle, high concentration ligustrazin groups （P 〈 0.01 ）. （3）Activity of dehydrogenase in mitochondria was significantly lower in the doxorubicin group than the normal control group, and doxorubicin plus low, middle, high concentration groups （P 〈 0,01）. （4）Content of nitrogen monoxide in cardiomyocytes was obviously higher in the doxorubicin injured group than the normal control group, and doxorubicin plus high, middle, low concentration ligustrazin groups （P 〈 0.01）. CONCLUSION： Ligustrazin has a protective effect on cardiomyocytes injured by doxorubicin, which may be associated with its antioxidation and anti-free radical function.