摘要
目的:观察中药黄芩苷对离体状态下缺血再灌注损伤心脏的保护效应。方法:实验于2005-09/12在锦州医学院药理学教研室完成。选用雄性成年SD大鼠60只,头部击昏后迅速取出心脏,采用离体大鼠心脏Lan-gendorff灌流模型制备离体心脏缺血再灌注损伤模型。随机数字表法分为5组,每组12只。①对照组:以含氧K-H液灌注100min。②再灌注组:在对照组基础上灌注含氧K-H液10min后,停止灌流30min,然后再灌注60min。③黄芩苷60μmol/L再灌注组:灌注方法同再灌注组,再灌注时加入60μmol/L黄芩苷溶液。④黄芩苷120μmol/L再灌注组:再灌注时加入120μmol/L黄芩苷溶液。⑤黄芩苷240μmol/L再灌注组:再灌注时加入240μmol/L黄芩苷溶液。应用黄嘌呤测定法测定超氧化物歧化酶活性P应用TAB比色原理测定心肌组织中脂质过氧化产物丙二醛含量P收集各组冠脉流出液测定肌酸激酶、乳酸脱氢酶活性,用以评价心肌细胞受损程度及黄芩苷的保护作用。原位末端标记法标记后应用流式细胞仪进行细胞凋亡率测定,进一步从DNA水平观察细胞受损程度及黄芩苷的保护作用。计量资料差异比较采用t检验,组间数据处理采用方差齐性分析,采用非配对t检验。结果:在实验过程中无动物死亡,全部进入结果分析。①大鼠心肌组织超氧化物歧化酶活性:再灌注组较对照组明显下降[(7301±350),(3400±600)nkat/g,P<0.05];黄芩苷60,120,240μmol/L再灌注组较再灌注组有不同程度增加且随浓度增加作用增强[(5251±717),(5917±172),(6818±633),(3400±600)nkat/g,P<0.05]。②丙二醛含量:再灌注组较正常组增加[(85±17),(275±20)nmol/g,P<0.05],黄芩苷60,120,240μmol/L再灌注组较再灌注组有不同程度下降且呈剂量依赖性[(189±32),(126±27),(97±19),(275±20)nmol/g,P<0.05]。③冠脉流出液中肌酸激酶活性活性变化:再灌注组较正常组增加[(80.28±17.12),(105.43±3.95)mkat/L,P<0.05],黄芩苷60,120,240μmol/L再灌注组较再灌注组降低且有剂量依赖性[(90.86±9.13),(86.90±5.68),(84.40±19.15),(105.43±3.95)mkat/L,P<0.05]。④冠脉流出液中乳酸脱氢酶活性:再灌注组较正常组增加[(47.24±4.56),(68.59±11.74)mkat/L,P<0.05],黄芩苷60,120,240μmol/L再灌注组较再灌注组降低且有剂量依赖性[(50.45±5.37),(49.94±4.86),(47.57±4.58),(68.59±11.74)mkat/L,P<0.05]。⑤大鼠心肌细胞凋亡率:再灌注组较对照组凋亡率明显增加(10.58%,23.73%,P<0.05),黄芩苷60,120,240μmol/L再灌注组比再灌注组降低且呈剂量依赖性(9.35%,15.69%,10.41%,23.73%,P<0.05)。结论:黄芩苷可改善心肌缺血再灌注损伤所致乳酸脱氢酶、肌酸激酶释放量增加及心肌超氧化物歧化酶生成量的降低,降低乳酸脱氢酶、肌酸激酶、丙二醛释放量,增高超氧化物歧化酶活性;减少心肌细胞凋亡率。黄芩苷对离体心脏缺血再灌注所致损伤及凋亡有保护作用,并呈现剂量依赖性。
AIM： To observe the protective effect of baicalin on isolated heart of rats with ischemic-reperfusion injury. METHODS： The experiment was performed in the Laboratory of Pharmacology of Jinzhou Medical College from September to December 2005. Sixty adult male SD rats were selected and knocked down by hitting heads to remove the heart. Langebdorff perfusion model was made in isolated hearts with ischemia-reperfusion injury. Sixty SD rats were randomly divided into five groups with 12 rats in each group.①Control group： Rats were perfused with K-H fluid containing oxygen for 100 minutes. ②Reperfusion group： A 30-minute interval was given after 10-minute perfuion of K-H fluid, and then rats were reperfused for 60 minutes.③60 μmol/L baicalin reperfusion group： rats were perfused the same as reperfusion group with 60 μmol/L of baicalin solution added. ④120 μmol/L baicalin reperfusion group：120 μmol/L baicalin solution was added into when reperfusion. ⑤240 μmol/L baicalin reperfusion group： 240 μmol/L baicalin solution was added into when reperfusion. The activities of plasma superoxide dismutase （SOD） were measured by xanthine oxidase and the contents of serum malondialdehyde （MDA） in myocardial tissues were determined by the method of thiobarbituric acid. The activities of creatine kinase （CK） and lactic dehydrogenase （LDH） in the effluent from arteria coranaria were measured to evaluate the injury of myocardial cells and the protective effects of baicalin. The apoptotic rate was determined by terminal deoxynucleotidyl transferase mediated nick end labeling （Tunel）and flow cytometry （FCM）, so as to further study the cell injury and the protective effects of baicalin from DNA level. The differences of measurement data were compared with t test, and differences among groups were compared with homoscedasticity as well as nonpairing t test. RESULTS： All animals were involved in the analysis of results, and no rat died in the experiment.①The activities of plasma SOD in myocardial tissues of rats： those were significantly decreased in the reperfusion group [（7301±350）, （3400±600） nkat/g, P 〈 0.05], and those in all baicalin reperfusion groups increased with the increased concentration than those in the reperfusion group [（5251±717）,（5917±172）,（6818±633）,（3 40（0600） nkat/g, P 〈 0.05]. ②The contents of serum MDA： Those were increased in the reperfusion group than those in the normal group [（85±17）,（275±20） nmol/g, P 〈 0.05], while those in all baicalin reperfusion groups decreased differently than those in the reperfusion group, which were in dose-dependent manner [（189±32）,（126±27）,（97±19）,（275±20） nmol/g, P 〈 0.05].③The changes in activities of CK in the effluent from arteria coranaria： those were significantly increased in the reperfusion group than those in the normal group [（80.28±17.12）,（105.43±3.95） mkat/L, P 〈 0.05],while those in all baicalin reperfusion groups decreased than those in the reperfusion group, which was in a dose-dependent manner [（90.86±9.13）, （86.90±5.68）, （84.40±19.15）, （105.43±3.95） mkat/L, P 〈 0.05].④The activities of lactic dehydrogenase in the effluent from arteria coranaria： those were remarkably increased in the reperfusion group than those in the normal group [（47.24±4.56）, （68.59±11.74） mkat/L, P 〈 0.05]. and those in all baicalin group groups decreased than those in the reperfusion group, which was in a dose-dependent manner [（50.45 ±5.37）, （49.94 ±4.86）, （47.57 ±4.58）, （68.59±11.74） mkat/L, P 〈 0.05].⑤The apoptotic rate in myocardial cells of rats： it was markedly increased in the reperfusion group than that in the control group （10.58%,23.73%, P 〈 0.05）, while that in all baicalin reperfusion groups were decreased than that in the reperfusion group, which was in a dose-dependent manner （9.35%, 15.69%, 10.41%,23.73% ,P 〈 0.05 ）. CONCLUSION： Baicalin can ameliorate the increased release of LDH and CK induced by ischemia-reperfusion injury and the decrease of SOD in myocardium, decrease the release of LDH, CK and MDA, increase the activities of SOD as well as reduce the apoptotic rate of myocardial cells, which has a significant protective effect on schemia-reperfusion injury and apoptosis of isolated hearts, which are in dose-dependent manner.
出处
《中国临床康复》
CSCD
北大核心
2006年第35期48-50,共3页
Chinese Journal of Clinical Rehabilitation
