激光共聚焦显微镜观察前胡丙素对新生大鼠缺氧再灌注心肌细胞内游离钙的影响  

The Effect of Praeruptorin C on the Concentration of Free calcium in Cultured Myocardial Cells of Neonatal Rat Exposed to Ischemia and Reperfusion Observed by Laser Confocal Microscopy

作　　者：于晓頔[1] 祝宝华[2] 王金桂[1] 

机构地区：[1]东南大学临床医学院,江苏南京210009 [2]东南大学医学院附属扬州医院心血管病研究所,江苏扬州225001

出　　处：《实用临床医药杂志》2008年第3期38-41,共4页Journal of Clinical Medicine in Practice

基　　金：江苏省135工程医学重点人才课题资助基金项目(RC2002016)

摘　　要：目的激光共聚焦显微镜观察缺氧再灌注损伤对心肌细胞内Ca2+浓度([Ca2+]i)的影响,以及前胡丙素对模拟心肌缺氧再灌注过程中减轻钙超载的作用。方法采用SD大鼠乳鼠进行心肌细胞培养,建立模拟缺氧再灌注模型。实验分4组:正常组:细胞正常培养;缺氧预适应组:预先缺氧2 h,复氧1 h,再缺氧12 h后,复氧2 h;前胡丙素预处理组:先予前胡丙素单体终浓度为100μmol/L作用1 h,再行缺氧12 h,后复氧2 h;模拟缺氧再灌注组缺氧12 h,后复氧2 h。细胞上清检测LDH值。心肌细胞以Fluo-3/AM荧光指示剂负载,应用激光共聚焦显微镜技术检测心肌细胞([Ca2+]i)变化。结果模拟缺氧再灌注组心肌细胞([Ca2+]i)荧光强度值显著高于前胡丙素预处理组及缺氧预适应组(P<0.01),前胡丙素预处理组与缺氧预适应组细胞内荧光强度无明显组间差异。结论心肌细胞缺氧再灌注损伤导致Ca2+超载,而前胡丙素有明显减轻心肌细胞模拟缺氧再灌注时Ca2+超载的作用。应用激光扫描共聚焦显微镜技术可以直观地进行细胞内钙离子研究。Objective To study the effects of praeruptorin C on intracellular calcium concentration in cultured myocardial cells of neonatal SD rat exposed to ischemia and reperfusion, intracellular calcium was detected with Fluo-3/AM by laser confocal microscopy. Cultures of cardiomyocytes were used to establish hypoxia-reoxygenation models by means of reagent chemicals depleting oxygen. Methods The primary cadiomyocytes were randomly divided into 4 groups： the normally cultured group（control）, the hypoxia preconditioning（HP） group, the hypoxia-reoxygenation （HR） group and the praeruptorin C preconditioning （PP） group. Each group was untreated or treated with 100 μmolPL praeruptorin C. The value of LDH of celluar supernatant was detected by automatic biochemistry analyzer. The fluorescence intensity of intracellular Ca^2＋ was detected respectively with Fluo-3/AM loading by the laser confocal microscopy. Result After ischemia reperfusion treatment the intracellular calcium increased remarkably from normal level of 28. 4330 ± 3. 05465 to 43.3290±2. 48112.The concentration of intracellular calcium wasinhibited significantly to 33. 9750 ± 3. 46351 after praeruptorin C treatment. Conclusion The results suggested that hypoxia and reperfusion treatment could induce the calcium overloading in cultured myocardial cells and the praeruptorin C could inhibit that intracellular calcium overloading significantly. Laser confocal microscopy can be used to intuitively observe intracellular Ca^2 ＋ .

关 键 词：心肌缺氧 再灌注 钙 前胡丙素 激光共聚焦显微技术 

分 类 号：R541.4[医药卫生—心血管疾病]