川芎嗪预处理对心肌细胞缺血/再灌注损伤延迟保护作用

Delayed effects of TMPZ on neonatal rat myocardiocytes subjected to anoxia/reoxygenation

目的探讨川芎嗪预处理对心肌缺血／再灌注损伤延迟保护作用。方法实验用SD新生（1～3d）大鼠，雌雄不拘，无菌取出乳鼠心脏，经分离附着组织、胰蛋白酶消化及纯化制成心肌细胞培养，第4天随机分为4组：正常对照（Control）组、缺氧／复氧（A／R）组、缺氧预处理（APC）组、川芎嗪（TMPZ）组。观察心肌细胞搏动频率、细胞存活率（MTT法）、培养液中乳酸脱氢酶（LDH）活性。结果①细胞搏动频率：缺影复氧组（13±3）次／min，与正常对照组相比差异有显著性（P〈0．05）；APC组（84±6）次／min，TMPZ组（85±3）次／min，对正常心肌细胞搏动频率无明显影响（P〉0．05），与缺氧／复氧组比较差异有显著性（P〈0．05）；APC组、TMPZ组之间差异无显著性（P〉0．05）；②细胞存活率：缺氧／复氧组（36．23±4．05）％，与正常对照组比较，差异有显著性（P〈0．05）；APC组（83．76±6．92）％，TMPZ组（80．56±4．65）％分别与缺氧／复氧组比较，差异有显著性（P〈0．05），APC组、TMPZ组之间差异无显著性（P〉0．05）；③LDH活性：缺氧／复氧组（36．73±2．35）U／L与正常对照组比较，差异有显著性（P〈0．05）；APC组（5．70±0．63）U／L、TMPZ组（5．74±0．34）U／L，分别与缺氧／复氧组比较，差异有显著性（P〈0．05），APC组、TMPZ组之间差异无显著性（P〉0．05）。结论从细胞水平证实TMPZ预处理后24h心肌细胞对再次缺氧／复氧损伤具有保护作用。

Objective To investigate the delayed myocardioprotection of TMPZ pre - conditioning. Methods Cultured myocardial cells of neonatal SD rats have been randomly divided into 4 groups： Control group; anoxia reoxygenation （A/R）group;anoxia preconditioning group （APC） ; TMPZ group; In each group, the cardiomyocytes were subjected to anoxia （3 hr） followed by reoxygenation （1 hr） 24 hr after anoxic （ or TMPZ） preconditioning on the model of the cultured cardiomyocytes. At the end of experiment, myocardial cells beating rate, cell viability and the activity of LDH in culture were measured. Results ①Beating rate of ventricular myocytes： The beating rate was significantly slow in A/R group（ 13 ± 3 beats/min） compared with that of control group （ P 〈 0.05 ） ; Between APC group （ 84 ± 6 beats/min ） and TMPZ group （ 85 ± 3 ） beats/min, the beating rate was similar to each other（ P 〉 0.05 ） , and compared with A/R , the beating rate was significantly increased（ P 〈 0.05 ） ; ②the cell viability ： The cell viability of A/R （ 36.23 ± 4.05 ） % was significantly decreased when compared with that of control group （ P 〈 0.05 ） ; the viability of APC （ 83.76 ± 6.92 ） % , TMPZ （ 80.56 ± 4.65 ） % was significantly increased when compared with A/R respectively, （ P 〈 0.05 ）, between APC and TMPZ, the cell viability was similar to each other（ P 〉 0.05 ） ; ③LDH activity ： LDH activity of A/R （ 36.73±2.35 ） U/L was significantly increased when compared with Control group （ P 〈 0.05 ） ; LDH activity of APC （5.70 ± 0.63 ） U/L or TMPZ （5.74 ± 0.34 ） U/L was significantly decreased when compared with A/R respectively （ P 〈 0.05 ）, but LDH activity of TMPZ was similar to that of APC group（ P 〉 0.05 ）. Conclusion TMPZ preconditioning can mimic the delayed protection on cultured neonatal rat myocardiocytes with hypoxia/repeffusion injury.