金纳多对心肌保护作用机制的实验研究

Cardioprotective Effects of Ginaton in Ischemia-Reperfusion Rabbits

目的:观察金纳多注射液对兔心肌缺血再灌注后热休克蛋白70的诱导作用及其心肌保护作用的机制。方法:取成功建立心肌缺血30 min再灌注模型的新西兰大白兔30只,根据灌注液的不同随机分为两组:①盐水组(n=15),根据再灌注时间的不同平均(n=5)分为再灌注1 h、3 h和5 h 3种情况;②金纳多组(n=15),根据再灌注时间的不同平均(n=5)分为再灌注1 h、3 h和5 h 3种情况。另取5只为正常对照组(无心肌缺血)。采用western blot蛋白印迹技术检测各组心肌热休克蛋白70的含量,并进行半定量分析;检测各组血清总超氧化物歧化酶、丙二醛及肌酸激酶变化。结果:①心肌组织热休克蛋白70含量:缺血区和非缺血区在正常对照组、盐水组、金纳多组心肌热休克蛋白70均分别显示极少表达、轻度或明显表达及过度表达。缺血区和非缺血区各组间比较均有极显著性差异(P<0．01),各组的缺血区与非缺血区比较(除了盐水组再灌注5 h外)均有显著性差异(P<0．05～0．01)。②血清总超氧化物歧化酶含量:与正常对照组比较,再灌注不同时间的盐水组和金纳多组血清总超氧化物歧化酶含量均降低,且有显著性差异(P<0．05- 0．01);与同一再灌注时间盐水组比较,金纳多组均增高,有极显著性差异(P<0．01)。③血清丙二醛含量:与正常对照组比较,再灌注1 h和3 h时,盐水组和金纳多组血清丙二醛均增高,且有显著性差异(P<0．01或P<0．05),再灌注5 h基本恢复至正常对照组水平。且金纳多组较盐水组明显降低有显著差异(P<0．05或P<0．01)。④血清肌酸激酶含量;与正常对照组比较,盐水组再灌注1 h、3 h、5 h和金纳多组再灌注3 h、5 h血清肌酸激酶含量增高,有极显著性差异(P<0．01);且再灌注不同时间的金纳多组较同一再灌注时间盐水组降低,有显著性差异(P<0．05或P<0．01)。结论:心肌缺血再灌注3小时后热休克蛋白70开始明显表达,金纳多注射液可以诱导热休克蛋白70及早且大量表达,对心肌具有保护作用;金纳多的抗氧化作用可能与热休克蛋白70的表达有关。

Objective： To observe the effects of Ginaton injection on expression of myocardial heat shock protein 70 and the underlying cardioprotective mechanism in ischemia-reperfusion rabbits. Methods： Thirty rabbits with ischemia for 30 minutes received saline or Ginaton reperfusion intravenously for 1h,3h and 5h, respectively. There were five rabbits at every time point. Five rabbits without ischemia-reperfusion were used as normal control group. Western blotting was used to measure heat shock protein 70 semiquantitatively. Serum Total Supcroxide Dismutase,Malondiadehyde, and Creatine Kinase were also measured. The results were analyzed and compared between the two group using t test. Results ： Less expression level of heat shock protein 70 was observed in normal control group. The ratio of ischemic to non-ischemic area was 28. 12 ±0. 73 to 23.42 ±0. 54（ P 〈0. 01 ）. Significantly elevated expression levels of heat shook protein 70 in saline reperfusion group at 3h and Ginaton reperfusion group at lh, the ratio of ischemic to non-ischemic area was 69. 92 ± 1.19 to 49.41 ±0. 7 （ P 〈 0.01 ） and 373.92 ± 4.09 to 275.98± 4.91 （ P 〈 0.05 ）, respectively. The expression level of heat shock protein 70 increased gradually with repeffusion time by folds Serum Total Superoxide Dismutase level obviously descreased and Malondiadehyde level obviously increased in saline group. Serum Total Superoxide Dismutase level slightly decreased and Malon-diadehyde level slightly increased in Ginaton group, there is statistical difference between the two groups at the same time pionts （P 〈 0. 05 or P 〈 0. 01 ）. Serum Creatine Kinase level slightly changed in Ginaton group, but obviously increased in saline group, statistical difference is significant between the two groups at the same points（ P 〈 0. 05 or P 〈 0. 01 ）. Conclusion： Heat shock protein 70 expresses in high level at 3h after ischemia-reperfusion, Ginaton injection can induce early and excessive expression of heat shock protein 70, leading to protect cardiac tissue. Ginaton＇ s anti-oxidation may relate to expression of heat shock protein 70.