银杏二萜内酯葡胺注射液对离体心脏缺血再灌注损伤保护作用的实验研究

Experimental study on the protective effect of Diterpene Ginkgolides Meglumine injection on isolated heart ischemia-reperfusion injury

关键词:  银杏二萜内酯葡胺注射液  离体心脏  缺血再灌注  血流动力学  ATPase

Key words:  Diterpene Ginkgolides Meglumine injection  isolated heart  ischemia-reperfusion  hemodynamics  ATPase

摘要:

[目的]研究银杏二萜内酯葡胺注射液（DGMI）对大鼠离体心脏缺血再灌注损伤的保护作用并探讨其机制。[方法]取120只SD大鼠随机分为正常对照组、模型组、DGMI低（1.5 mg/kg）、中（3 mg/kg）、高（6 mg/kg）剂量组和维拉帕米2.5 mg/kg组（阳性药物对照组），各20只；手术前2 h，腹腔注射给药，正常对照组和模型组同步给予生理盐水。采用Langendorff灌流系统建立离体心脏灌注模型，正常对照组持续灌注Krebs-Henseleit液（K-H液），模型组、DGMI各剂量组和维拉帕米组平衡灌注20 min后停灌30 min、再灌注60 min后行各指标检测。测定冠脉流出液心肌酶活性；2，3，5-氯化三苯基四氮唑（TTC）染色测定心脏组织梗死面积，苏木精-伊红（HE）染色观察心脏组织病变；测定血流动力学指标；测定心肌组织中抗氧化酶活性以及丙二醛（MDA）含量；测定心肌组织ATPase活力。[结果]与模型组比较，DGMI中、高剂量组和维拉帕米2.5 mg/kg组大鼠血清心肌酶谷草转氨酶（AST）、乳酸脱氢酶（LDH）和肌酸激酶MB同工酶（CK-MB）活性均显著降低（P<0.05或P<0.01）；心脏组织梗死面积显著降低（P<0.01），心脏组织病变明显改善；明显改善血流动力学指标：心率（HR）、左室收缩压（LVSP）、左室收缩压最大上升速率（+dP/dt_max）、左室舒张压最大下降速率（-dP/dt_min）均显著升高且左室舒张末压（LVEDP）显著降低（P<0.05或P<0.01）；抗氧化酶[超氧化物歧化酶（SOD）、过氧化氢酶（CAT）]活性显著升高且MDA含量显著降低（P<0.05或P<0.01）；Na⁺-ATPase、Ca²⁺-ATPase酶活力显著升高（P<0.05或P<0.01）。[结论]DGMI可能通过改善血液流变学、降低氧化应激损伤、改善ATPase活力而对大鼠离体心脏表现出一定的保护作用。

Abstract:

[Objective] To study the protective effect of Diterpene Ginkgolides Meglumine Injection (DGMI) on isolated rat heart ischemia-reperfusion injury and to explore its possible mechanism.[Methods] The 120 SD rats were randomly divided into normal control group, model group and DGMI low-dose (1.5 mg/kg), medium-dose (3 mg/kg), high-dose (6 mg/kg) groups and Verapamil 2.5 mg/kg group (positive control group), 20 in each group. Two hours before surgery, the drugs were given by intraperitoneal injection, normal control and model groups were given saline. The heart perfusion model was established by Langendorff perfusion system, and the normal control group continued infusion of K-H solution, model group and salvia miltiorrhiza polyphenolic acid group were perfused 20 min, and then subjected ischemia for 30 min, followed by a 60 min reperfusion period. The myocardial enzyme activity in coronary effluent were determined; TTC staining was used to measure the myocardial infarct size were measured by TTC staining, the cardiac tissue lesions were observed by HE staining; the hemodynamic parameters were determined; the activities of antioxidant enzymes and malondialdehyde (MDA) in myocardium were measured; the activity of ATPase in myocardium was measured.[Results] Compared with model group, the activity of AST, LDH and CK-MB of DGMI medium, high-dose groups and Verapamil 2.5 mg/kg group were significantly increased (P<0.05 or P<0.01). The cardiac tissue infarct size were significantly reduced (P<0.01), cardiac lesions were significantly improved; the hemodynamic parameters were significant improved:the HR, LVSP, +dP/dtmax,-dP/dtmin were significantly increased and LVEDP was significantly decreased (P<0.05 or P<0.01). The activity of SOD, CAT in myocardium were significantly increased and the content of MDA were significantly decreased (P<0.05 or P<0.01). The activity of Na⁺-ATPase, Ca²⁺-ATPase were significantly increased (P<0.05 or P<0.01).[Conclusion] The DGMI may play a protective role in isolated rat heart by improving hemorheology, reducing oxidative stress injury and improving ATPase activity.