水飞蓟宾对离体心脏组织缺血再灌注损伤保护作用及机制研究

Protective effect and mechanism of Silibinin on isolated heart tissue ischemia-reperfusion injury

关键词:  水飞蓟宾  Langendorff灌流  离体心脏  血流动力学  ATP酶

Key words:  Silibinin  Langendorff perfusion  isolated heart  hemodynamics  ATPase

摘要:

[目的]探讨水飞蓟宾（SIL）对Langendorff灌流大鼠离体心脏缺血再灌注损伤的保护作用及其作用机制。[方法]100只SD大鼠随机分为正常对照组、模型组、SIL低[100 mg/（kg·d）]、中[200 mg/（kg·d）]、高[400 mg/（kg·d）]剂量组，各20只；手术前2h灌胃给药，采用Langendorff灌流系统制备大鼠离体心脏灌注模型，模型组与SIL各组灌注K-H液20 min后停灌30 min、然后再灌注60 min后行各指标检测，而正常对照组持续灌注K-H液。测定冠脉流出液心肌酶活性；通过四氮唑（TTC）染色计算心脏组织梗死面积，采用苏木精-伊红（HE）染色对心脏组织进行病理学检查；测定血流动力学指标；测定抗氧化酶活性和丙二醛（MDA）含量；测定心肌组织Na⁺-ATPase、Ca²⁺-ATPase酶活力。[结果]与模型组比较，SIL中、高剂量组心肌酶：谷草转氨酶（AST）、乳酸脱氢酶（LDH）、肌酸激酶MB同工酶（CK-MB）活性显著降低，梗死面积显著降低；病理学检查结果显示：经SIL干预能够明显改善心脏组织病变；血流动力学指标明显改善[心率（HR）、左室收缩压（LVSP）、左室收缩压最大上升速率（+dP/dtmax）、左室舒张压最大下降速率（-dP/dtmin）显著升高，左室舒张压（LVEDP）显著降低]；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；Na⁺-ATPase、Ca²⁺-ATPase酶活力显著升高，上述结果差异具有统计学意义（P<0.05或P<0.01）。[结论]SIL对离体心脏缺血再灌注损伤具有一定的抑制作用，其作用机制可能与SIL能够改善血液流变学指标、抑制氧化应激、提高ATP酶活力有关。

Abstract:

[Objective] To study the protective effect of Silybin (SIL) on Langendorff perfusion rat isolated from ischemia-reperfusion injury and study its mechanism.[Methods] 100 SD rats were randomly divided into normal control group, model group and SIL low-dose, medium-dose, high-dose[100, 200, 400 mg/(kg·d)], n=20. Two hours before surgery, the drugs were given by intragastric administration. The heart perfusion model was established by Langendorff perfusion system, 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, 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 Na⁺-ATPase and Ca²⁺-ATPase in myocardium was measured.[Results] Compared with model group, the activity of AST, LDH and CK-MB of SIL medium-dose, high-dose were significantly increased; the cardiac tissue infarct size were significantly reduced, 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; the activity of SOD, CAT in myocardium were significantly increased and the content of MDA were significantly decreased; the activity of Na⁺-ATPase, Ca²⁺-ATPase were significantly increased; all of the difference above were significant (P<0.05 or P<0.01).[Conclusion] The SIL has a certain inhibitory effect on isolated heart ischemia-reperfusion injury, which perhaps related to its effects of improving hemorheology, reducing oxidative stress injury and improving ATPase activity.