蒙古族药朱日很滴丸对缺血再灌注损伤及凝血功能的影响

目的:探讨朱日很滴丸对缺血/再灌流损伤及凝血功能的影响。方法:体外培养H9c2(2-1)大鼠心肌细胞,Na2S2O4诱导缺氧/复氧损伤,观察朱日很滴丸含药血清对H9c2细胞细胞活力、丙二醛(malondialdehyde,MDA)含量、超氧化物歧化酶(superoxide dismutase,SOD)活性的影响。Wistar大鼠随机分为6组,分别是正常组、模型组、阳性给药组(复方丹参滴丸,72.9 mg·kg~(-1))、朱日很滴丸高、中、低剂量组(248,744,1 240 mg·kg~(-1))。采用Langendorff法灌流离体大鼠心脏,除空白组外,停灌30 min后再灌30 min,造成心肌缺血再灌注损伤模型。于大鼠左心房插入气囊导管,记录朱日很滴丸对离体心脏血流动力学指标的影响,测定不同时间点冠脉流出液肌酸激酶(creatine kinase,CK),乳酸脱氢酶(lactate dehydrogenase,LDH)的活性。健康家兔40只,分为正常组,复方丹参滴丸组(丹参滴丸37.8 mg·kg~(-1)),朱日很滴丸组低、中、高剂量组(129,387,645 mg·kg~(-1)),每组8只。连续给药10 d后,四通道血凝仪测定凝血酶原时间(prothrombin time,PT),活化部分凝血活酶时间(activated partial thromboplastin time,APTT),凝血酶时间(thrombin time,TT)和纤维蛋白原(fibrinogen,FIB)。结果:与模型组比较,朱日很滴丸含药血清组能明显增加缺氧/复氧损伤的H9c2细胞的活力(P0.01,P0.05),升高SOD活力,降低MDA含量(P0.05)。心肌缺血再灌注30 min后,与正常组比较,模型组大鼠左心室收缩压(left ventricular systolic,LVSP),左心室内压变化速率(±dp/dt_(max))明显下降(P0.01,P0.05),左心室舒张末期压力(LVDP)明显增加(P0.01),灌流液中LDH,CK的含量明显增加(P0.01);朱日很高、中、低剂量组LVSP,±dp/dt_(max)明显增加,LVDP明显降低,灌流液中LDH,CK的含量明显降低(P0.01,P0.05)。朱日很高、中、低剂量家兔组FIB明显降低,TT明显延长(P0.01,P0.05)。结论:朱日很滴丸能够改善心肌舒缩功能,保护心肌组织,对心肌缺血再灌注损伤具有保护作用;能够延长凝血时间,有抗凝作用。

Effects of Mongolian Zhurihen drop pills on Ischemia/Reperfusion Injury and Coagulation Function

Objective: To investigate the effect of the Mongolian Zhurihen drop pills on ischemia/reperfusion injury and coagulation function. Method: The Na₂S₂O₄-based hypoxia/reoxygenation injury models were established by H9c2 cells in vitro. The effects of Zhurihen drop pills on H9c2 cells viability, superoxide dismutase (SOD) activity, and malondialdehyde (MDA) activity were observed. Wistar rats were randomly divided into 6 groups:normal group;model group;positive drug group (Danshen drop pill group 72.9 mg·kg^-1); Zhurihen drop pills low, middle, high dose groups (248, 744, 1 240 mg·kg^-1). The hearts of rats were isolated and perfused with Langendorff apparatus. Except the rats of normal group, all the other rats were reperfused for 30 min after 30 min of global ischemia, resulting in the myocardial ischemia-reperfusion injury (I/R) models. Balloon catheter was inserted into the left atrium, then myocardial hemodynamic parameters were monitored and recorded, and the biochemical parameters such as lactate dehydrogenase (LDH) and creatine kinase (CK) in perfusate were measured as well. 40 healthy rabbits were divided into normal group, positive control group (Danshen pill group 37.8 mg·kg^-1);Zhurihen drop pills low, middle, high dose groups (129, 387, 645 mg·kg^-1) (n=8 in each group). After administration for 10 consecutive days, prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and fibrinogen (FIB) were measured with four channel coagulation analyzer. Result: Compared with the model group, H9c2 cells viability was significantly increased (P < 0.01, P < 0.05), SOD activity improved, and MDA content was decreased (P < 0.05) by the Zhurihen drop pills medicated serum. After I/R for 30 min, compared with the normal group, the rats in model group showed significant decline in left ventricular systolic pressure (LVSP) and positive and negative maximal values of the first derivative of left ventricular pressure (±dp/dt_max)(P < 0.01, P < 0.05), significant increase in left ventricular end diastolic pressure (LVDP)(P < 0.01), significant increase in levels of LDH and CK in perfusate(P < 0.01). Zhurihen drop pills low, middle, and high dose groups could significantly increase LVSP and ±dp/dt_max levels, significantly reduce LVDP level, LDH and CK content in perfusate (P < 0.01, P < 0.05). Conclusion: Zhurihen drop pills can protect cardiomyocyte against hypoxia/reoxygenation injury, improve heart function impairment, protect cardiac tissues, and prolong the coagulation time with anticoagulant effects.