益气活血方对心肌梗死大鼠冠状动脉内皮细胞损伤的影响

Effect of Yiqi Huoxue recipe on coronary endothelial cell injury in myocardial infarction rats

目的 探讨益气活血方对心肌梗死大鼠冠状动脉内皮细胞损伤的影响。 方法 选取8周龄、健康、雄性无特定病原体级大鼠，采用左冠状动脉前降支结扎或打松结的 方法建立心肌梗死模型和假手术模型（假手术组，20只）；将心肌梗死模型大鼠完全随机分为模型对照组、通心络组、培哚普利组及中药组，每组各23只。假手术组于其他组造模 第1天灌胃给予无菌蒸馏水，模型对照组于造模第1天灌胃给予无菌蒸馏水，通心络组于造模第1天灌胃给予通心络0.25 g/kg，培哚普利组于造模第1天灌胃给予培哚普利0.4 mg/kg ，中药组于造模第1天灌胃给予益气活血方25 g/kg；各组大鼠均采用等容积给药，以无菌蒸馏水调和。采用酶联免疫吸附试验检测5组大鼠造模7、28 d后血清一氧化氮、内皮素1、 前列环素、血栓素A2、可溶性内皮细胞蛋白C受体（EPCR）、可溶性血栓调节蛋白（TM）水平；采用Trizol法提取心肌组织总RNA，通过聚合酶链反应检测EPCR、TM mRNA表达水平， 比较各组间的差异。 结果 造模7、28 d后，模型对照组血清内皮素1、血栓素A2、TM、EPCR水平及心肌组织TM mRNA、EPCR mRNA表达水平均明显高于假手术组［造模7 d： （15.8±1.7）ng/L比（5.3±1.2）ng/L，（27.1±2.5）ng/L比（10.7±0.8）ng/L，（99±8）ng/L比（53±10）ng/L,（351±27）ng/L比（221±21）ng/L，（1.86±0.98）比（ 1.21±0.23），（2.09±0.61）比（1.53±0.11）；造模28 d：（15.6±2.3）ng/L比（6.3±1.7）ng/L，（21.7±7.5）ng/L比（9.6±2.7）ng/L，（110±11）ng/L比（66±8） ng/L，（399±36）ng/L比（205±27）ng/L，（3.71±1.22）比（1.24±0.32），（4.09±1.36）比（1.49±0.29）］（P＜0.01），血清一氧化氮、前列环素水平明显低于假手术 组［造模7 d：（18.6±3.3）ng/L比（30.2±2.6）ng/L，（121±19）ng/L比（187±20）ng/L；造模28 d：（15.2±3.0）ng/L比（31.4±0.0）ng/L，（183±26）ng/L比（220± 19）ng/L］（P＜0.01）；通心络组、培哚普利组及中药组大鼠血清内皮素1、血栓素A2、EPCR、TM水平及心肌组织TM mRNA、EPCR mRNA表达水平均明显低于模型对照组（P＜0.01） ，血清一氧化氮、前列环素水平明显高于模型对照组（P＜0.01）。 结论 益气活血方能减轻心肌梗死大鼠的血管内皮细胞损伤，明显改善内皮细胞功能，从而调节微循环稳态 ，平衡冠状动脉收缩及舒张，最终缓解心肌缺血。

Objective To explore the effect of Yiqi Huoxue recipe on coronary endothelial cell injury in myocardial infarction rats. Methods Left anterior descending coronary artery ligation method or tying loose knot method were used to make rat myocardial infarction model and sham operation model （sham operation group, 20 cases）; myocardial infarction rats were randomly divided into model control group, Tongxinluo group, perindopril group and Yiqi Huoxue recipe group, with 23 rats in each group. The sham operation and model control group were intragastricly administrated sterile distilled water at the first day of molding; the Tongxinluo group was intragastricly administrated Tongxinluo 0.25 g/kg; the perindopril group had perindopril 0.4 mg/kg and the Yiqi Huoxue recipe group had Yiqihuoxue recipe 25 g/kg. Serum levels of nitric oxide（NO）, endothelin-1（ET-1）, prostaglandin I2（PGI2）, thromboxane A2 （TXA2）, soluble endothelial cell protein C receptor（EPCR） and soluble thrombomodulin（TM） 7, 28 d after modeling were determined through enzyme linked immunosorbent assay; EPCR and TM mRNA in myocardial tissue were extracted through Trizol method and EPCR and TM mRNA expression were determined through polymerase chain reaction. Results Serum levels of ET-1, TXA2, TM, EPCR and TM mRNA, EPCR mRNA expressions in myocardial tissue in model control group were significantly higher than those in sham operation group[7 d after molding：（15.8±1.7）ng/L vs （5.3±1.2）ng/L,（27.1±2.5）ng/L vs （10.7±0.8） ng/L,（99±8）ng/L vs （53±10）ng/L,（351±27）ng/L vs （221±21）ng/L,（1.86±0.98） vs （1.21±0.23）,（2.09±0.61） vs （1.53±0.11）;28 d after molding：（15.6±2.3）ng/L vs （6.3±1.7）ng/L,（21.7±7.5）ng/L vs （9.6±2.7）ng/L,（110±11）ng/L vs （66±8）ng/L,（399±36）ng/L vs （205±27）ng/L, （3.71±1.22） vs （1.24±0.32）,（4.09±1.36） vs （1.49±0.29）]（P〈0.01）; serum NO and PGI2 levels in model control group were significantly lower than those in sham operation group [7 d after molding：（18.6±3.3）ng/L vs （30.2±2.6）ng/L,（121±19）ng/L vs （187±20）ng/L;28 d after molding： （15.2±3.0）ng/L vs （31.4±0.0）ng/L,（183±26）ng/L vs （220±19）ng/L]（P〈0.01）. Serum levels of ET-1, TXA2, TM, EPCR and expression levels of TM mRNA, EPCR mRNA in myocardial tissue in Tongxinluo group, perindopril group and Yiqi Huoxue recipe group 7, 28 d after moding were lower and serum NO, PGI2 levels were higher than those in model control group, the differences were significant（P〈0.01）. Conclusion Yiqi Huoxue recipe in myocardial infarction rat model can reduce endothelial cells injury, improve endothelial function, regulate microcirculation homeostasis and relieve myocardial ischemia.