丹参提取物对大鼠心肌缺血后血管新生的保护作用研究

Protective effects of Salvia miltiorrhiza extract on angiogenesis after myocardial ischemia in rats

目的血管新生是治疗缺血性心脏病(ischemic heart disease,IHD)非常有效的一种方法。本研究的主要目的是探究丹参提取物对大鼠心肌缺血后血管新生的保护作用及其机制。方法48只雄性SD大鼠,随机分为假手术组、模型组、丹参提取物组和阳性药组,每组12只。手术结扎左冠状动脉前降支(left anterior descending,LAD)建立心肌缺血模型,造模成功后丹参提取物组以浓度为0.2928 g/kg的丹参提取物药液灌胃,阳性药组以浓度为1 mg/kg瑞舒伐他汀药液灌胃,模型组、假手术组给予生理盐水灌胃。干预28天后通过超声评估心功能,通过激光多普勒评估心脏血流的变化。体外实验采用氧—葡萄糖剥夺法(oxygen-glucose deprivation,OGD)损伤人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs),通过CCK-8实验和Matrigel成管实验评估丹参提取物对HUVECs的保护作用,通过Western blots检测通路蛋白的表达变化。结果丹参提取物不仅可以改善心肌缺血大鼠的心功能(P<0.01),增加心脏血流量(P<0.01),改善心肌细胞形态和炎症,还可以增加内皮细胞的活力(P<0.05),促进细胞体外成管。机制上丹参提取物能够促进OGD诱导的内皮细胞分泌骨形态发生蛋白2(bone morphogenetic protein-2,BMP2)(P<0.01),下调Delta-Like配体蛋白-4(Delta-like 4,Dll4)和Notch1蛋白的表达(P<0.01,P<0.01)。结论丹参提取物可能通过调节BMP2-Dll4/Notch1通路促进血管新生进而保护心功能。

Objective Angiogenesis is an effective way to treat ischemic heart disease(IHD).The objective of this study was to investigate the protective effect and its mechanism of Salvia miltiorrhiza extract on angiogenesis after myocardial ischemia in rats.Methods Forty-eight male SD rats were randomly divided into sham group,model group,salvia miltiorrhiza extract group and positive drug group,with 12 rats in each group.The myocardial ischemia model was established by surgical ligation of the left anterior descending artery(LAD).After the model was successfully made,the salvia miltiorrhiza extract group was treated with a concentration of 0.2928 g/kg Salvia miltiorrhiza extract and the positive treatment group was treated with a concentration of 1 mg/kg rosuvastatin.Rats in the model group and sham group were given normal saline intragastric administration.After 28 days of intervention,the cardiac function was evaluated by ultrasound,and the changes of cardiac blood flow were evaluated by laser doppler.In vitro experiments,oxygen-glucose deprivation(OGD)was used to damage human umbilical vein endothelial cells(HUVECs).The protective effects of Salvia miltiorrhiza extracts on HUVECs were evaluated by CCK-8 assay and matrigel tube formation assay,and the changes of pathway protein expressions were detected by Western blots.Results Salvia miltiorrhiza extract could not only improve cardiac function(P<0.01),increase cardiac blood flow in rats(P<0.01)and improve cardiomyocyte morphology and inflammation with myocardial ischemia,but also increase the vitality of endothelial cells(P<0.05)and promote cell tube formation in vitro.Salvia miltiorrhiza extract could promote the secretion of BMP2 protein by OGD-induced endothelial cells(P<0.01)and down-regulate the expressions of Dll4 and Notch1 protein(P<0.01,P<0.01).Conclusion Salvia miltiorrhiza extract could promote angiogenesis and protect cardiac function by regulating BMP2-Dll4/Notch1 pathway.