丹酚酸B诱导血管新生对大鼠心肌缺血再灌注损伤的保护机制及凋亡相关蛋白研究

Protective Mechanism and Apoptosis Related Protein of Salvianolic Acid B Induced Angiogenesis on Myocardial Ischemia-Reperfusion Injury in Rats

目的:探讨丹酚酸B诱导血管新生对大鼠心肌缺血再灌注损伤(MIRI)的保护机制及凋亡相关蛋白的影响。方法:40只SD大鼠随机分为假手术组、模型组、培哚普利组和丹酚酸B组,每组10只。假手术组大鼠仅开胸分离左冠状动脉前支,其余大鼠均进行心肌缺血再灌注损伤(MIRI)造模,开胸分离左冠状动脉前降支后,血流阻断0.5 h,再灌注处理2 h。培哚普利组大鼠在造模前3 d腹腔注射培哚普利片溶液,0.4 mg/kg,1次/d;丹酚酸B组大鼠在造模前3 d腹腔注射丹酚酸B溶液,50 mg/kg,1次/d;模型组和假手术组大鼠给予等量生理盐水。各组大鼠灌注2 h后即刻取材测定心肌梗死面积,TUNEL法测定心肌细胞凋亡率,酶联免疫吸附法检测大鼠血清VEGF、TAX2、PGI2水平,Western blotting法测定心肌组织中Caspase-3、PI3K、Bcl-2、Bax蛋白表达水平。结果:模型组大鼠心肌组织总面积、心肌组织梗死面积、梗死面积比例和心肌细胞凋亡率均高于假手术组(P<0.05);培哚普利组和丹酚酸B组大鼠心肌组织梗死面积、梗死面积比例和心肌细胞凋亡率均低于模型组(P<0.05);丹酚酸B组大鼠心肌组织总面积、心肌组织梗死面积、梗死面积比例和心肌细胞凋亡率与培哚普利组比较,差异均无统计学意义(P>0.05)。模型组大鼠血清中VEGF、PGI2含量低于假手术组(P<0.05),TAX2含量高于假手术组(P<0.05);培哚普利组和丹酚酸B组大鼠血清中VEGF、PGI2含量高于模型组(P<0.05),TAX2含量低于模型组(P<0.05);丹酚酸B组大鼠血清中VEGF含量高于培哚普利组(P<0.05),TAX2含量低于培哚普利组(P<0.05),而PGI2含量与培哚普利组比较,差异无统计学意义(P>0.05)。模型组大鼠心肌组织中Caspase-3、PI3K、Bcl-2、Bax蛋白相对表达量均高于假手术组(P<0.05);培哚普利组和丹酚酸B组大鼠心肌组织中Bcl-2蛋白相对表达量高于模型组,Caspase-3、PI3K、Bax蛋白相对表达量低于模型组(P<0.05);丹酚酸B组大鼠心肌组织中Caspase-3、PI3K、Bcl-2、Bax蛋白相对表达量与培哚普利组比较,差异均无统计学意义(P>0.05)。结论:丹酚酸B对心肌缺血再灌注损伤的保护作用可能通过诱导血管新生、抗凋亡实现。

Objective:To investigate the protective mechanism of salvianolic acid B-induced angiogenesis on myocardial ischemia-reperfusion injury(MIRI)in rats and the effect of apoptosis-related proteins.Methods:A total of 40 SD rats were randomly divided into sham operation group,model group,perindopril group and salvianolic acid B group,with 10 rats in each group.In the sham operation group,only the anterior branch of the left coronary artery was separated by thoracotomy.The rest of the rats were subjected to myocardial ischemia reperfusion injury(MIRI)model.Blood flow was blocked for 0.5 h after thoracotomy and reperfusion for 2 h.Three days before modeling,rats in the perindopril group were intraperitoneally injected with perindopril tablet solution,0.4 mg/kg,once a day;rats in the salvianolic acid group B were intraperitoneally injected with salvianolic acid B solution,50 mg/kg,once a day;rats in the model group and sham operation group were given the same amount of normal saline at the same time.The myocardial infarct size was measured immediately after 2 hours of reperfusion.The apoptosis rate of cardiomyocytes was measured by TUNEL.The serum levels of VEGF,TAX2 and PGI2 were measured by ELISA.The protein expression levels of Caspase-3,PI3 K,Bcl-2 and Bax were measured by Western blotting.Results:The total area of myocardial tissue,myocardial tissue infarct area,infarct area ratio,and myocardial cell apoptosis rate in the model group were higher than those in the sham operation group(P<0.05).The myocardial tissue infarct area,infarct area ratio and myocardial cell apoptosis rate in the perindopril group and salvianolic acid B group were lower than those in the model group(P<0.05).There were no significant differences in the total area of myocardial tissue,infarct area,infarct area ratio and apoptosis rate between salvianolic acid B group and perindopril group(P>0.05).The serum levels of VEGF and PGI2 in the model group were lower than those in the sham operation group(P<0.05),and the TAX2 content was higher than that in the sham operation group(P<0.05).The levels of VEGF and PGI2 in the serum of rats in the perindopril group and salvianolic acid B group were higher than those in the model group(P<0.05),and the content of TAX2 was lower than that in the model group(P<0.05).The content of VEGF in salvianolic acid B group was higher than that in perindopril group(P<0.05),the content of TAX2 in salvianolic acid B group was lower than that in perindopril group(P<0.05),but the content of PGI2 in salvianolic acid B group was not significantly different from that in perindopril group(P>0.05).The relative expressions of Caspase-3,PI3 K,Bcl-2,and Bax p rotein in the myocardial tissue of the model group were higher than those in the sham operation group(P<0.05).The relative expression of Bcl-2 protein in myocardial tissue of rats in the perindopril group and salvianolic acid B group was higher than that in the model group,and the relative expression of Caspase-3,PI3 K,and Bax protein was lower than that in the model group(P<0.05).There was no significant difference in the relative expression of Caspase-3,PI3 K,Bcl-2 and Bax proteins between salvianolic acid B group and perindopril group(P>0.05).Conclusion:The protective effect of salvianolic acid B on myocardial ischemia-reperfusion injury may be achieved by inducing angiogenesis and anti-apoptosis.