目的 探讨芪参胶囊(Qishen capsule,QSC)对陈旧性心肌梗死大鼠的心肌保护作用及相关的作用机制。方法 前降支结扎法制备大鼠心肌梗死模型,分为以下3组:①陈旧性心肌梗死模型组(MI)(n=14),生理盐水灌胃;②低剂量对照组(MI+QSCL)(n=14),低剂量芪参胶囊混悬液灌胃治疗,5×104 mg·kg-1·d-1,持续4周;③治疗组(MI+QSC)(n=14),常规剂量芪参胶囊混悬液灌胃治疗,10×104 mg·kg-1·d-1,持续4周。Sham组,单纯开胸,冠状动脉左前降支出穿线不行结扎(n=16),其中又分为:①Sham组(n=8):生理盐水灌胃;②Sham+QSC组(n=8):芪参胶囊混悬液灌胃治疗,10×104 mg·kg-1·d-1,持续4周。超声心动评价大鼠心功能,病理学方法计数毛细血管及小动脉的密度并用免疫印迹法(Western blot)评价心肌细胞凋亡和血管新生情况。结果 常规剂量芪参胶囊治疗后可有效改善心功能。与模型组相比, 其左心室射血分数(LVEF)与左室短轴缩短率(LVFS)显著升高,左心室舒张末期内径(LVEDD)与左心室收缩末期内径(LVEDS)明显降低(P<0.05); MI+QSC组的LVEDP与dp/dtmax明显增高。常规剂量芪参胶囊明显抑制心肌细胞的凋亡。与模型组和低剂量组(5×104 mg·kg-1·d-1)相比,常规剂量(10×104 mg·kg-1·d-1)芪参胶囊组梗死面积及梗死瘢痕明显减少,活性caspase-3、Bax、cytochrome C的表达量明显降低,而Bcl-2的表达量明显增加,且Bcl-2/Bax的比值也明显增加。③QSC的干预治疗能够促进心肌梗死后大鼠的血管新生。常规剂量(10×104 mg·kg-1·d-1)芪参胶囊组增加了梗死区平滑肌激动蛋白和CD31阳性的血管密度,明显提高了血管新生水平。分别使用ELISA与实时定量PCR的方法检测了血管内皮生长因子(VEGF)蛋白水平与mRNA水平的表达情况,发现常规剂量芪参胶囊组(10×104 mg·kg-1·d-1)较模型组和低剂量组(5×104 mg·kg-1·d-1)VEGF的表达量显著增加;实时定量PCR检测结果显示,经QSC治疗后,缺氧诱导因子(HIF-1α)的mRNA水平也明显高于模型组和低剂量对照组。VEGF和缺氧诱导因子(HIF-1α)mRNA水平的表达量也明显升高。AKT与MEK/ERK通路相关蛋白的表达量检测结果显示,常规剂量(10×104 mg·kg-1·d-1)芪参胶囊的干预治疗同样增加了磷酸化AKT的表达水平并下调了磷酸化的MEK/ERK的表达水平。结论 低剂量芪参胶囊干预的结果与模型组差异不显著,而常规剂量的芪参胶囊治疗则能够有效改善心肌梗死大鼠的心功能,其潜在的机制为促进梗死周围血管新生,降低心肌细胞的凋亡,提高心肌的抗凋亡能力。这种作用可能是通过激活AKT通路并抑制磷酸化MEK/ERK的表达水平而发挥的。
Abstract
OBJECTIVE To explore whether QSC can have cardioprotective efficacy after myocardial infarction(MI),we design this experiment in a preclinical model of myocardial infarction in rats. METHODS Four weeks after left anterior descending coronary artery ligation, rats received either intragastric administration of QSC,or the same volume of saline.Male SD rats were given the ligation of anterior descending coronary artery. After 4 weeks,42 male rats were chosen and randomly divided into 3 groups. The model group(normal saline solution for 4w, n=14), low-dose control group(5×104 mg·kg-1·d-1 of drug QSC, for 4w, n=14), and QSC moderate dose group(10×104 mg·kg-1·d-1 of QSC for 4w,n=14). The sham-operation group(n=16) was only given treatment of chest-open without ligation of anterior descending coronary artery,divided into 2 groups,the Sham-group(normal saline solution for 4w,n=8)and Sham+QSC group(5×104 mg·kg-1·d-1 of drug QSC,for 4w,n=8).Cardiac function was assessed echo cardio-graphically.Angiogenesis and apoptosis were detected using histology and Western blot methods four weeks after QSC therapy. RESULTS Reductions in infarction area and scar collagen content in MI+QSC group were observed in the infarct region compared with the saline control and MI+QSCL group. QSC also improved cardiac function after treatment. QSC significantly decreased apoptosis relative to control group, evidenced by influencing the expression of Bcl-2, Bax, cytochrom C and caspase-3 in the myocardial infarction. Meanwhile, angiogenesis in the infarctive regions were significantly enhanced relative to control group, evidenced by increased density of a-smooth muscle actin and CD31 positive vessels respectively. Similarly, the mRNA expressions of VEGF and HIF-1α were up-regulated.Additionally. OSC involvement also increased the phosphorylation of AKT and down-regulated the phosphorylation of MEK/ERK. CONCLUSION There is no significant difference between QSC low-dose group and the model group, but QSC moderate dose group can improve cardiac function in rats after MI, the underlying mechanism of which can be explained by increasing angiogenesis,reducing apoptosis partially via the activation of AKT signaling pathway and inhibition of phosphorylation of MEK/ERK.
关键词
芪参胶囊 /
血管新生 /
细胞凋亡
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Key words
Qishen capsule /
angiogenesis /
apoptosis
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参考文献
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脚注
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基金
国家自然科学基金资助项目(81071177)
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