目的 研究丹参-红花药对对异丙肾上腺素（isoprenaline hydrochloride，ISO）诱导的心肌缺血大鼠的保护作用及其机制。方法 大鼠随机分为对照组、模型组、复方丹参滴丸（73 mg/kg）组和丹参-红花低、中、高剂量（4、8、16 g/kg）组，给予相应药物干预7 d。于第6天开始ig药物1 h后，大鼠于0℃的冰浴中寒冷刺激4 min，sc ISO（85 mg/kg），连续2 d。采用2,3,5-氯化三苯基四氮唑（2,3,5-triphenyltetrazolium chloride，TTC）染色法测定各组大鼠心肌梗死面积；采用TUNEL法检测各组大鼠心肌细胞凋亡情况；采用苏木素-伊红（HE）染色法观察各组大鼠心肌组织病理变化；采用ELISA法检测各组大鼠血浆中肌酸激酶同工酶（creatine kinase，CK）、天冬氨酸转氨酶（aspartate aminotransferase，AST）、乳酸脱氢酶（lactate dehydrogenase，LDH）、CK-肌红蛋白（myoglobin，MB）活性及心肌肌钙蛋白-I（cardiac troponin-I，CTn-I）、CTn-T、C反应蛋白（C-reaction protein，CRP）、MB、N末端利钠肽前体（N-terminal pro-brain natriuretic peptide，NT-proBNP）和B型脑利钠肽（B-brain natriuretic peptides，BNP）水平；采用Western blotting检测各组大鼠心肌组织中磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase，PI3K）、磷酸化PI3K（phosphorylated PI3K，p-PI3K）、磷酸肌醇依赖性蛋白激酶1（pyruvate dehydrogenase kinase isozyme 1，PDK1）、蛋白激酶B（protein kinase B，Akt）、p-Akt、内皮型一氧化氮合酶（endothelial nitric oxide synthase，eNOS）、p-eNOS、B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）、核因子-κB p65（nuclear factor-κB p65，NF-κB p65）、p-NF-κB p65、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）、细胞色素C（cytochrome C，Cyt C）、半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，Caspase-3）和Caspase-9蛋白表达。结果 丹参-红花药对显著抑制ISO诱导的心肌缺血模型大鼠的心肌梗死面积（P＜0.01）；改善心肌组织病理变化；显著抑制心肌细胞的凋亡（P＜0.05、0.01）；显著降低血浆中心肌酶活性以及心肌蛋白水平（P＜0.05、0.01）；显著上调心肌组织中Bcl-2、PDK1、p-eNOS、p-PI3K和p-Akt蛋白表达水平（P＜0.05、0.01），降低Bax、Cyt C、剪切型Caspase-3（cleaved Caspase-3）、cleaved Caspase-9和p-NF-κB p65蛋白表达水平（P＜0.05、0.01）。结论 丹参-红花药对能够通过调控PI3K/PDK1/Akt信号通路，从而发挥对ISO诱导的心肌缺血的保护作用。

Objectives To study the protective effect and mechanism of Danshen (Salviae Miltiorrhizae Radix et Rhizoma)-Honghua (Carthami Flos) on myocardial ischemia rats induced by isoprenaline hydrochloride (ISO). Methods Rats were randomly divided into control group, model group, Compound Danshen Dropping Pills (73 mg/kg) group and Salviae Miltiorrhizae Radix et Rhizoma-Carthami Flos low-, medium-and high-dose (4, 8, 16 g/kg) groups, rats were given corresponding drug for 7 d. 1 h after ig drug on 6th day, rats were stimulated with cold in ice bath at 0℃ for 4 min and sc ISO (85 mg/kg) for 2 d. 2,3,5-Triphenyltetrazolium chloride (TTC) staining was used to determine the myocardial infarction area of rats in each group; TUNEL method was used to detect myocardial cell apoptosis of rats in each group; Hematoxylin-eosin (HE) staining method was used to observe the pathological changes in myocardial tissue of rats in each group; ELISA method was used to detect creatine kinase (CK), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), CK-myoglobin (MB) activities and cardiac troponin-I (CTn-I), CTn-T, C-reaction protein (CRP), MB, N-terminal pro-brain natriuretic peptide (NT-proBNP), B-brain natriuretic peptides (BNP) levels in plasma of rats in each group; Western blotting was used to detect phosphatidylinositol 3-kinase (PI3K), phosphorylated PI3K (p-PI3K), pyruvate dehydrogenase kinase isozyme 1 (PDK1), protein kinase B (Akt), p-Akt, endothelial nitric oxide synthase (eNOS), p-eNOS, B-cell lymphoma-2 (Bcl-2), nuclear factor-κB p65 (NF-κB p65), p-NF-κB p65, Bcl-2 associated X protein (Bax), cytochrome C (Cyt C), cysteine-aspartate protease-3 (Caspase-3) and Caspase-9 protein expressions in myocardial tissue of rats in each group. Results Salviae Miltiorrhizae Radix et Rhizoma-Carthami Flos significantly inhibited the myocardial infarction area in ISO-induced myocardial ischemia model rats (P < 0.01), improved the pathological changes of myocardial tissue, significantly inhibited the apoptosis of cardiomyocytes (P < 0.05, 0.01), significantly decreased myocardial enzyme activities and myocardial protein levels in plasma (P < 0.05, 0.01), significantly increased protein expression levels of Bcl-2, PDK1, p-eNOS, p-PI3K and p-Akt in myocardial tissue (P < 0.05, 0.01), decreased protein expression levels of Bax, Cyt C, cleaved Caspase-3 (cleaved Caspase-3), cleaved Caspase-9 and p-NF-κB p65 (P < 0.05, 0.01). Conclusion Salviae Miltiorrhizae Radix et Rhizoma-Carthami Flos drug pair can exert a protective effect on ISO-induced myocardial ischemia by regulating PI3K/PDK1/Akt signaling pathway.

R285.5

国家自然科学基金资助项目（81974544）；陕西省科学技术厅资助项目（2019SF-286）；陕西省教育厅资助项目（18JS025）