Huang Wendong, Yang Yongfei, Zeng Zhi, Su Meiling, Gao Qi, Zhu Banghao
Department of Pharmacology, Cardiac and Cerebral Vascular Research Center, Zhongshan School of Medicine, Sun Yat‑sen University, Guangzhou, Guangdong 510080, P.R. China.
Mol Med Rep. 2016 Nov;14(5):4537-4544. doi: 10.3892/mmr.2016.5822. Epub 2016 Oct 11.
Salvia miltiorrhiza and ligustrazine are traditional Chinese medicines that have been used in combination for treatment of cardiovascular disease, including coronary heart disease, cardiac angina and atherosclerosis in Asia, in particular, China. The present study aimed to determine the effect of S. miltiorrhiza and ligustrazine injection (SLI) on myocardial ischemia/reperfusion (I/R) and hypoxia/reoxygenation (H/R) injuries via the Akt serine/threonine kinase (Akt)‑endothelial nitric oxide synthase (eNOS) signaling pathway. Male Sprague‑Dawley rats were randomly assigned into six groups: i) Sham group; ii) I/R group; iii) Low‑SLI group (6.8 mg/kg/day, i.p.); iv) Medium‑SLI group (20.4 mg/kg/day, i.p.); v) High‑SLI group (61.2 mg/kg/day, i.p.); vi) verapamil group (6 mg/kg/day, i.p.). Prior to surgery, the aforementioned groups were pretreated with a homologous drug once per day for 3 days. The effect of SLI following 35 min coronary artery occlusion and 2 h reperfusion was evaluated by determining infarct size, hemodynamics, biochemical values and histological observations. Additionally, cell viability, caspase‑3 expression, B cell leukemia/lymphoma‑2 (Bcl‑2)/Bcl‑2‑associated X protein (Bax) ratio, phosphorylated (p‑)Akt and p‑eNOS were also investigated following 2 h simulated ischemia and 2 h simulated reperfusion in H9C2 cardiomyocyte cells. Pretreatment with SLI significantly improved cardiac function in a dose‑dependent manner and reduced myocardial infarct size, creatine kinase, lactate dehydrogenase and malondialdehyde levels in blood serum. Additionally, myocardial histopathology changes in the rat model were also alleviated in SLI treatment groups. The present in vitro study revealed that treatment with SLI reduced the apoptotic rate of H9C2 cells by inhibiting the activation of caspase‑3 and increasing the Bcl‑2/Bax ratio. The effect of SLI was associated with increased phosphorylation of the survival kinase Akt at Ser473 and its downstream target eNOS following H/R. The present study determined that SLI may alleviate I/R injury in cardiomyocytes and inhibit apoptosis in rats by the activation of the Akt‑eNOS signaling pathway, and downregulation of the expression levels of proapoptotic factors, including caspase-3.
丹参和川芎嗪是传统中药，在亚洲尤其是中国，它们常联合用于治疗心血管疾病，包括冠心病、心绞痛和动脉粥样硬化。本研究旨在通过Akt丝氨酸/苏氨酸激酶（Akt）-内皮型一氧化氮合酶（eNOS）信号通路，确定丹参川芎嗪注射液（SLI）对心肌缺血/再灌注（I/R）和缺氧/复氧（H/R）损伤的影响。将雄性Sprague-Dawley大鼠随机分为六组：i）假手术组；ii）I/R组；iii）低剂量SLI组（6.8毫克/千克/天，腹腔注射）；iv）中剂量SLI组（20.4毫克/千克/天，腹腔注射）；v）高剂量SLI组（61.2毫克/千克/天，腹腔注射）；vi）维拉帕米组（6毫克/千克/天,腹腔注射）。手术前，上述各组每天用同源药物预处理一次，共3天。通过测定梗死面积、血流动力学、生化指标和组织学观察，评估35分钟冠状动脉闭塞和2小时再灌注后SLI的作用。此外，在H9C2心肌细胞中进行2小时模拟缺血和2小时模拟再灌注后，还研究了细胞活力、半胱天冬酶-3表达、B细胞淋巴瘤-2（Bcl-2）/Bcl-2相关X蛋白（Bax）比值、磷酸化（p-）Akt和p-eNOS。SLI预处理以剂量依赖的方式显著改善心脏功能，并降低血清中心肌梗死面积、肌酸激酶、乳酸脱氢酶和丙二醛水平。此外，SLI治疗组大鼠模型的心肌组织病理学变化也得到缓解。目前的体外研究表明，SLI治疗通过抑制半胱天冬酶-3的激活和增加Bcl-2/Bax比值，降低了H9C2细胞的凋亡率。SLI的作用与H/R后存活激酶Akt在Ser473位点的磷酸化增加及其下游靶点eNOS有关。本研究确定，SLI可能通过激活Akt-eNOS信号通路，下调包括半胱天冬酶-3在内的促凋亡因子的表达水平，减轻心肌细胞的I/R损伤并抑制大鼠细胞凋亡。
