Department of Natural Medicinal Chemistry, China Pharmaceutical University, Nanjing, 211198, People's Republic of China.
National Engineering Research Center for Modernization of Traditional Chinese Medicine - Hakka Medical Resources Branch, School of Pharmacy, Gannan Medical University, Ganzhou, 341000, People's Republic of China.
J Ethnopharmacol. 2020 Aug 10;258:112881. doi: 10.1016/j.jep.2020.112881. Epub 2020 Apr 18.
Callicarpa kwangtungensis Chun (C. kwangtungensis) is a very famous herbal medicine with the function of promoting blood circulation and removing blood stasis which is beneficial for cardiovascular disease (CVD). Phenylethanoid glycosides (PGs) are the major class of active ingredients in C. kwangtungensis and present significant anti-oxidative and anti-inflammatory property related to apoptosis. Therefore, this study aimed to investigate the effects of total phenylethanoid glycosides of C. kwangtungensis (CK-PGs) on isoproterenol (ISO) induced myocardial ischemic injury (MI) and the mechanisms related to the apoptosis mediated by oxidative damage and inflammation.
The myocardial ischemia animal model was established as subcutaneous injecting ISO. Echocardiography and biomarkers were employed to determine the degree of myocardial damage. Histopathological changes were observed by hematoxylin and eosin test. The TUNEL staining and activity of caspase-3 were measured to detect the level of apoptosis which is medicated by the oxidative damage detected by the level of MDA, GSH and ROS tested with the kit and the inflammation reflected by TNF-α. The activity of Na-K-ATPase (NKA) was detected by the commercial kits, whose expression was measured by immunohistochemistry analysis. At last, Western blot analysis was used to measure Na-K-ATPase/Src/ERK1/2 and Bax/Bcl-2 pathway.
CK-PGs showed cardioprotective effect against ISO-induced myocardial ischemic injury evidenced by improving heart function and lowering myocardial injury markers. CK-PGs could inhibit the level of apoptosis as shown by the decrease of the TUNEL-positive cells, the activity of caspase-3 and increase of the expression of Bax. CK-PGs also reduced oxidative stress and inflammation to suppress apoptosis by decreasing the level of ROS, MDA, and increasing GSH activity and lowering the level of TNF-α. In addition, CK-PGs exerted the protection by increasing the activity and the expression of NKA. Meanwhile, Na-K-ATPase/Src/ERK1/2pathway was weakened for the inhibition of apoptosis.
CK-PGs could protect cardiomyocytes from myocardial injury through suppressing Na-K-ATPase/Src/ERK1/2 pathway and inhibiting apoptosis mediated by oxidative stress and inflammation.
ethnopharmacological 相关性：黑老虎（C. kwangtungensis）是一种非常著名的草药，具有活血化瘀的作用，有利于心血管疾病（CVD）。苯乙醇苷（PGs）是黑老虎的主要活性成分，具有显著的抗氧化和抗炎特性，与细胞凋亡有关。因此，本研究旨在探讨黑老虎总苯乙醇苷（CK-PGs）对异丙肾上腺素（ISO）诱导的心肌缺血损伤（MI）的影响及其与氧化损伤和炎症介导的细胞凋亡相关的机制。
通过皮下注射 ISO 建立心肌缺血动物模型。采用超声心动图和生物标志物来确定心肌损伤程度。苏木精-伊红（H&E）染色观察组织学变化。通过 TUNEL 染色和 caspase-3 活性检测来检测细胞凋亡水平，细胞凋亡是由氧化损伤导致的 MDA、GSH 和 ROS 水平检测试剂盒以及 TNF-α 反映的炎症介导的。通过商业试剂盒检测 Na-K-ATPase（NKA）的活性，通过免疫组化分析测量其表达。最后，采用 Western blot 分析测定 Na-K-ATPase/Src/ERK1/2 和 Bax/Bcl-2 通路。
CK-PGs 对 ISO 诱导的心肌缺血损伤具有心脏保护作用，表现为改善心功能和降低心肌损伤标志物。CK-PGs 可抑制细胞凋亡水平，表现为 TUNEL 阳性细胞减少、caspase-3 活性降低、Bax 表达增加。CK-PGs 还通过降低 ROS、MDA 水平，增加 GSH 活性，降低 TNF-α 水平，减轻氧化应激和炎症，抑制细胞凋亡。此外，CK-PGs 通过增加 NKA 的活性和表达发挥保护作用。同时，Na-K-ATPase/Src/ERK1/2 通路减弱，抑制细胞凋亡。
CK-PGs 可通过抑制 Na-K-ATPase/Src/ERK1/2 通路，抑制氧化应激和炎症介导的细胞凋亡，保护心肌细胞免受心肌损伤。
