Chen G, Luo S
Department of Cardiology, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Feb 20;44(2):226-235. doi: 10.12122/j.issn.1673-4254.2024.02.04.
To investigate the protective effect of colchicine against myocardial ischemia-reperfusion injury (I/R) and explore the underlying mechanism.
H9C2 cells exposed to hypoxia/reoxygenation (H/R) were treated with 3 nmol/L colchicine, after which the changes in cell viability were assessed using MTT assay, and AMPK phosphorylation, the expressions of NOX4, NRF2, SOD2, BAX, Bcl-2, and cleaved caspase-3 were detected with Western blotting. Male C57BL/6 mice were randomized into sham operation, I/R, I/R+colchicine, and I/R+colchicine+dorsomorphin (DSMP) groups. After the treatments, myocardial expressions of p-AMPK/AMPK, 8-OHdG, cleaved caspase-3, mitochondrial BAX (Mito-BAX), and cytoplasmic cytochrome C (Cyt-Cyto C) were examined and cardiac functions, infarct area, ATP content, and serum levels of lactic dehydrogenase (LDH) and cardiac troponin T (cTnT) levels were assessed.
In H9C2 cells, H/R exposure significantly reduced AMPK phosphorylation and expressions of NRF2, SOD2, and Bcl-2, lowered cell viability, and up-regulated the expressions of NOX4, BAX, and cleaved caspase-3 ( < 0.05), and these changes were obviously alleviated by colchicine treatment ( < 0.05). In the mouse models, myocardial I/R injury significantly reduced myocardial AMPK phosphorylation level, ATP content, and expressions of NRF2, SOD2 and Bcl-2, caused cardiac function impairment, enhanced NOX4, Mito-BAX, Cyt-Cyto C, BAX, 8-OHdG, and cleaved caspase-3 expressions, and increased infarct area and serum LDH and cTnT levels ( < 0.05). Colchicine treatment significantly reversed the damaging effects of I/R ( < 0.05), but its protective effects was obviously antagonized by DSMP ( < 0.05).
Colchicine alleviates myocardial I/R injury and protects cardiac function in mice by reducing myocardial oxidative stress and apoptosis via activating AMPK.
探讨秋水仙碱对心肌缺血再灌注损伤（I/R）的保护作用，并探究其潜在机制。
将暴露于缺氧/复氧（H/R）的H9C2细胞用3 nmol/L秋水仙碱处理，之后使用MTT法评估细胞活力变化，并用蛋白质免疫印迹法检测AMPK磷酸化、NOX4、NRF2、SOD2、BAX、Bcl-2及裂解的caspase-3的表达。将雄性C57BL/6小鼠随机分为假手术组、I/R组、I/R+秋水仙碱组及I/R+秋水仙碱+ Dorsomorphin（DSMP）组。处理后，检测心肌中p-AMPK/AMPK、8-羟基脱氧鸟苷（8-OHdG）、裂解的caspase-3、线粒体BAX（Mito-BAX）及细胞质细胞色素C（Cyt-Cyto C）的表达，并评估心脏功能、梗死面积、ATP含量以及血清乳酸脱氢酶（LDH）和心肌肌钙蛋白T（cTnT）水平。
在H9C2细胞中，H/R暴露显著降低AMPK磷酸化以及NRF2、SOD2和Bcl-2的表达，降低细胞活力，并上调NOX4、BAX及裂解的caspase-3的表达（P<0.05），而秋水仙碱处理可明显减轻这些变化（P<0.05）。在小鼠模型中，心肌I/R损伤显著降低心肌AMPK磷酸化水平、ATP含量以及NRF2、SOD2和Bcl-2的表达，导致心脏功能受损，增强NOX4、Mito-BAX、Cyt-Cyto C、BAX、8-OHdG及裂解的caspase-3的表达，并增加梗死面积以及血清LDH和cTnT水平（P<0.05）。秋水仙碱处理显著逆转I/R的损伤作用（P<0.05），但其保护作用明显被DSMP拮抗（P<0.05）。
秋水仙碱通过激活AMPK减轻小鼠心肌I/R损伤并保护心脏功能，其机制可能与减轻心肌氧化应激和细胞凋亡有关。
