Department of Cardiology, Fujian Provincial Hospital, Fuzhou, Fujian 350001, P.R. China.
Department of Cardiology, The 900th Hospital of Joint Logistics Support Force of People's Liberation Army, Fuzhou, Fujian 350025, P.R. China.
Mol Med Rep. 2021 Feb;23(2). doi: 10.3892/mmr.2020.11805. Epub 2020 Dec 23.
Myocardial ischemia/reperfusion (I/R) injury is a serious complication of reperfusion therapy for myocardial infarction. At present, there is not an effective treatment strategy available for myocardial I/R. The present study aimed to investigate the effects of human tissue kallikrein 1 (hTK1) and human tissue inhibitors of matrix metalloproteinase 1 (hTIMP1) gene co‑expression on myocardial I/R injury. A rat model of myocardial I/R injury and a cell model with hypoxia/reoxygenation (H/R) treatment in cardiac microvascular endothelial cells (CMVECs) were established, and treated with adenovirus (Ad)‑hTK1/hTIMP1. Following which, histological and triphenyl‑tetrazolium‑chloride staining assays were performed. Cardiac function was tested by echocardiographic measurement. The serum levels of oxidative stress biomarkers in rats and the intracellular reactive oxygen species (ROS) levels in CMVECs were measured. Additionally, experiments, including immunostaining, reverse transcription‑quantitative PCR, western blotting, and MTT, wound healing, Transwell and tube formation assays were also performed. The results of the present study demonstrated that Ad‑hTK1/hTIMP1 alleviated myocardial injury and improved cardiac function in myocardial I/R model rats. Ad‑hTK1/hTIMP1 also significantly enhanced microvessel formation, decreased matrix metalloproteinase (MMP)2 and MMP9 expression, and reduced oxidative stress in myocardial I/R model rats. Furthermore, Ad‑hTK1/hTIMP1 significantly enhanced proliferation, migration and tube formation in H/R‑treated CMVECs. Additionally, Ad‑hTK1/hTIMP1 significantly decreased intracellular ROS production and γ‑H2A.X variant histone expression levels in H/R‑treated CMVECs. In conclusion, the results of the present study demonstrated that co‑expression of hTK1 and hTIMP1 genes displayed significant protective effects on myocardial I/R injury by promoting angiogenesis and suppressing oxidative stress; therefore, co‑expression of hTK1 and hTIMP1 may serve as a potential therapeutic strategy for myocardial I/R injury.
心肌缺血/再灌注（I/R）损伤是心肌梗死再灌注治疗的严重并发症。目前，尚无有效的心肌 I/R 治疗策略。本研究旨在探讨人组织激肽释放酶 1（hTK1）和人组织金属蛋白酶抑制剂 1（hTIMP1）基因共表达对心肌 I/R 损伤的影响。建立大鼠心肌 I/R 损伤模型和心肌微血管内皮细胞（CMVECs）缺氧/复氧（H/R）处理细胞模型，并给予腺病毒（Ad）-hTK1/hTIMP1 处理。然后进行组织学和三苯基四氮唑氯化物染色检测。通过超声心动图测量评估心功能。检测大鼠血清氧化应激生物标志物水平和 CMVECs 内活性氧（ROS）水平。此外，还进行了免疫染色、逆转录-定量 PCR、western blot 以及 MTT、划痕愈合、Transwell 和管形成实验。本研究结果表明，Ad-hTK1/hTIMP1 减轻了心肌 I/R 模型大鼠的心肌损伤，改善了心功能。Ad-hTK1/hTIMP1 还显著增强了微血管形成，降低了心肌 I/R 模型大鼠基质金属蛋白酶（MMP）2 和 MMP9 的表达，减轻了氧化应激。此外，Ad-hTK1/hTIMP1 显著增强了 H/R 处理的 CMVECs 的增殖、迁移和管形成。此外，Ad-hTK1/hTIMP1 显著降低了 H/R 处理的 CMVECs 内的 ROS 产生和 γ-H2A.X 变异组蛋白表达水平。综上所述，本研究结果表明，hTK1 和 hTIMP1 基因的共表达通过促进血管生成和抑制氧化应激对心肌 I/R 损伤具有显著的保护作用；因此，hTK1 和 hTIMP1 的共表达可能成为心肌 I/R 损伤的一种潜在治疗策略。
