Cardiovascular Research Group, Department of Biochemistry, University of Szeged, Szeged H-6720, Hungary; Pharmahungary Group, Szeged H-6722, Hungary.
MTA-SE System Pharmacology Research Group, Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest H-1089, Hungary.
Vascul Pharmacol. 2023 Oct;152:107208. doi: 10.1016/j.vph.2023.107208. Epub 2023 Aug 11.
Here we aimed to establish an in vitro engineered heart tissue (EHT) co-morbidity mimicking model of ischemia-reperfusion injury and diabetes. EHTs were generated from primary neonatal rat cardiomyocytes. Hyperglycemic conditions or hyperosmolar controls were applied for one day to model acute hyperglycemia and for seven days to model chronic hyperglycemia. 120 min' simulated ischemia (SI) was followed by 120 min' reperfusion (R) and 1-day follow-up reperfusion (FR). Normoxic controls (N) were not subjected to SI/R. Half of the EHTs was paced, the other half was left unpaced. To assess cell injury, lactate-dehydrogenase (LDH) concentration was measured. Beating force and activity (frequency) were monitored as cardiomyocyte functional parameters. LDH-release indicated relevant cell injury after SI/N in each experimental condition, with much higher effects in the chronically hyperglycemic/hyperosmolar groups. SI stopped beating of EHTs in each condition, which returned during reperfusion, with weaker recovery in chronic conditions than in acute conditions. Acutely treated EHTs showed small LDH-release and ∼80% recovery of force during reperfusion and follow-up, while chronically treated EHTs showed a marked LDH-release, only ∼30% recovery with reperfusion and complete loss of beating activity during 24 h follow-up reperfusion. We conclude that EHTs respond differently to SI/R injury in acute and chronic hyperglycemia/hyperosmolarity, and that our EHT model is a novel in vitro combination of diabetes and ischemia-reperfusion.
在这里，我们旨在建立一种体外工程心脏组织（EHT）共患病模拟模型，以模拟缺血再灌注损伤和糖尿病。EHT 由原代新生大鼠心肌细胞生成。高血糖条件或高渗对照用于模拟急性高血糖一天，模拟慢性高血糖七天。120 分钟的模拟缺血（SI）后进行 120 分钟的再灌注（R）和 1 天的再灌注随访（FR）。正常氧合对照（N）不进行 SI/R。EHT 的一半进行起搏，另一半不起搏。为了评估细胞损伤，测量乳酸脱氢酶（LDH）浓度。作为心肌细胞功能参数，监测收缩力和活性（频率）。在每个实验条件下，SI/N 后 LDH 释放表明存在相关的细胞损伤，在慢性高血糖/高渗组中的影响更大。SI 停止了每种条件下的 EHT 跳动，在再灌注期间恢复，慢性条件下的恢复比急性条件下弱。在急性处理的 EHT 中，再灌注和随访期间 LDH 释放量小，收缩力恢复约 80%，而在慢性处理的 EHT 中，LDH 释放明显，再灌注时仅恢复约 30%，24 小时随访再灌注期间完全丧失跳动活性。我们得出结论，EHT 对急性和慢性高血糖/高渗性缺血再灌注损伤的反应不同，我们的 EHT 模型是糖尿病和缺血再灌注的新型体外组合。
