Shi Huairui, Zhang Xuehong, He Zekun, Wu Zhiyong, Rao Liya, Li Yushu
Cell Physiol Biochem. 2017;41(1):413-421. doi: 10.1159/000456531. Epub 2017 Jan 27.
The migration of cardiac fibroblasts to the infarct region plays a major role in the repair process after myocardial necrosis or damage. However, few studies investigated whether early hypoxia in cardiomyocytes induces the migration of cardiac fibroblasts. The purpose of this study was to assess the role of metabolites of early hypoxic cardiomyocytes in the induction of cardiac fibroblast migration.
Neonatal rat heart tissue was digested with a mixture of trypsin and collagenase at an appropriate ratio. Cardiomyocytes and cardiac fibroblasts were cultured via differential adhesion. The cardiomyocyte cultures were subjected to hypoxia for 2, 4, 6, 8, 10, and 12 h. The supernatants of the cardiomyocyte cultures were collected to determine the differences in cardiac fibroblast migration induced by hypoxic cardiomyocyte metabolites at various time points using a Transwell apparatus. Meanwhile, ELISA was performed to measure TNF-α, IL-1β and TGF-β expression levels in the cardiomyocyte metabolites at various time points.
The metabolites of hypoxic cardiomyocytes significantly induced the migration of cardiac fibroblasts. The induction of cardiac fibroblast migration was significantly enhanced by cardiomyocyte metabolites in comparison to the control after 2, 4, and 6 h of hypoxia, and the effect was most significant after 2 h. The expression levels of TNF-α, IL-1β, IL-6, and TGF-β were substantially increased in the metabolites of cardiomyocytes, and neutralization with anti-TNF-α and anti-IL-1β antibodies markedly reduced the induction of cardiac fibroblast migration by the metabolites of hypoxic cardiomyocytes.
The metabolites of early hypoxic cardiomyocytes can induce the migration of cardiac fibroblasts, and TNF-α and IL-1β may act as the initial chemotactic inducers.
心肌成纤维细胞向梗死区域的迁移在心肌坏死或损伤后的修复过程中起主要作用。然而，很少有研究调查心肌细胞早期缺氧是否会诱导心肌成纤维细胞的迁移。本研究的目的是评估早期缺氧心肌细胞的代谢产物在诱导心肌成纤维细胞迁移中的作用。
用胰蛋白酶和胶原酶按适当比例混合消化新生大鼠心脏组织。通过差异贴壁法培养心肌细胞和心肌成纤维细胞。将心肌细胞培养物进行2、4、6、8、10和12小时的缺氧处理。收集心肌细胞培养物的上清液，使用Transwell小室测定不同时间点缺氧心肌细胞代谢产物诱导的心肌成纤维细胞迁移差异。同时，进行ELISA检测不同时间点心肌细胞代谢产物中TNF-α、IL-1β和TGF-β的表达水平。
缺氧心肌细胞的代谢产物显著诱导心肌成纤维细胞的迁移。与对照组相比，缺氧2、4和6小时后，心肌细胞代谢产物对心肌成纤维细胞迁移的诱导作用显著增强，2小时后效果最为显著。心肌细胞代谢产物中TNF-α、IL-1β、IL-6和TGF-β的表达水平大幅升高，用抗TNF-α和抗IL-1β抗体中和后，显著降低了缺氧心肌细胞代谢产物对心肌成纤维细胞迁移的诱导作用。
早期缺氧心肌细胞的代谢产物可诱导心肌成纤维细胞的迁移，TNF-α和IL-1β可能作为初始趋化诱导因子。
