低氧对心脏成纤维细胞增殖迁移活性的影响及作用机制

李赞; 程培培; 丰明慧; 吕嵘; 徐明

doi:10.12125/j.chj.202108022

低氧对心脏成纤维细胞增殖迁移活性的影响及作用机制

Effect of hypoxia on proliferation and migration activity of cardiac fibroblasts and its mechanism

摘要

摘要:
目的探讨低氧对乳鼠心脏成纤维细胞(cardiac fibroblasts, CFs)增殖和迁移活性的影响及其作用机制。
方法原代分离培养C57BL/6J乳鼠CFs，给予低氧（10 ml/L O₂）、常氧（210 ml/L O₂）处理，CCK-8和免疫荧光法检测CFs增殖能力；划痕实验观察CFs的迁移活性；蛋白质免疫印迹法检测相关蛋白的改变；药物阻断和基因干预关键分子观察对CFs生物活性的影响。
结果相较于常氧组，低氧可显著促进CFs的增殖、迁移活性；低氧下CFs中HIF1α 蛋白表达水平升高；应用药物特异性阻断以及siRNA抑制HIF1α 的表达均可显著降低CFs的增殖和迁移活性。
结论低氧可通过HIF1α促进CFs增殖和迁移，这可能是心脏纤维化发生的重要机制之一。

Abstract:
AIM To investigate the effect of hypoxia on the proliferation and migration activity of neonatal mouse cardiac fibroblasts (CFs) and to explore the underlying mechanism.
METHODS C57BL/6J neonatal mouse CFs were isolated, cultured and treated with hypoxia (10 ml/L O₂) and normoxia (210 ml/L O₂). CCK-8 and immunofluorescence method were used to detect the proliferation ability of CFs, light field images were captured for cell migration activity assessment, Western blotting was used to detect changes in related proteins, and drug blocking assay and gene interference were further applied to observe the underlying mechanism.
RESULTS Hypoxia treatment significantly promoted CFs proliferation and migration compared with normoxia treatment. Meanwhile, blocking HIF1α by 2-MeOE2 and small interfering RNA (siRNA) targeting HIF1α markedly blunted CFs proliferation and migration, which was in parallel with increased expression of HIF1α after CFs exposed to hypoxia.
CONCLUSION Hypoxia promotes the proliferation and migration of CFs via HIF1α, which provides a cue that oxygen deprivation triggers cellular activation of cardiac fibrosis.

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