心脏纤维化是心血管疾病的常见病理特征,是由于成纤维细胞的过度活化和细胞外基质(ECM)的过度沉积,导致心脏功能受损和潜在的心力衰竭或心律失常。心肌细胞(CMs)释放的细胞外囊泡(EV)调节心肌稳态所必需的各种生理功能,在心脏病中受到破坏。因此,健康的CM衍生的EV代表了一种有前途的无细胞治疗心脏纤维化。为此,我们优化了成年CM的培养条件,通过使用确定的小分子组合,在不损害细胞完整性的前提下,获得了大量的EV.电动汽车通过超速离心分离,并对其特点进行了分析。最后,测试了它们对纤维化的影响。使用我们的培养系统用源自CM的EV处理TGFβ活化的人心脏成纤维细胞导致成纤维细胞活化标志物和ECM积累的减少。获救表型与特定的EV货物相关,包括多种肌细胞特异性和抗纤维化microRNAs,尽管它们单独的效果不如EV治疗有效。值得注意的是,通路分析显示,EV治疗能逆转活化成纤维细胞的转录,并降低了几个信号通路,包括MAPK,mTOR,JAK/STAT,TGFβ,和PI3K/Akt,所有这些都与纤维化发展有关。在心脏纤维化动物模型中心内注射CM衍生的EV减少了纤维化面积并增加了血管生成,这与心脏功能的改善有关。这些发现表明,源自人类成人CM的电动汽车可能为心脏纤维化提供有针对性的有效治疗方法。由于它们的抗纤维化特性和货物的特异性。
Cardiac fibrosis is a common pathological feature of cardiovascular diseases that arises from the hyperactivation of fibroblasts and excessive extracellular matrix (ECM) deposition, leading to impaired cardiac function and potentially heart failure or arrhythmia. Extracellular vesicles (
EVs) released by cardiomyocytes (CMs) regulate various physiological functions essential for myocardial homeostasis, which are disrupted in cardiac disease. Therefore, healthy CM-derived
EVs represent a promising cell-free therapy for the treatment of cardiac fibrosis. To this end, we optimized the culture conditions of human adult CMs to obtain a large yield of EVs without compromising cellular integrity by using a defined combination of small molecules.
EVs were isolated by ultracentrifugation, and their characteristics were analysed. Finally, their effect on fibrosis was tested. Treatment of TGFβ-activated human cardiac fibroblasts with
EVs derived from CMs using our culture system resulted in a decrease in fibroblast activation markers and ECM accumulation. The rescued phenotype was associated with specific EV cargo, including multiple myocyte-specific and antifibrotic microRNAs, although their effect individually was not as effective as the EV treatment. Notably, pathway analysis showed that EV treatment reverted the transcription of activated fibroblasts and decreased several signalling pathways, including MAPK, mTOR, JAK/STAT, TGFβ, and PI3K/Akt, all of which are involved in fibrosis development. Intracardiac injection of CM-derived
EVs in an animal model of cardiac fibrosis reduced fibrotic area and increased angiogenesis, which correlated with improved cardiac function. These findings suggest that EVs derived from human adult CMs may offer a targeted and effective treatment for cardiac fibrosis, owing to their antifibrotic properties and the specificity of cargo.