Doxorubicin (DOX)-induced cardiotoxicity limits the application of DOX in cancer patients. Currently, there is no effective prevention or treatment for DOX-induced cardiotoxicity. The cellular repressor of E1A-stimulated genes (CREG1) is a cardioprotective factor that plays an important role in the maintenance of cardiomyocytes differentiation and homeostasis. However, the role and mechanism of CREG1 in DOX-induced cardiotoxicity has not yet been elucidated. , C57BL/6J mice, CREG1 transgenic and cardiac-specific CREG1 knockout mice were used to establish a DOX-induced cardiotoxicity model. H&E staining, Masson’s trichrome, WGA staining, real-time PCR, and western blotting were performed to examine fibrosis and ferroptosis in the myocardium. , neonatal mouse cardiomyocytes (NMCMs) were cultured and stimulated with DOX, CREG1-overexpressed adenovirus, and small interfering RNA was used to establish CREG1 overexpression or knockdown cardiomyocytes. Transcriptomics, real-time PCR, western blotting, and immunoprecipitation were used to examine the roles and mechanisms of CREG1 in cardiomyocytes ferroptosis. The mRNA and protein levels of CREG1 were reduced in the hearts and NMCMs after DOX treatment. CREG1 overexpression alleviated myocardial damage and inhibited DOX-induced ferroptosis in the myocardium. CREG1 deficiency in the heart aggravated DOX-induced cardiotoxicity and ferroptosis. , CREG1 overexpression inhibited cardiomyocytes ferroptosis induced by DOX, and CREG1 knockdown aggravated DOX-induced cardiotoxicity. Mechanistically, CREG1 inhibited the mRNA and protein expression of pyruvate dehydrogenase kinase 4 (PDK4) by regulating the F-box and WD repeat domain containing 7 (FBXW7)-forkhead box O1 (FOXO1) pathway. PDK4 deficiency reversed the effects of CREG1 knockdown on cardiomyocytes ferroptosis following DOX treatment. CREG1 alleviated DOX-induced cardiotoxicity by inhibiting ferroptosis in cardiomyocytes. Our findings may help clarify the new roles of CREG1 in the development of DOX-induced cardiotoxicity.

中文翻译：

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CREG1通过抑制心肌细胞铁死亡来减轻阿霉素诱导的心脏毒性


阿霉素 (DOX) 引起的心脏毒性限制了 DOX 在癌症患者中的应用。目前，DOX引起的心脏毒性尚无有效的预防或治疗方法。 E1A刺激基因的细胞阻遏物（CREG1）是一种心脏保护因子，在维持心肌细胞分化和稳态中发挥重要作用。然而，CREG1在DOX诱导的心脏毒性中的作用和机制尚未阐明。采用C57BL/6J小鼠、CREG1转基因小鼠和心脏特异性CREG1敲除小鼠建立DOX诱导的心脏毒性模型。进行H&E染色、Masson三色、WGA染色、实时PCR和蛋白质印迹来检查心肌中的纤维化和铁死亡。 ，用DOX、CREG1过表达的腺病毒培养和刺激新生小鼠心肌细胞（NMCM），并使用小干扰RNA建立CREG1过表达或敲低的心肌细胞。采用转录组学、实时 PCR、蛋白质印迹和免疫沉淀来研究 CREG1 在心肌细胞铁死亡中的作用和机制。 DOX 处理后，心脏和 NMCM 中 CREG1 的 mRNA 和蛋白水平降低。 CREG1过表达可减轻心肌损伤并抑制DOX诱导的心肌铁死亡。心脏中 CREG1 缺乏会加重 DOX 诱导的心脏毒性和铁死亡。 ，CREG1过表达抑制DOX诱导的心肌细胞铁死亡，CREG1敲低加剧了DOX诱导的心脏毒性。从机制上讲，CREG1通过调节F-box和WD重复结构域含有7 (FBXW7)-forkhead box O1 (FOXO1)途径抑制丙酮酸脱氢酶激酶4 (PDK4)的mRNA和蛋白表达。 PDK4 缺陷逆转了 DOX 治疗后 CREG1 敲低对心肌细胞铁死亡的影响。 CREG1 通过抑制心肌细胞铁死亡来减轻 DOX 诱导的心脏毒性。我们的研究结果可能有助于阐明 CREG1 在 DOX 诱导的心脏毒性发展中的新作用。