罗氟司特通过靶向 SIRT1 减轻蒽环类药物诱导的心肌细胞炎症和衰老
Roflumilast Attenuates Doxorubicin-Induced Cardiotoxicity by Targeting Inflammation and Cellular Senescence in Cardiomyocytes Mediated by SIRT1.
机构信息
Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, People's Republic of China.
Department of Cardiology, The Third Affiliated Hospital of Soochow University, Changzhou, Jiangsu 213004, People's Republic of China.
出版信息
Drug Des Devel Ther. 2021 Jan 11;15:87-97. doi: 10.2147/DDDT.S269029. eCollection 2021.
BACKGROUND AND PURPOSE
Cardiotoxicity is an important side effect of the treatment of a malignant tumor with Doxorubicin. Currently, decreasing the dosage of Doxorubicin to alleviate the side effects on cardiac function is the common method to deal with the cardiotoxicity induced by Doxorubicin. The present study aims to investigate the therapeutic effects of Roflumilast on Doxorubicin-induced inflammation and cellular senescence, as well as the potential mechanism in H9c2 myocardial cells.
METHODS
The injured cardiac cell model was established by incubation with 5 μmol/L Doxorubicin. MTT was used to evaluate the cell viability of treated H9c2 cardiac cells. The expression of 4-HNE was determined using an immunofluorescence assay. The gene expression levels of IL-17, IL-6, TNF-α, IL-4, PAI-1, p21, and SIRT1 were evaluated using qRT-PCR and the protein levels of Gpx4, PAI-1, p21, and SIRT1 were determined using Western blot analysis. Secretions of IL-17, IL-6, TNF-α, IL-4, CK-MB, and cTnI were measured using ELISA. Cellular senescence was assessed using SA-β-Gal staining. Si-RNA technology was used to knockdown the expression of SIRT1 in H9c2 cardiac cells.
RESULTS
Cell viability of H9c2 cardiac cells was significantly inhibited by Doxorubicin but rescued by Roflumilast. The upregulated 4-HNE and downregulated Gpx4 were reversed by Roflumilast. The secretions of IL-6 and IL-17 were promoted by Doxorubicin and suppressed by Roflumilast. The increased SA-β-Gal staining induced by Doxorubicin was inhibited by Roflumilast. P21 and PAI-1 were significantly upregulated and SIRT1 was greatly downregulated by Doxorubicin, all of which were reversed by Roflumilast. The anti-senescent effect of Roflumilast was abolished by knocking down SIRT1.
CONCLUSION
Roflumilast might attenuate Doxorubicin-induced inflammation and cellular senescence in cardiomyocytes by upregulating SIRT1.
背景与目的
多柔比星治疗恶性肿瘤会导致心脏毒性，这是一种重要的副作用。目前，减少多柔比星的剂量以减轻对心脏功能的副作用是应对多柔比星引起的心脏毒性的常用方法。本研究旨在探讨罗氟司特对多柔比星诱导的炎症和细胞衰老的治疗作用，以及其在 H9c2 心肌细胞中的潜在机制。
方法
用 5 μmol/L 多柔比星孵育损伤心肌细胞模型。MTT 法评估处理后的 H9c2 心肌细胞的细胞活力。用免疫荧光法测定 4-HNE 的表达。用 qRT-PCR 测定 IL-17、IL-6、TNF-α、IL-4、PAI-1、p21 和 SIRT1 的基因表达水平，用 Western blot 分析测定 Gpx4、PAI-1、p21 和 SIRT1 的蛋白水平。用 ELISA 法测定 IL-17、IL-6、TNF-α、IL-4、CK-MB 和 cTnI 的分泌。用 SA-β-半乳糖苷染色法评估细胞衰老。用 siRNA 技术敲低 H9c2 心肌细胞中的 SIRT1 表达。
结果
多柔比星显著抑制 H9c2 心肌细胞的活力，但罗氟司特可挽救其活力。罗氟司特逆转了多柔比星上调的 4-HNE 和下调的 Gpx4。多柔比星促进了 IL-6 和 IL-17 的分泌，罗氟司特抑制了其分泌。多柔比星诱导的 SA-β-半乳糖苷染色增加被罗氟司特抑制。多柔比星显著上调 p21 和 PAI-1，显著下调 SIRT1，罗氟司特均逆转了这一作用。敲低 SIRT1 可消除罗氟司特的抗衰老作用。
结论
罗氟司特可能通过上调 SIRT1 来减轻多柔比星诱导的心肌细胞炎症和衰老。
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