环状 RNA 00091761 的下调通过 microRNA-335-3p/ASCL4 轴防止心肌梗死后心力衰竭。

Downregulation of circular RNA 00091761 protects against heart failure after myocardial infarction via microRNA-335-3p/ ASCL4 axis.

机构信息

Department of Ultrasonics Medicine, Chengdu Medical College, Clinical Medical College and The First Affiliated Hospital of Chengdu Medical College, Sichuan 610500 China.

Department of Cardiovascular Medicine, Chengdu Medical College, Clinical Medical College and The First Affiliated Hospital of Chengdu Medical College, Sichuan 610500 China.

出版信息

Acta Biochim Pol. 2023 Sep 6;70(3):509-516. doi: 10.18388/abp.2020_6404.

DOI:10.18388/abp.2020_6404

PMID:37672718

Abstract

Our research tended to explore the biological roles and expression status of circ_00091761 in HF after MI. The hypoxia reoxygenation (H/R) injured H9c2 cells model was constructed to simulate HF after MI. The expression of circ_0091761 was examined in H/R injured H9c2 cells by qRT-PCR. Then, the effect of circ_0091761 expression on the proliferation of H/R injured H9c2 cells was evaluated by CCK-8 along with TUNEL assay. Secretion of lactate dehydrogenase (LDH), reactive oxygen species (ROS), Fe2+, glutathione (GSH), and malondialdehyde (MDA) was measured to evaluate cell ferroptosis of H/R injured H9c2 cells, along with protein levels of glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), and transferrin receptor protein (TFRC). Luciferase reporter as well as RNA pull-down assays revealed the binding relationship between miR-335-3p and circ_0091761 or ASCL4. Circ_0091761 was upregulated in H/R injured H9c2 cells. Knockdown of circ_0091761 promoted cell proliferation and suppressed ferroptosis of H/R injured H9c2 cells. Interestingly, circ_0091761 sponges miR-335-3p to upregulate acyl-CoA synthetase long-chain family member 4 (ACSL4) expression. miR-335-3p inhibitor attenuated the effects of circ_0091761 knockdown on cell proliferation and ferroptosis in H/R injured H9c2 cells. Additionally, upregulated ACSL4 abrogated elevated miR-335-3p-induced effects on H/R injured H9c2 cells. Circ_0091761 inhibited cell proliferation and accelerated ferroptosis of H/R injured H9c2 cells by sponging miR-335-3p to upregulated TFRC axis. Therefore, Inhibition of circ_0091761 may protect against HF after MI.

摘要

我们的研究倾向于探索 circ_00091761 在 MI 后 HF 中的生物学作用和表达状态。构建缺氧复氧（H/R）损伤的 H9c2 细胞模型模拟 MI 后 HF。通过 qRT-PCR 检测 H/R 损伤的 H9c2 细胞中 circ_0091761 的表达。然后，通过 CCK-8 联合 TUNEL 测定评估 circ_0091761 表达对 H/R 损伤的 H9c2 细胞增殖的影响。测量乳酸脱氢酶（LDH）、活性氧（ROS）、Fe2+、谷胱甘肽（GSH）和丙二醛（MDA）的分泌，以评估 H/R 损伤的 H9c2 细胞的细胞铁死亡，并检测谷胱甘肽过氧化物酶 4（GPX4）、溶质载体家族 7 成员 11（SLC7A11）和转铁蛋白受体蛋白（TFRC）的蛋白水平。荧光素酶报告和 RNA 下拉测定揭示了 miR-335-3p 与 circ_0091761 或 ASCL4 的结合关系。Circ_0091761 在 H/R 损伤的 H9c2 细胞中上调。Circ_0091761 敲低促进 H/R 损伤的 H9c2 细胞增殖并抑制铁死亡。有趣的是，circ_0091761 海绵 miR-335-3p 上调酰基辅酶 A 合成酶长链家族成员 4（ACSL4）的表达。miR-335-3p 抑制剂减弱了 circ_0091761 敲低对 H/R 损伤的 H9c2 细胞增殖和铁死亡的影响。此外，上调的 ACSL4 消除了 miR-335-3p 诱导的对 H/R 损伤的 H9c2 细胞的升高作用。Circ_0091761 通过海绵 miR-335-3p 抑制 TFRC 轴来抑制 H/R 损伤的 H9c2 细胞增殖并加速铁死亡。因此，抑制 circ_0091761 可能防止 MI 后 HF。