Zhang Jishou, Ding Wen, Yin Zheng, Liu Siqi, Zhao Mengmeng, Xu Yao, Liu Jianfang, Pan Wei, Peng Shanshan, Wei Cheng, Zheng Zihui, Qin Juan-Juan, Wan Jun, Wang Menglong
Department of Cardiology, Renmin Hospital of Wuhan University, 238 Jiefang Road, Wuhan 430060, China.
Department of Cardiology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Cardiovasc Res. 2024 Dec 14;120(16):2117-2133. doi: 10.1093/cvr/cvae208.
Interleukin (IL)-12p40 is a common subunit of the bioactive cytokines IL-12 and IL-23, and it also has its own intrinsic functional activity. However, its role in doxorubicin-induced chronic cardiomyopathy (DICCM) as well as the underlying mechanisms are still unknown.
In this study, we used IL-12p40-knockout mice, IL-23p19-knockout mice, Rag1-knockout mice, a ferroptosis inhibitor, recombinant IL-12 (rIL-12), rIL-23, rIL-12p40, rIL-12p80, and anti-IL17A to investigate the effects of IL-12p40 on DICCM and elucidate the underlying mechanisms. We found that myocardial ferroptosis were increased in DICCM and that the inhibition of ferroptosis protected against DICCM. The expression of IL-12p40 was upregulated, and IL-12p40 was predominantly expressed by CD4+ T cells in the hearts of mice with DICCM. IL-12p40 knockout attenuated cardiac dysfunction, fibrosis and ferroptosis in DICCM, and similar results were observed in the context of CD4+ T cell IL-12p40 deficiency in Rag1-/- mice. Treatment with rIL-23, but not rIL-12, rIL-12p40 monomer or rIL-12p80, abolished the protective effects of IL-12p40 knockout. Moreover, rIL-23 treatment and IL-23p19 knockout exacerbated and ameliorated DICCM, respectively. IL-12p40 knockout might protect against DICCM by inhibiting Th17 differentiation and IL-17A production but not Th1, Th2 and Treg differentiation. Neutralizing IL-17A with an antibody also attenuated cardiac dysfunction, fibrosis, and ferroptosis. The IL-12p40/Th17/IL-17A axis might promote cardiomyocyte ferroptosis by activating TNF receptor-associated factor 6 (TRAF6)/mitogen-activated protein kinase (MAPK)/P53 signalling in DICCM.
Interleukin-12p40 deficiency protects against DICCM by inhibiting Th17 differentiation and the production of IL-17A, which plays critical roles in cardiomyocyte ferroptosis in DICCM via activating TRAF6/MAPK/P53 signalling. Our study may provide novel insights for the identification of therapeutic targets for treating DICCM in the clinic.
白细胞介素（IL）-12p40是生物活性细胞因子IL-12和IL-23的共同亚基，且其自身具有内在功能活性。然而，其在阿霉素诱导的慢性心肌病（DICCM）中的作用及潜在机制仍不清楚。
在本研究中，我们使用IL-12p40基因敲除小鼠、IL-23p19基因敲除小鼠、Rag1基因敲除小鼠、一种铁死亡抑制剂、重组IL-12（rIL-12）、rIL-23、rIL-12p40、rIL-12p80和抗IL17A来研究IL-12p40对DICCM的影响并阐明潜在机制。我们发现DICCM中心肌铁死亡增加，抑制铁死亡可预防DICCM。IL-12p40的表达上调，且在DICCM小鼠心脏中IL-12p40主要由CD4⁺T细胞表达。IL-12p40基因敲除减轻了DICCM中的心脏功能障碍、纤维化和铁死亡，在Rag1⁻/⁻小鼠CD4⁺T细胞IL-12p40缺乏的情况下也观察到了类似结果。用rIL-23而非rIL-12、rIL-12p40单体或rIL-12p80处理消除了IL-12p40基因敲除的保护作用。此外，rIL-23处理和IL-23p19基因敲除分别加重和改善了DICCM。IL-12p40基因敲除可能通过抑制Th17分化和IL-17A产生来预防DICCM，但不影响Th1、Th2和Treg分化。用抗体中和IL-17A也减轻了心脏功能障碍、纤维化和铁死亡。在DICCM中，IL-12p40/Th17/IL-17A轴可能通过激活肿瘤坏死因子受体相关因子6（TRAF6）/丝裂原活化蛋白激酶（MAPK）/P53信号通路促进心肌细胞铁死亡。
白细胞介素-12p40缺乏通过抑制Th17分化和IL-17A产生来预防DICCM，IL-17A在DICCM的心肌细胞铁死亡中通过激活TRAF6/MAPK/P53信号通路发挥关键作用。我们的研究可能为临床治疗DICCM的治疗靶点识别提供新的见解。
