Zhang Yuezhan, Lu Hongxiang, Guo Ting, Wang Jun
Department of Geriatrics, Lianyungang Affiliated Hospital of Nanjing University of Chinese Medicine, Lianyungang, 222004, China.
Department of Laboratory, Lianyungang Affiliated Hospital of Nanjing University of Chinese Medicine, Lianyungang, 222004, China.
Mol Neurobiol. 2025 Feb;62(2):1960-1970. doi: 10.1007/s12035-024-04379-y. Epub 2024 Jul 25.
Results of previous studies suggested that programmed cell death 4 (PDCD4) was overexpressed in cerebral ischemia (CI), and mothers against decapentaplegic homolog 1 (SMAD1) is a transcription factor of PDCD4, and it is also elevated in CI; however, the regulatory mechanism of SMAD1/PDCD4 axis in CI remains unclear. The current work has been designed to explore the role and associated mechanisms of SMAD1/PDCD4 in CI. PDCD4 and SMAD1 expressions have been examined by real-time reverse transcription-polymerase chain reaction (RT-qPCR) method, and receiver operating characteristic (ROC) curve analysis has been performed to determine the potential diagnostic value of PDCD4 and SMAD1. An oxygen-glucose deprivation (OGD) model has been used to investigate the effects of PDCD4 and SMAD1 on CI in vitro. Cell apoptosis was evaluated using TdT-mediated dUTP nick end labeling (TUNEL) assays. The interaction between SMAD1 and PDCD4 axis has been confirmed by using dual-luciferase reporter as well as chromatin immunoprecipitation (Ch-IP) assays. Finally, the effects of SMAD1/PDCD4 axis on the ferroptosis of neuron cells have been examined. PDCD4 was overexpressed in blood samples of CI patients. ROC analysis showed the AUC for PDCD4 was 0.7478, and NIHSS and MRS scores were positively correlated with PDCD4 expression. Moreover, the cellular OGD model was established and knockdown of PDCD4 suppressed the apoptosis of neurons. Besides, knockdown of PDCD4 also inhibited ferroptosis of OGD-treated neuron cells in vitro. Additionally, SMAD1 was upregulated in blood samples of CI patients, NIHSS and MRS scores were positively correlated with SMAD1 expression, and SMAD1 is a transcriptional factor of PDCD4, and SMAD1 could transcriptionally regulate the expression of PDCD4. Finally, SMAD1 could regulate the ferroptosis of neuron cells through regulating the transcription of PDCD4. The SMAD1/PDCD4 axis regulates the growth, apoptosis, and ferroptosis of neuron cells, suggesting that targeting the SMAD1/PDCD4 axis may be a potential therapeutic method.
先前研究结果表明，程序性细胞死亡4（PDCD4）在脑缺血（CI）中过表达，而母亲对五体不全同源物1（SMAD1）是PDCD4的转录因子，在CI中也升高；然而，SMAD1/PDCD4轴在CI中的调控机制仍不清楚。当前研究旨在探讨SMAD1/PDCD4在CI中的作用及相关机制。通过实时逆转录-聚合酶链反应（RT-qPCR）法检测PDCD4和SMAD1的表达，并进行受试者工作特征（ROC）曲线分析以确定PDCD4和SMAD1的潜在诊断价值。采用氧糖剥夺（OGD）模型在体外研究PDCD4和SMAD1对CI的影响。使用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）检测评估细胞凋亡。通过双荧光素酶报告基因以及染色质免疫沉淀（Ch-IP）检测证实SMAD1与PDCD4轴之间的相互作用。最后，检测SMAD1/PDCD4轴对神经元细胞铁死亡的影响。PDCD4在CI患者血液样本中过表达。ROC分析显示PDCD4的曲线下面积（AUC）为0.7478，美国国立卫生研究院卒中量表（NIHSS）和改良Rankin量表（MRS）评分与PDCD4表达呈正相关。此外，建立了细胞OGD模型，敲低PDCD4可抑制神经元凋亡。此外，敲低PDCD4还可在体外抑制OGD处理的神经元细胞的铁死亡。另外，SMAD1在CI患者血液样本中上调，NIHSS和MRS评分与SMAD1表达呈正相关，且SMAD1是PDCD4的转录因子，SMAD1可转录调控PDCD4的表达。最后，SMAD1可通过调控PDCD4的转录来调节神经元细胞的铁死亡。SMAD1/PDCD4轴调节神经元细胞的生长、凋亡和铁死亡，提示靶向SMAD1/PDCD4轴可能是一种潜在的治疗方法。
