Abstract：
UNASSIGNED: The progression and persistence of myocardial ischemia/reperfusion injury (MI/RI) are strongly linked to local inflammatory responses and oxidative stress. Cyclophilin A (CypA), a pro-inflammatory factor, is involved in various cardiovascular diseases. However, the role and mechanism of action of CypA in MI/RI are still not fully understood.
UNASSIGNED: We used the Gene Expression Omnibus (GEO) database for bioinformatic analysis. We collected blood samples from patients and controls for detecting the levels of serum CypA using enzyme-linked immunosorbent assay (ELISA) kits. We then developed a myocardial ischemia/reperfusion (I/R) injury model in wild-type (WT) mice and Ppia-/- mice. We utilized echocardiography, hemodynamic measurements, hematoxylin and eosin (H&E) staining, immunohistochemistry, enzyme-linked immunosorbent assay, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining to determine the role of CypA in myocardial I/R injury. Finally, we conducted an in vitrostudy, cell transfection, flow cytometry, RNA interference, and a co-immunoprecipitation assay to clarify the mechanism of CypA in aggravating cardiomyocyte apoptosis.
UNASSIGNED: We found that CypA inhibited TXNIP degradation to enhance oxidative stress-induced cardiomyocyte apoptosis during MI/RI. By comparing and analyzing CypA expression in patients with coronary atherosclerotic heart disease and in healthy controls, we found that CypA was upregulated in patients with Coronary Atmospheric Heart Disease, and its expression was positively correlated with Gensini scores. In addition, CypA deficiency decreased cytokine expression, oxidative stress, and cardiomyocyte apoptosis in I/R-treated mice, eventually alleviating cardiac dysfunction. CypA knockdown also reduced H2O2-induced apoptosis in H9c2 cells. Mechanistically, we found that CypA inhibited K48-linked ubiquitination mediated by atrophin-interacting protein 4 (AIP4) and proteasomal degradation of TXNIP, a thioredoxin-binding protein that mediates oxidative stress and induces apoptosis.
UNASSIGNED: These findings highlight the critical role CypA plays in myocardial injury caused by oxidative stress-induced apoptosis, indicating that CypA can be a viable biomarker and a therapeutic target candidate for MI/RI.
UNASSIGNED: We used the Gene Expression Omnibus (GEO) database for bioinformatic analysis. We collected blood samples from patients and controls for detecting the levels of serum CypA using enzyme-linked immunosorbent assay (ELISA) kits. We then developed a myocardial ischemia/reperfusion (I/R) injury model in wild-type (WT) mice and Ppia-/- mice. We utilized echocardiography, hemodynamic measurements, hematoxylin and eosin (H&E) staining, immunohistochemistry, enzyme-linked immunosorbent assay, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining to determine the role of CypA in myocardial I/R injury. Finally, we conducted an in vitrostudy, cell transfection, flow cytometry, RNA interference, and a co-immunoprecipitation assay to clarify the mechanism of CypA in aggravating cardiomyocyte apoptosis.
UNASSIGNED: We found that CypA inhibited TXNIP degradation to enhance oxidative stress-induced cardiomyocyte apoptosis during MI/RI. By comparing and analyzing CypA expression in patients with coronary atherosclerotic heart disease and in healthy controls, we found that CypA was upregulated in patients with Coronary Atmospheric Heart Disease, and its expression was positively correlated with Gensini scores. In addition, CypA deficiency decreased cytokine expression, oxidative stress, and cardiomyocyte apoptosis in I/R-treated mice, eventually alleviating cardiac dysfunction. CypA knockdown also reduced H2O2-induced apoptosis in H9c2 cells. Mechanistically, we found that CypA inhibited K48-linked ubiquitination mediated by atrophin-interacting protein 4 (AIP4) and proteasomal degradation of TXNIP, a thioredoxin-binding protein that mediates oxidative stress and induces apoptosis.
UNASSIGNED: These findings highlight the critical role CypA plays in myocardial injury caused by oxidative stress-induced apoptosis, indicating that CypA can be a viable biomarker and a therapeutic target candidate for MI/RI.
摘要：
■心肌缺血/再灌注损伤（MI/RI）的进展和持续与局部炎症反应和氧化应激密切相关。亲环蛋白A(CypA),一种促炎因子，与各种心血管疾病有关。然而,CypA在MI/RI中的作用和作用机制尚不完全清楚。
■我们使用基因表达综合(GEO)数据库进行生物信息学分析。我们收集患者和对照组的血液样本，使用酶联免疫吸附测定（ELISA）试剂盒检测血清CypA水平。然后,我们在野生型(WT)小鼠和Ppia-/-小鼠中开发了心肌缺血/再灌注(I/R)损伤模型。我们用了超声心动图,血液动力学测量,苏木精和伊红（H&E）染色，免疫组织化学，酶联免疫吸附测定，和末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）染色，以确定CypA在心肌I/R损伤中的作用。最后,我们进行了一项体外研究,细胞转染，流式细胞术,RNA干扰,并采用免疫共沉淀法阐明CypA加重心肌细胞凋亡的机制。
■我们发现CypA抑制TXNIP降解以增强MI/RI期间氧化应激诱导的心肌细胞凋亡。通过比较和分析CypA在冠状动脉粥样硬化性心脏病患者和健康对照组中的表达，我们发现CypA在冠心病患者中上调，其表达与Gensini评分呈正相关。此外,CypA缺乏降低细胞因子表达，氧化应激，I/R处理小鼠心肌细胞凋亡，最终缓解心功能不全。CypA敲低也减少H2O2诱导的H9c2细胞凋亡。机械上,我们发现CypA抑制了由atrophin相互作用蛋白4（AIP4）介导的K48连接的泛素化和TXNIP的蛋白酶体降解，一种介导氧化应激和诱导细胞凋亡的硫氧还蛋白结合蛋白。
■这些发现强调了CypA在氧化应激诱导的细胞凋亡引起的心肌损伤中的关键作用，表明CypA可以是MI/RI的可行生物标志物和治疗靶标候选物。
■我们使用基因表达综合(GEO)数据库进行生物信息学分析。我们收集患者和对照组的血液样本，使用酶联免疫吸附测定（ELISA）试剂盒检测血清CypA水平。然后,我们在野生型(WT)小鼠和Ppia-/-小鼠中开发了心肌缺血/再灌注(I/R)损伤模型。我们用了超声心动图,血液动力学测量,苏木精和伊红（H&E）染色，免疫组织化学，酶联免疫吸附测定，和末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）染色，以确定CypA在心肌I/R损伤中的作用。最后,我们进行了一项体外研究,细胞转染，流式细胞术,RNA干扰,并采用免疫共沉淀法阐明CypA加重心肌细胞凋亡的机制。
■我们发现CypA抑制TXNIP降解以增强MI/RI期间氧化应激诱导的心肌细胞凋亡。通过比较和分析CypA在冠状动脉粥样硬化性心脏病患者和健康对照组中的表达，我们发现CypA在冠心病患者中上调，其表达与Gensini评分呈正相关。此外,CypA缺乏降低细胞因子表达，氧化应激，I/R处理小鼠心肌细胞凋亡，最终缓解心功能不全。CypA敲低也减少H2O2诱导的H9c2细胞凋亡。机械上,我们发现CypA抑制了由atrophin相互作用蛋白4（AIP4）介导的K48连接的泛素化和TXNIP的蛋白酶体降解，一种介导氧化应激和诱导细胞凋亡的硫氧还蛋白结合蛋白。
■这些发现强调了CypA在氧化应激诱导的细胞凋亡引起的心肌损伤中的关键作用，表明CypA可以是MI/RI的可行生物标志物和治疗靶标候选物。
