Liu Yazhong, Yin Hongshan, Wang Tao, Chen Tao, Guo Chengda, Zhang Fue, Jiang Zhian
Department of Cardiovascular Disease and Surgery, Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China.
Mol Med Rep. 2025 Jun;31(6). doi: 10.3892/mmr.2025.13520. Epub 2025 Apr 11.
The objective of the present study was to investigate the impact of myeloid Src homology region 2‑containing protein tyrosine phosphatase 2 (SHP2) on myocardial ischemia reperfusion (MI/R) injury and the underlying mechanism. Bioinformatics was used to analyze genes specifically associated with MI/R. In addition, myeloid‑specific SHP2 knockout mice and wild‑type mice were subjected to MI/R or sham surgery. Echocardiography and Masson's staining were used to observe the myocardial function and infarct area of the mice. In addition, double immunofluorescence staining was used to detect the relative fluorescence intensity of SHP2 and bromodomain‑containing protein 4 (BRD4) in bone marrow‑derived macrophages (BMMs) from the mice. Western blot analysis was conducted to determine the expression levels of SHP2, BRD4, spleen tyrosine kinase (SYK), stimulator of interferon genes (STING), NADPH oxidase 4 (NOX4), NLR family pyrin domain containing 3 (NLRP3), IL‑1β and gasdermin D (GSDMD) in BMMs and mouse myocardial cells co‑cultured with the BMMs. In addition, flow cytometry was employed to assess myocardial cell apoptosis. Bioinformatics analysis revealed the downregulated expression of SHP2 and upregulated expression of BRD4 and SYK in mice with MI/R. The deletion of myeloid SHP2 aggravated MI/R injury, impaired cardiac function and increased the infarct area in mice. In addition, myeloid SHP2 deletion in BMMs promoted the expression of BRD4, SYK, STING, NOX4 and NLRP3 in BMMs, and the expression of IL‑1β and GSDMD in mouse myocardial cells co‑cultured with the BMMs. In addition, the deletion of myeloid SHP2 promoted cardiomyocyte apoptosis. These results indicate that myeloid SHP2 inhibits MI/R injury by regulating BRD4/SYK/STING/NOX4/NLRP3 signaling in BMMs.
本研究的目的是探讨髓系含Src同源区2的蛋白酪氨酸磷酸酶2（SHP2）对心肌缺血再灌注（MI/R）损伤的影响及其潜在机制。利用生物信息学分析与MI/R特异性相关的基因。此外，将髓系特异性SHP2基因敲除小鼠和野生型小鼠进行MI/R或假手术。采用超声心动图和Masson染色观察小鼠心肌功能和梗死面积。此外，采用双重免疫荧光染色检测小鼠骨髓来源巨噬细胞（BMMs）中SHP2和含溴结构域蛋白4（BRD4）的相对荧光强度。进行蛋白质免疫印迹分析以确定BMMs和与BMMs共培养的小鼠心肌细胞中SHP2、BRD4、脾酪氨酸激酶（SYK）、干扰素基因刺激因子（STING）、NADPH氧化酶4（NOX4）、NLR家族含pyrin结构域3（NLRP3）、IL-1β和gasdermin D（GSDMD）的表达水平。此外，采用流式细胞术评估心肌细胞凋亡。生物信息学分析显示，MI/R小鼠中SHP2表达下调，BRD4和SYK表达上调。髓系SHP2的缺失加重了MI/R损伤，损害了心脏功能，增加了小鼠的梗死面积。此外，BMMs中髓系SHP2的缺失促进了BMMs中BRD4、SYK、STING、NOX4和NLRP3的表达，以及与BMMs共培养的小鼠心肌细胞中IL-1β和GSDMD的表达。此外，髓系SHP2的缺失促进了心肌细胞凋亡。这些结果表明，髓系SHP2通过调节BMMs中的BRD4/SYK/STING/NOX4/NLRP3信号通路抑制MI/R损伤。
