Hao Jingxia, Wang Hua, Li Bo, Chen Jingshi, Wang Wenlu, Wang Yingxue, Zhang Yingqian
Hebei Provincial Key Laboratory of Pediatric Cardiovascular Disease, Hebei Children's Hospital, Shijiazhuang, 050031, Hebei, China.
Sci Rep. 2025 Aug 12;15(1):29522. doi: 10.1038/s41598-025-04375-9.
The purpose of this study was to investigate whether tyrosine phosphatase Shp2 can promote the progression of dilated cardiomyopathy by upregulating intracellular oxidative stress and subsequently promoting mitophagy. Macrophage-specific Shp2 knockout mice were cultured and a dilated cardiomyopathy model was established. Ultrasonography, Masson staining and WGA staining were used to detect the condition of mouse hearts. Immunofluorescence, Western blot and EL were used to detect the expression of relevant proteins in mouse myocardial tissues. Transmission electron microscopy was used to examine the morphology of mouse myocardial cells and their internal mitochondria. Primary BMMs from wild-type mice and specific Shp2 knockout mice were co-cultured with normal myocardial cell. Western blot was used to detect the expression of relevant proteins in BMMs and myocardial cells. Transwell assay was used to measure the migration and invasion ability of BMMs, and immunofluorescence staining was used to detect the expression level of PINK protein in myocardial cells to validate the Western blot results. Dilated cardiomyopathy was ameliorated in DCM mice with specific knockout of Shp2, which showed attenuation of heart structure, degree of fibrosis in myocardial tissue, and degree of cardiomyocyte dilatation. And the degree of autophagy in myocardial tissue was significantly reduced as observed by western blot and transmission electron microscopy. Subsequent cellular experiments also verified that specific knockdown of Shp2 inhibited the invasion of BMMs. And it also mediated the ROS/NF-κB/Src/FAK signalling pathway to inhibit the expression of related autophagy and apoptosis proteins. Tyrosine phosphatase Shp2 accelerated the progression of dilated cardiomyopathy via upregulating NF-κB/Src/FAK signaling induced ROS and mitophagy.
本研究的目的是探究酪氨酸磷酸酶Shp2是否可通过上调细胞内氧化应激并随后促进线粒体自噬来促进扩张型心肌病的进展。培养巨噬细胞特异性Shp2基因敲除小鼠并建立扩张型心肌病模型。使用超声心动图、Masson染色和WGA染色检测小鼠心脏状况。使用免疫荧光、蛋白质免疫印迹法(Western blot)和酶联免疫吸附测定(EL)检测小鼠心肌组织中相关蛋白的表达。使用透射电子显微镜检查小鼠心肌细胞及其内部线粒体的形态。将来自野生型小鼠和特异性Shp2基因敲除小鼠的原代骨髓巨噬细胞(BMMs)与正常心肌细胞共培养。使用蛋白质免疫印迹法检测BMMs和心肌细胞中相关蛋白的表达。使用Transwell实验测量BMMs的迁移和侵袭能力,并使用免疫荧光染色检测心肌细胞中PINK蛋白的表达水平以验证蛋白质免疫印迹法的结果。在特异性敲除Shp2的扩张型心肌病小鼠中,扩张型心肌病得到改善,表现为心脏结构、心肌组织纤维化程度和心肌细胞扩张程度减轻。并且通过蛋白质免疫印迹法和透射电子显微镜观察到心肌组织中的自噬程度显著降低。随后的细胞实验也证实,特异性敲低Shp2可抑制BMMs的侵袭。并且它还介导ROS/NF-κB/Src/FAK信号通路以抑制相关自噬和凋亡蛋白的表达。酪氨酸磷酸酶Shp2通过上调NF-κB/Src/FAK信号诱导的ROS和线粒体自噬加速扩张型心肌病的进展。