Qiu Li, Huang Zhaoming
Department of Rehabilitation Science, Faculty of Education, East China Normal University, Shanghai, China.
Cell Biochem Biophys. 2025 Jun;83(2):1565-1573. doi: 10.1007/s12013-024-01567-4. Epub 2024 Sep 30.
Pediatric pneumonia is an inflammatory disease with a very high incidence. IGF binding protein 7 (IGFBP7) plays an important role in inflammatory diseases. However, the role of IGFBP7 in pediatric pneumonia and its mechanism have not been reported. Human embryonic lung (WI-38) cells were induced by lipopolysaccharide (LPS) to construct the cell inflammatory injury model. Subsequently, the expression of IGFBP7 was detected by qPCR and western blot. Next, IGFBP7 interference plasmid was constructed, and cell viability and apoptosis were detected by CCK8, flow cytometry and western blot. ELISA and other techniques were used to detect the inflammatory level. Autophagy and mitochondrial activities were detected by immunofluorescence and other techniques, and mitophagy-related proteins were detected by western blot. To further investigate the regulatory mechanism of IGFBP7, we administered cyclosporin A, a mitophagy inhibitor, and then detected apoptosis and inflammation. The expression of IGFBP7 was significantly increased in LPS-induced WI-38 cells. Interference with IGFBP7 expression in LPS-induced cells significantly increased cell activity, decreased apoptosis and cellular inflammation levels. During this process, mitophagy was enhanced. Further addition of cyclosporin A significantly reversed the protective effect of IGFBP7 knockdown. To be concluded, inhibition of IGFBP7 alleviates LPS-induced inflammation and apoptosis in WI-38 cells via enhancing mitophagy.
小儿肺炎是一种发病率极高的炎症性疾病。胰岛素样生长因子结合蛋白7（IGFBP7）在炎症性疾病中发挥重要作用。然而，IGFBP7在小儿肺炎中的作用及其机制尚未见报道。用脂多糖（LPS）诱导人胚肺（WI-38）细胞构建细胞炎症损伤模型。随后，通过qPCR和蛋白质免疫印迹法检测IGFBP7的表达。接下来，构建IGFBP7干扰质粒，并通过CCK8、流式细胞术和蛋白质免疫印迹法检测细胞活力和凋亡情况。采用酶联免疫吸附测定（ELISA）等技术检测炎症水平。通过免疫荧光等技术检测自噬和线粒体活性，并通过蛋白质免疫印迹法检测线粒体自噬相关蛋白。为进一步研究IGFBP7的调控机制，我们给予线粒体自噬抑制剂环孢素A，然后检测细胞凋亡和炎症情况。在LPS诱导的WI-38细胞中，IGFBP7的表达显著增加。干扰LPS诱导细胞中IGFBP7的表达可显著提高细胞活性，降低细胞凋亡和细胞炎症水平。在此过程中，线粒体自噬增强。进一步添加环孢素A可显著逆转IGFBP7基因敲低的保护作用。综上所述，抑制IGFBP7可通过增强线粒体自噬减轻LPS诱导的WI-38细胞炎症和凋亡。
