Effect of Macrophages-derived SHP2 on the Apoptotic Phenotype of Endo-metriosis by Promoting Oxidative Stress and Angiogenesis Via the PI3K/PTEN Pathway

CAI Li; WU Ping; TANG Haixu; ZHANG Chunhua; FU Xueshu; XIONG Qian

doi:10.3969/j.issn.1002-3429.2024.16.015

Abstract：Objective To study the effect of Src homology 2-containing protein tyrosine phosphatase 2(SHP2)in promoting oxidative stress and angiogenesis through phosphatidylinositol-3-kinase(PI3K)/phosphate and tension homology de-leted on chromosome ten(PTEN)pathway on apoptotic phenotype of endometriosis.Methods The ectopic endometrial tis-sues of 50 patients with endometriosis and 50 normal endometrial tissues were collected from January 2019 to December 2022.Twelve healthy female C57BL/6 mice were selected to establish a mouse endometriosis model by autologous peritoneal wall transplantation of uterine tissue,and the mice were randomly divided into SHP2-NC group(n=6)and SHP2-shRNA group(n=6).RAW264.7 cells with stable SHP2 gene knockdown were constructed by lentivirus infection.It was injected through a mouse tail vein.HE staining,Western blot,TUNEL staining,CCK-8 and tube formation tests were used to detect the patho-logical changes of endometrial tissue,CD68+macrophage-related protein expression,endometrial cell apoptosis,endometrial cell proliferation and endothelial cell angiogenesis,respectively.Results Compared with normal endometriosis tissues,the expression level of SHP2 in CD68+macrophages in endometriosis tissues was increased(P＜0.05).In the SHP2-shRNA group,mesenchymal cells appeared in endometrial abnormal lesions,but glandular and vascular formation decreased.Mean-while,the expression levels of SHP2,NADPH oxidase-2(NOX2),NADPH oxidase-4(NOX4),cyclooxygenase-2(COX2),vascular endothelial growth factor(VEGF)and Bcl-2 were lower than those of SHP2-NC group,while the protein expression levels of p53,Caspase-3 and Bax were higher than those of SHP2-NC group(P＜0.05).Compared with the SHP2-NC group,the number of apoptosis of intima cells in the SHP2-shRNA group was increased,the cell proliferation activity was decreased,and the number of endothelial cell angiogenesis was decreased(P＜0.05).The protein expression levels of p-PI3K,NOX4,COX2 and VEGF after SHP2-shRNA intervention were decreased compared with SHP2-NC intervention(P＜0.05),while the expression levels of PTEN and p53 were increased(P＜0.05).After LY294002 intervention,the expression levels of p-PI3K,NOX4,COX2 and VEGF were decreased(P＜0.05),while the expression levels of PTEN and p53 were increased,as compared with those before intervention(P＜0.05).However,after 740 Y-P intervention,the expression levels of p-PI3K,NOX4,COX2 and VEGF were increased(P＜0.05),while the expression levels of PTEN and p53 were decreased,as compared with those before intervention(P＜0.05).Conclusion SHP2 enhances oxidative stress and endothelial angio-genesis by promoting PI3K/PTEN pathway activity,while inhibiting endometrial cell apoptosis.