Ha L-X, Wu Y-Y, Yin T, Yuan Y-Y, Du Y-D
Center for Reproductive Medicine, General Hospital, Ningxia Medical University, Yinchuan, Ningxia, China.
Key Laboratory of Fertility Preservation and Maintenance, Ministry of Education, Ministry of Education, Ningxia Medical University, Yinchuan, Ningxia, China.
J Physiol Pharmacol. 2021 Dec;72(6). doi: 10.26402/jpp.2021.6.13. Epub 2022 Apr 24.
The present study aimed to investigate the possible mechanism of action of tumor necrosis factor-alpha (TNF-α) in endometrial glucose transporter-4 (GLUT-4) expression regulation by detecting the expression levels of endometrial inflammatory factors and GLUT-4 in patients with polycystic ovary syndrome (PCOS). A total of 140 patients were included in this study and divided into four groups: the PCOS group, the obesity + PCOS group, the normal group, and the obesity group (n = 35 each). The general clinical data of all patients were collected, and their expression levels of TNF-α, nuclear factor kappa B p65 (NF-κBp65), and GLUT-4 in the endometrium were tested via immunohistochemistry. Endometrial stromal cells were cultured in vitro and treated with TNF-α or pyrrolidine dithiocarbamate (PDTC) + TNF-α, and the expression levels of NF-κBp65, phospho-NF-κBp65 (p-NF-κBp65), and GLUT-4 were tested using Western blotting (WB) before and after treatment. As a result, we got: 1) Compared with the normal group, the abundance of the protein for TNF-α and NF-κBp65 in the endometrium of the patients with PCOS was elevated, while the expression level of GLUT-4 was decreased; the difference was statistically different (P < 0.05). The comparison between the obesity + PCOS group and the PCOS group yielded the same results. 2) According to the WB results, compared with the normal group, the abundance of the protein for endometrial GLUT-4 was decreased in the PCOS group, and the expression levels of p-NF-κBp65 and NF-κBp65 were increased in the obesity + PCOS group; the differences were statistically different (P < 0.05). The addition of TNF-α could decrease the abundance of the protein for GLUT-4 and increase the abundance of the protein for p-NF-κBp65. After treatment with PDTC + TNF-α, the abundance of the protein for p-NF-κBp65 decreased and GLUT-4 increased compared with the TNF-α group; these values were close to those of the control group. We concluded that the abundance of the proteins for local inflammatory factors in the endometrial cells of patients with PCOS was increased, indicating that TNF-α could affect the expression of endometrial GLUT-4 in such patients by activating the p-NF-κBp65 signaling pathway.
本研究旨在通过检测多囊卵巢综合征（PCOS）患者子宫内膜炎症因子和葡萄糖转运蛋白4（GLUT-4）的表达水平，探讨肿瘤坏死因子-α（TNF-α）在子宫内膜GLUT-4表达调控中的可能作用机制。本研究共纳入140例患者，分为四组：PCOS组、肥胖+PCOS组、正常组和肥胖组（每组n = 35）。收集所有患者的一般临床资料，采用免疫组织化学法检测其子宫内膜中TNF-α、核因子κB p65（NF-κBp65）和GLUT-4的表达水平。体外培养子宫内膜基质细胞，用TNF-α或吡咯烷二硫代氨基甲酸盐（PDTC）+TNF-α处理，处理前后采用蛋白质印迹法（WB）检测NF-κBp65、磷酸化NF-κBp65（p-NF-κBp65）和GLUT-4的表达水平。结果如下：1）与正常组相比，PCOS患者子宫内膜中TNF-α和NF-κBp65蛋白丰度升高，而GLUT-4表达水平降低；差异有统计学意义（P < 0.05）。肥胖+PCOS组与PCOS组比较结果相同。2）根据WB结果，与正常组相比，PCOS组子宫内膜GLUT-4蛋白丰度降低，肥胖+PCOS组p-NF-κBp65和NF-κBp65表达水平升高；差异有统计学意义（P < 0.05）。添加TNF-α可降低GLUT-4蛋白丰度，增加p-NF-κBp65蛋白丰度。与TNF-α组相比，PDTC+TNF-α处理后，p-NF-κBp65蛋白丰度降低，GLUT-4升高；这些值接近对照组。我们得出结论，PCOS患者子宫内膜细胞中局部炎症因子蛋白丰度增加，表明TNF-α可能通过激活p-NF-κBp65信号通路影响此类患者子宫内膜GLUT-4的表达。
