Xu Shaoyuan, Li Jie, Chen Xiaoyan, Liu Beiyu
Reproductive Medicine Center, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, China.
Human Reproductive Center of the First Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510080, China.
Biochem Res Int. 2020 Oct 23;2020:8892930. doi: 10.1155/2020/8892930. eCollection 2020.
Whether changes in vascular endothelial growth factor (VEGF) and annexin IV during implantation are regulated through the LH/hCG-R needs further research. To investigate the mechanism of hCG on the expression of annexin IV and VEGF in human endometrial cells.
Endometrial cells were isolated and identified from human specimens. The proportion of glandular and epithelial cells was analyzed. Annexin IV and VEGF were analyzed by qRT-PCR (mRNA), western blot (proteins), and immunohistochemistry (proteins). Protein location was identified by immunohistochemistry. The cells were cultured with hCG, hCG/PD98059 (a MAPK inhibitor), or no treatment (control).
The proportions between the glandular epithelial cells and stromal cells at inoculation and when adding hCG were 25.8 ± 0.2% and 27.8 ± 0.04%, respectively ( > 0.05). LH/hCG-R, annexin IV, and VEGF were found in the cytoplasm of endometrial cells. After 2, 6, 12, and 24 h of hCG treatment, compared with 1 h, VEGF mRNA was increased by 1.25-fold, 3.19-fold, 4.21-fold, and 4.86-fold and annexin IV by 2.23-fold, 3.37-fold, 5.14-fold, and 5.02-fold. Compared with the control group, annexin IV mRNA and protein were increased in the hCG and hCG/PD98059 groups (mRNA/protein: 1.99-fold/1.80-fold and 2.33-fold/1.93-fold, < 0.05). Compared with the control group, VEGF mRNA and protein were increased in the hCG group (mRNA/protein: 2.30-fold/1.86-fold), but not in the hCG/PD98059 group.
hCG could upregulate the mRNA and protein expression of annexin IV and VEGF. The upregulation of annexin IV by hCG could not be inhibited by PD98059, but the upregulation of VEGF by hCG could.
着床期间血管内皮生长因子（VEGF）和膜联蛋白IV的变化是否通过促黄体生成素/人绒毛膜促性腺激素受体（LH/hCG-R）调控尚需进一步研究。本研究旨在探讨人绒毛膜促性腺激素（hCG）对人子宫内膜细胞中膜联蛋白IV和VEGF表达的影响机制。
从人标本中分离并鉴定子宫内膜细胞，分析腺上皮细胞和基质细胞的比例。采用实时定量逆转录聚合酶链反应（qRT-PCR，检测mRNA）、蛋白质免疫印迹法（western blot，检测蛋白质）和免疫组织化学法（检测蛋白质）分析膜联蛋白IV和VEGF。通过免疫组织化学鉴定蛋白质定位。将细胞分别用hCG、hCG/PD98059（一种丝裂原活化蛋白激酶抑制剂）处理或不处理（作为对照）。
接种时及添加hCG时，腺上皮细胞与基质细胞的比例分别为25.8±0.2%和27.8±0.04%（P>0.05）。在子宫内膜细胞的细胞质中发现了LH/hCG-R、膜联蛋白IV和VEGF。hCG处理2、6、12和24小时后，与处理1小时相比，VEGF mRNA分别增加了1.25倍、3.19倍、4.21倍和4.86倍，膜联蛋白IV分别增加了2.23倍、3.37倍、5.14倍和5.02倍。与对照组相比，hCG组和hCG/PD98059组的膜联蛋白IV mRNA和蛋白质均增加（mRNA/蛋白质：1.99倍/1.80倍和2.33倍/1.93倍，P<0.05）。与对照组相比，hCG组的VEGF mRNA和蛋白质增加（mRNA/蛋白质：2.30倍/1.86倍），而hCG/PD98059组则未增加。
hCG可上调膜联蛋白IV和VEGF的mRNA和蛋白质表达。hCG对膜联蛋白IV的上调作用不能被PD98059抑制，但对VEGF的上调作用可以被抑制。
