解放军医学杂志2024，Vol.49Issue(6)：679-685,7.DOI:10.11855/j.issn.0577-7402.2633.2024.0229
VEGF对小鼠卵巢类固醇合成相关基因表达的影响及其机制
Effect of VEGF on the expression of genes related to ovarian steroid synthesis in mice and its mechanism
摘要
Abstract
Objective To investigate the effect of vascular endothelial growth factor(VEGF)on the expression of genes related to ovarian steroid synthesis in mice and its underlying mechanism.Methods A transgenic mouse model with tetracycline-reversible regulation of VEGF expression was used,and the genotype of mice was identified by polymerase chain reaction(PCR).Twenty mice were divided into normal VEGF expression group(Dox+,n=10)and VEGF expression inhibition group(Dox-,n=10)by feeding them doxycycline.Western blotting was used to detect the expression of VEGF protein in ovarian tissues.Fluorescence quantitative PCR was used to detect the mRNA expression of VEGF,KDR and genes known to play roles in follicle development,such as follicle-stimulating hormone(FSH)and inhibin B(INHBB).HE staining was used to observe changes in ovarian tissue.Total RNA was extracted from mouse ovarian tissues for transcriptome sequencing,and the relevant differential genes were analyzed by FPKM and log2FC values.Results Compared with the Dox+group,the mRNA and protein levels of VEGF in the Dox-group significantly reduced,and the mRNA levels of KDR also significantly decreased(P<0.05).HE staining results showed that compared with the Dox+group,follicular development was impaired and atresia follicles appeared in the Dox-group.Sequencing analysis identified that significant differences in follicular development-related genes and steroid synthesis-related genes between the two groups(P<0.05).Enrichment analysis showed that VEGF in mouse ovaries mainly regulates ovarian steroidogenesis and other pathways.Fluorescence quantitative PCR results demonstrated that compared with the Dox+group,the follicular development-related genes(INHBB and FSHR)in the ovarian tissues of the Dox-group were significantly up-regulated(P<0.05),whereas the key genes of steroid synthesis(StAR,CYP11A1,3β-HSD)were significantly down-regulated(P<0.05).The quantitative results were basically consistent with the sequencing results.Conclusion Mice with inhibited VEGF exhibited ovarian follicular dysplasia,potentially due to the mechanism whereby VEGF inhibition downregulated the expression of genes associated with steroid synthesis,such as FSH and INHBB,thereby obstructing cholesterol metabolism.关键词
血管内皮生长因子/卵泡发育/性类固醇激素/转录组测序Key words
vascular endothelial growth factor/follicular development/sexual steroid hormones/transcriptome sequencing分类
医药卫生引用本文复制引用
张智慧,高鸿霞,王国庆,侯巍,邹畅,芦小单..VEGF对小鼠卵巢类固醇合成相关基因表达的影响及其机制[J].解放军医学杂志,2024,49(6):679-685,7.基金项目
This work was supported by the Natural Science Foundation of Jilin Province(20240101007JC) （20240101007JC）
the Natural Science Foundation of Jilin Province(YDZJ202201ZYTS148),and the Jilin Provincial Science and Technology Department Innovation Team Project(20220508080RC) 吉林省自然科学基金(20240101007JC) （YDZJ202201ZYTS148）
吉林省自然科学基金项目(YDZJ202201ZYTS148) （YDZJ202201ZYTS148）
吉林省科技厅创新团队项目(20220508080RC) （20220508080RC）
