Dehghan Maryam, Shahbazi Shirin, Salehnia Mojdeh
Anatomy Department, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Medical Genetic Department, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
J Family Reprod Health. 2021 Jun;15(2):91-98. doi: 10.18502/jfrh.v15i2.6449.
The aim of this study was to evaluate the effect of lysophosphatidic acid (LPA) supplementation during in vitro culture and transplantation of mouse ovaries on the follicular development and expression of vascular endothelial growth factor (VEGF) as an angiogenesis factor at the mRNA and protein levels. Three weeks old mice ovaries were cultured in the presence and absence of LPA for 24 hours, then they were capsulated in sodium alginate in the presence and absence of LPA as four experimental groups. After transplantation the vaginal smears were performed daily to evaluate the initiation of the estrous cycle. The morphology and follicular distribution were analyzed at the first and fourth estrous cycles using hematoxylin and eosin staining. Then in the groups that showed higher and lower follicular development the immunohistochemistry assay was conducted to identify VEGF protein expression, and the real time RT-PCR was done to analyze the expression of gene at the first estrus cycle. The large size follicles and also the corpus luteum were prominent in all transplanted groups at fourth estrus cycle in comparison with intact control groups. The statistically lowest percentage of small size follicles and the highest percentages of large size follicles were seen in LPA/LPA group (p<0.05). The expression ratio of to was significantly higher in this group in comparison with non-LPA treated and intact control groups (p <0.05). LPA as an angiogenesis factor increases the follicular development in transplanted ovaries but it causes early discharge of ovarian reserve.
本研究的目的是评估在小鼠卵巢体外培养和移植过程中补充溶血磷脂酸（LPA）对卵泡发育以及血管内皮生长因子（VEGF，一种血管生成因子）在mRNA和蛋白质水平表达的影响。将三周龄小鼠的卵巢在有和无LPA的情况下培养24小时，然后在有和无LPA的情况下用海藻酸钠包裹，分为四个实验组。移植后每天进行阴道涂片以评估发情周期的开始。在第一个和第四个发情周期使用苏木精和伊红染色分析形态和卵泡分布。然后在卵泡发育较高和较低的组中进行免疫组织化学分析以鉴定VEGF蛋白表达，并在第一个发情周期进行实时RT-PCR以分析基因表达。与完整对照组相比，在第四个发情周期时，所有移植组中的大卵泡以及黄体都很明显。在LPA/LPA组中，小卵泡的统计学最低百分比和大卵泡的最高百分比被观察到（p<0.05）。与未用LPA处理的组和完整对照组相比，该组中VEGF的表达率显著更高（p<0.05）。LPA作为一种血管生成因子可增加移植卵巢中的卵泡发育，但会导致卵巢储备的过早排出。
