番茄红素对小鼠原始卵泡发育的影响及机制
Effect and mechanism of lycopene on primordial follicle development in mice
杨雪 1唐紫雯 1刘冬菊 1彭毛卓玛 1莫罗钰 1马骏 1黄瑞雪 1殷实2
作者信息
- 1. 西南民族大学畜牧兽医学院,四川成都 610041
- 2. 西南民族大学畜牧兽医学院,四川成都 610041;西南民族大学现代生物技术国家民委重点实验室,四川成都 610041
- 折叠
摘要
旨在探究体外培养条件下番茄红素对小鼠原始卵泡激活的作用及机制.分离出生后3 d的ICR小鼠卵巢并对其在体外连续培养4 d,在培养过程中添加不同浓度番茄红素0(对照组)、5、50和500 μmol/L,通过HE染色分析不同浓度番茄红素对小鼠原始卵泡发育的影响,通过过碘酸-希夫(periodic acid Schiff,PAS)染色及TUNEL染色检测卵泡闭锁及细胞凋亡情况,通过增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)染色检测颗粒细胞增殖情况,应用ELISA检测卵巢中活性氧(reactive oxygen species,ROS)水平,通过Real-time PCR检测部分凋亡及抗氧化基因表达,并通过 Western blot检测卵泡激活关键信号通路PI3K-AKT中部分成员蛋白的表达.结果显示,与对照组相比,500 μmol/L番茄红素浓度培养条件下原始、过渡态、初级及次级卵泡数量均显著升高(P＜0.05),而退化卵泡及凋亡细胞数量显著减少(P＜0.05),处于增殖状态的颗粒细胞数量显著增多(P＜0.05);与对照组相比,500μmol/L处理组中ROS水平明显降低(P＜0.05),部分抗凋亡基因及抗氧化基因,包括Bcl-2、Xiap、Apaf-1、Cat、Prdx1、Gpx1的表达显著升高(P＜0.05),而促凋亡基因Bax、Caspase-3、Caspase-8的表达显著下调(P＜0.05);500μmol/L处理组中PTEN、AKT和FOXO3A蛋白的表达水平与对照相比明显下调(P＜0.05),而磷酸化AKT及磷酸化FOXO3A蛋白的表达水平显著上调(P＜0.05).综上所述,500 μmol/L番茄红素可能通过增强细胞的抗氧化性和调控PI3K-AKT通路部分蛋白的表达促进原始卵泡的存活和激活.本试验为深入探讨番茄红素调控原始卵泡发育的分子机制提供了一定的试验数据,并为原始卵泡发育障碍引发的相关不孕症的治疗提供了新的思路.
Abstract
The aim of this study was to investigate the effect and mechanism of lycopene on the ac-tivation of primordial follicles in mice cultured in vitro.The ovaries of ICR mice at 3 d after birth were isolated and cultured for 4 d in vitro,during which 0(control group),5,50 and 500 μmol/L lycopene were added.The effects of lycopene on primordial follicle development in mice were ana-lyzed by HE staining.Periodic acid Schiff(PAS)and TUNEL staining were used to detect follicu-lar atresia and cell apoptosis.Proliferating cell nuclear antigen(PCNA)staining was performed to detect the proliferation of granulosa cells.The level of reactive oxygen species(ROS)in ovary was detected by ELISA,and the expressions of some apoptosis and antioxidant-related genes were de-tected by real-time PCR.Western blot was used to detect the protein expressions of some members in PI3K-AKT pathway,a key signaling pathway for follicle activation.The results showed that the number of primordial,transitional,primary and secondary follicles increased significantly(P＜0.05)under 500 μmol/L lycopene concentration culture compared with the control group,and the number of degenerated follicles and apoptotic cells in 500 μmol/L treatment group were signifi-cantly reduced(P＜0.05),while the number of proliferating granulosa cells was significantly in-creased(P＜0.05).Compared with control group,the level of ROS in 500 μmol/L lycopene treat-ment group decreased significantly(P＜0.05),and the expressions of some anti-apoptotic genes and antioxidant genes,including Bcl-2,Xiap Apaf-1,Cat,Prdx1 and Gpx1,were significantly in-creased(P＜0.05)in 500 μmol/L treatment group,while the expressions of pro-apoptotic genes Bax,Caspase 3 and Caspase 8 were significantly down-regulated(P＜0.05).The protein levels of PTEN,AKT and FOXO3A in 500 μmol/L group were significantly down-regulated(P＜0.05)compared with the control group,while the levels of phosphorylated AKT and phosphorylated FOXO3A were significantly up-regulated(P＜0.05).In conclusion,the concentration of 500 μmol/L lycopene may promote the survival and activation of primal follicles by enhancing the antioxidant activity of cells and regulating the expression of some proteins in PI3K-AKT pathway.This study provided some experimental data for further investigation of the molecular mechanism of lycopene regulating primordial follicle development,and provided a new idea for the treatment of infertility caused by primordial follicle development disorder.
关键词
番茄红素/原始卵泡/卵泡激活/抗氧化/PI3K-AKT信号通路Key words
lycopene/primordial follicle/follicular activation/antioxidant/PI3K-AKT signaling path-way引用本文复制引用
基金项目
四川省自然科学基金(2022NSFSC1709)
西南民族大学"双一流"建设项目(XM2023008)
出版年
2024
NETL
NSTL
万方数据