小檗碱对猪卵巢卵母细胞体外成熟及其脂滴含量和分布的影响

摘要

[目的]探明不同形态的猪卵巢中卵母细胞体外成熟效果与脂滴的相关性,以及小檗碱对其的影响.[方法]将卵泡型卵巢GV期和黄体型卵巢GV期卵母细胞分别用添加小檗碱的成熟液体外成熟,观察体外成熟效果;用尼罗红染色的方法测定成熟前后的卵母细胞的脂滴含量,并观察成熟后各组卵母细胞脂滴的分布.[结果]卵泡型卵母细胞的颗粒细胞扩散率和第一极体排出率均显著高于黄体型卵母细胞(P＜0.05),小檗碱(Berberine,Ber)组卵母细胞的颗粒细胞扩散率显著高于对照组(P＜0.05),第一极体排出率极显著高于对照组(P＜0.01);体外成熟后,对照组卵母细胞中脂滴含量显著降低(P＜0.05),小檗碱组极显著降低(P＜0.01),且极显著低于对照组(P＜0.01).黄体型卵母细胞的脂滴含量则显著低于卵泡型(P＜0.05).猪卵母细胞成熟后的脂滴分布为分散型、外围型和不规则型,卵泡型卵母细胞脂滴主要呈分散分布,黄体型卵母细胞脂滴主要呈外围分布,并且黄体型卵母细胞脂滴呈不规则分布多于卵泡型.[结论]小檗碱能明显促进猪卵母细胞体外成熟,且卵泡型成熟效果显著好于黄体型;体外成熟后,卵母细胞脂滴逐渐减少,小檗碱组脂滴含量极显著减少,且黄体型卵母细胞脂滴含量明显减少;猪卵母细胞内脂滴的分布与卵巢所处的时期有关.%[Objective]The aims of the paper are to investigate the correlation between in vitro maturation(IVM) and lipid droplets of oocytes from different forms of porcine ovarian and the effect of berberine(Ber).[Methods] Porcine oocytes of GV phase from follicular ovarian and luteal ovarian were matured in IVM medium with Berberine,to observe the effect of IVM;The lipid droplet content of oocytes were measured by the method of Nile red before and after IVM and the distribution of lipid droplet of oocytes of each group after IVM were observed.[Results] Comparing with oocytes from luteal ovarian,granular cells proliferation rate and the polar body I(pbI) formation rate from follicular ovarian oocytes were significantly higher(P＜0.05),granular cells proliferation rate and pbI formation rate of oocytes in berberine group were significantly higher than the control group (P ＜0.05,P＜0.01 respectively).After IVM,the lipid droplets content of oocytes of the control group were significantly decreased (P＜0.05).That of berberine group was highly significantly decreased(P ＜0.01),and highly significantly lower than the control group (P＜0.01).The lipid droplet content of oocytes from luteal ovarian was significantly lower than that of the follicular ovarian(P＜0.05).After the IVM,the distribution of lipid droplets in the oocytes were divided into three types:the scattered type,peripheral type and irregular type.The lipid droplets of follicular ovarian oocytes were mainly scattered distribution,the luteal ovarian oocytes were mainly peripheral distribution,the irregular distribution of the lipid droplets in luteal ovarian oocytes were more than that of the follicular ovarian oocytes.[Conclusion] Berberine can significantly promote the in vitro maturation of porcine oocytes,and the maturation of follicular ovarian oocytes was significantly better than that of luteal ovarian oocytes.After IVM,the lipid droplets content of oocytes decreased gradually in the control group,and significantly decreased in the berberine group.The lipid droplets content of luteal ovarian oocytes was significantly decreased.The distribution of lipid droplets in porcine oocytes was related to the period of ovary.