MIF在不同直径猪卵泡的表达变化规律

为分析巨噬细胞迁移抑制因子(Macrophage migration inhibitory factor,MIF)在不同直径健康猪卵泡上的表达变化规律,将收集到的90个健康母猪卵巢,通过ELISA检测不同直径卵泡液中MIF质量浓度,通过实时荧光定量PCR和Western blot检测颗粒细胞中MIF基因mRNA和蛋白的表达水平,采用免疫荧光检测MIF在颗粒细胞中的分布情况。结果显示:1)中等直径卵泡(3～4mm)液中MIF的质量浓度(54.17±7.86ng/mL)显著高于大卵泡(6mm,32.29±1.47ng/mL)和小卵泡(1～2mm,38.89±0.98ng/mL)(P0.05);2)中等直径卵泡颗粒细胞中MIF基因mRNA和蛋白质表达量也显著高于大卵泡和小卵泡的表达量(P0.05);3)MIF也在不同直径卵泡颗粒细胞中的细胞核和细胞质表达,但多集中于细胞质中。综上,随着猪卵泡发育过程,MIF在中等直径卵泡中表达量最高。本研究为母猪卵泡发育和排卵提供新的线索。

Expression of MIF in porcine follicles of different diameters

In order to analyze the expression of macrophage migration inhibitory factor(MIF)in different diameters follicles of healthy pigs, 90 ovaries of healthy sows were collected. The concentration of MIF in follicular fluid of different diameters were detected by ELISA. The expression of MIF mRNA and protein in granulosa cells were detected by quantitative real-time PCR and Western blot, and the distribution of MIF in granulosa cells was assayed by immunofluorescence. The results showed that: 1)The concentration in medium follicles(diameter 3-4 mm)fluid(54. 17±7. 86 ng/mL)was significantly higher than that in large follicles(diameter>6 mm, 32. 29±1. 47 ng/mL)and small follicles(diameter 1-2 mm, 38. 89±0. 98 ng/mL)(P<0. 05); 2)The expression of MIF mRNA and protein in granulosa cells of medium diameter follicles was also significantly higher than that in large follicles and small follicles(P<0. 05); 3)MIF was also expressed in the nucleus and cytoplasm of granulosa cells with different diameters, but mostly concentrated in the cytoplasm. In summary, with the development of porcine follicles, the expression of MIF was the highest in medium-sized follicles(diameter 3-4 mm). This study provides new clues for follicular development and ovulation.