Inhibitory effect of retinoic acid on the development of immature porcine granulosa cells to mature cells
Inhibitory effect of retinoic acid on the development of immature porcine granulosa cells to mature cells
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DOI：
10.1677/jme.0.0250053
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发表时间：
2000-08-01
影响因子：
3.5
通讯作者：
Fujihara, N
中科院分区：
文献类型：
--
作者：
Hattori, MA;Takesue, K;Fujihara, N
研究了维甲酸（RA）对猪卵巢颗粒细胞分化的影响。将颗粒细胞预培养15 h，然后用FSH培养48 h，再用LH处理24 h，以诱导其转化为黄体细胞。在细胞暴露于1 μ M类维生素A（RA，视黄醇和视黄醇）87小时后，通过使用半定量RT-PCR进行LH受体mRNA表达（颗粒细胞分化的指标）的分析。结果表明，LH受体mRNA水平下降，RA对这些水平的影响比其他两种维甲酸更强。当细胞在未成熟阶段（加入FSH之前）或发育早期（加入FSH后0-24 h）暴露于RA时，LH受体mRNA的表达大大减少。未成熟细胞用RA培养15 h，再用FSH洗涤培养48 h，用LH洗涤培养24 h，LH受体mRNA的表达无逆转。然而，在分化的细胞中（加入FSH后24小时），RA不再有任何抑制作用。当未成熟细胞暴露于IA时，FSH诱导的c-fos mRNA表达显著降低。相反，c-jun和转录激活因子-4 mRNA的表达保持不变。而forskolin对c-fos mRNA的表达无明显影响。结果表明，RA是一个有效的抑制剂在未成熟阶段的猪颗粒细胞分化，可能通过减少FSH受体的表达，但RA不抑制分化的成熟阶段的细胞。
The present study investigated the effect of retinoic acid (RA) on the differentiation of granulosa cells prepared from porcine ovaries. The granulosa cells were precultured for 15 h, then cultured for 48 h with FSH and further treated for 24 h with LH in order to induce their transformation into luteal cells. After the cells had been exposed to 1 mu M retinoids (RA, retinal and retinol) for 87 h, analysis of the LH receptor mRNA expression, an indicator of granulosa cell differentiation, was carried out by using semiquantitative RT-PCR. The results showed that there was a decrease in LH receptor mRNA levels, and that RA had a more potent effect on these levels than the other two retinoids. When cells were exposed to RA in the immature stage (before the addition of FSH) or the early stage of development (0-24 h after the addition of FSH), expression of LH receptor mRNA was greatly diminished. When the immature cells were cultured for 15 h with RA, then washed and cultured for 48 h with FSH and for 24 h with LH, the expression of LH receptor mRNA was not reversed. In the differentiated cells (24 h after the addition of FSH), however, RA no longer had any inhibitory effect. When the immature cells were exposed to IA, FSH-induced expression of c-fos mRNA was markedly decreased. In contrast, expression of c-jun and activating transcription factor-4 mRNAs remained constant. However, the expression of c-fos mRNA was not decreased by forskolin. The results indicate that RA is a potent inhibitor in the immature stage of porcine granulosa cell differentiation, probably through decreased expression of FSH receptor, but that RA does not inhibit differentiation in the mature stage of the cells.