为探讨性腺激素对山羊子宫内膜细胞分泌活性的影响,基于套皿培养技术构建永生化山羊子宫内膜上皮细胞(EEC)与基质细胞(ESC)共培养体外研究模型,通过ELISA检测性腺激素E2和/或P4对单独培养EEC中TNF-α与TGF-β分泌活性的调节作用以及ESC对该培养体系的影响。结果显示:与未加激素对照组相比,单独或混合添加E2和P4均可显著增加单独培养EEC向顶端分泌TNF-α水平(P〈0.05),而对细胞基底端TNF-α分泌活性影响不显著;E2单独作用较对照组显著降低EEC向细胞顶端分泌TGF-β水平(P〈0.05),P4单独或与E2共同作用下则主要对单独培养EEC顶端及基底端分泌TGF-β水平具有显著增强作用(P〈0.05)。对于EEC-ESC共培养细胞模式,E2和/或P4显著降低EEC顶端和基底端TNF-α分泌水平(P〈0.05);EEC顶端TGF-β分泌水平在E2和/或P4作用下显著降低(P〈0.05),而EEC基底端TGF-β分泌水平不受激素作用所影响。激素对单独培养及与ESC共培养模式下的极化EEC顶端及基底端TNF-α与TGF-β分泌水平的差异,提示ESC对激素作用下山羊子宫内膜上皮细胞分泌方向及分泌水平的重要调节作用。
The aim of this study was to study the effect of sex gland hormones on the secretory activity to endometrial cells of goats. Based on the hanging cell coculture insert system, immor- talization of goats endometrial epithelial cells (EEC) and stromal cells (ESC) has trained cocul- ture in vitro studies model, the different levels and combinations of progesterone (P4) and estra- diol (E2) were added in medium, without or with ESC together, TNF-~ and TGF β secretory ac tivity of EEC were determined by ELISA. The results showed that compared with no hormone control group, the secretion of TNF-a on EEC apical was enhanced markedly by P4 (P~0.05) when added alone or together E2 and P4, but not significant on basolateral; TGF t3 secretory ac- tivity of both EEC apical and basolateral were reduced markedly by different concentrations and combinations of E2 and/or P4 (P~0.05), but TGF-β on EEC apical was restrained significantly by E2alone (P〈0.05). About coculture with ESC together, both apical and basolateral of TNF a secretion were repressed significantly by sex hormones E2/P4 (P〈0.05) ; TGF-β on EEC apicalwas restrained significantly by E2/P4 (P〈0.05), but not significant on basolateral. Our results indicated that the sex hormones in coculture mode alone or together with ESC, all can regulate the secretion of TNF-a and TGF-β with different levels, suggested that ESC play an important regulatory role to the secrete direction and secretion levels of goat EEC when coculture with sex hormones.