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目的检测全反式维A酸(all-trans-retinoic acid,ATRA)和佛波酯(PMA)对人子宫内膜基质细胞(HESC)分泌血管内皮生长因子(VEGF)的增强作用。方法通过ELISA、real time PCR检测ATRA、PMA对在VEGF m RNA和蛋白水平上的影响;通过Western-blotting检测ATRA、PMA对HESC表达VEGF及其受体的影响;观察ATRA对VEGF分泌的剂量和时间依赖性;评价ATRA对VEGF的特异性;使用H_2O_2代替ATRA联合PMA刺激HESC,探讨ATRA介导VEGF分泌增加的可能性途径。结果单独使用ATRA不能改变VEGF的mRNA和蛋白表达,单独使用PMA虽然能够增加VEGF的m RNA表达但并不能增加VEGF蛋白的表达,当共同使用ATRA和PMA时却明显地提高VEGF蛋白的表达量。ATRA对VEGF分泌的促进作用具有明显的剂量和时间依赖性,且该作用是特异性。H2O2可以模拟ATRA增加PMA对HESC分泌VEGF的刺激作用。结论ATRA可以联合PMA迅速增强HESC的VEGF表达,这种作用可能是通过氧化还原反应介导的。
Objective To investigate the potentiation effect of all-trans-retinoic acid (ATRA) and phorbol myristate (PMA) on the secretion of vascular endothelial growth factor (VEGF) from human endometrial stromal cells (HESC). Methods The effect of ATRA and PMA on the expression of VEGF m RNA and protein was detected by ELISA and real time PCR. The effect of ATRA and PMA on the expression of VEGF and its receptor in HESC was detected by Western-blotting. The effect of ATRA on VEGF secretion and Time dependence; evaluate the specificity of ATRA for VEGF; use H_2O_2 instead of ATRA combined with PMA to stimulate HESC to explore the possibility of ATRA-mediated increase in VEGF secretion. Results ATRA alone could not change the mRNA and protein expression of VEGF. Although PMA alone could increase the mRNA expression of VEGF but did not increase the expression of VEGF, the expression of VEGF protein was significantly increased when ATRA and PMA were used together. The promoting effect of ATRA on VEGF secretion has obvious dose and time dependence, and the effect is specific. H2O2 can simulate ATRA increase PMA stimulation of HESC secretion of VEGF. Conclusion ATRA can rapidly enhance VEGF expression of HESC in combination with PMA, and this effect may be mediated by redox reaction.