晚期糖基化终末产物对内皮细胞前列腺素合成的影响及可能机制

Effects of Advanced Glycation End Products on Prostacyclin and Prostaglandin E_2 Expression in Culture Endothelial Cell and Its Mechanism

目的探讨晚期糖基化终末产物对内皮细胞前列环素、前列腺素E2合成的影响及可能机制。方法不同浓度的晚期糖基化终末产物作用内皮细胞,酶联免疫吸附法测定前列环素的稳定代谢产物6-酮—前列腺素F1α和前列腺素E2的表达。逆转录聚合酶链反应、免疫细胞化学测定环氧合酶2 mRNA和蛋白表达的改变。晚期糖基化终末产物受体、核因子κB单/双基因反义RNA对晚期糖基化终末产物刺激内皮细胞分泌6-酮—前列腺素F1α、前列腺素E2的影响。结果晚期糖基化终末产物引起内皮细胞分泌6-酮—前列腺素F1α、前列腺素E2增加(P<0.01),具有剂量依赖关系。晚期糖基化终末产物引起内皮细胞环氧合酶2的mRNA和蛋白表达增加(P<0.01)。晚期糖基化终末产物受体、核因子κB单/双基因反义RNA可减轻晚期糖基化终末产物刺激的内皮细胞分泌6-酮—前列腺素F1α、前列腺素E2(P<0.05或0.01),双基因的抑制效果更明显(P<0.05)。结论晚期糖基化终末产物可能通过晚期糖基化终末产物受体、核因子κB途径使内皮细胞的环氧合酶2表达增加,从而引起前列环素、前列腺素E2合成增加,这个机制可能参与了晚期糖基化终末产物所致的血管炎症损伤。

Aim To detect the effects of advanced glycation end products （AGE） on prostacyclin and prostaglandin E2 expressions in culture endothelial cell and its possible mechanism. Methods ECV304 cell were exposed to 50, 100, 200, 400 mg/L AGE or human serttm albumin for 24 hours. The levels of 6-keto-prostaglandin F1a. （ PGF1a ） and prostaglandin （ PGE2 ） in the supernatants were measured by enzyme linked immtmosorbent assay （ ELISA）. The expressions of cyclooxyenase- 2 （COX-2） mRNA and protein were detected by reverse transcription polymerase chain reaction （RT-PCR） and iwantmohistochemistric assay. The effects of receptor for AGE, nuclear factor-kappaB （NF-κB） single/double gene antisense RNA on the productions of 6-keto- PGF1a and PGE2 induced by AGE were measured by ELISA. Results The productions of 6-keto- PGF1a and PGE2 in the supernatants were higher in the cell treated by AGE, with a dose-dependent manner （ P 〈 0.01 ）. The levels of COX-2 mRNA and protein were significantly higher in the cell treated by AGE （ P 〈 0.01 ）. Receptor for AGE, NF-κB single/ double gene antisense RNA could inhibit the productions of 6-keto-PGF1a and PGE2 treated by AGE （ P 〈 0.05 or 0.01 ）. The inhibition of double gene antisense RNA were more effective than single gene antisense RNA （ P 〈 0.05 ）. Conclusion AGE, via receptor for AGE, NF-κB signaling, induced upregulation of COX-2, resulting in induced prostacyclin and PGEa synthesis. This chain of events might contribute to the pathogenesis of inflammatory vascular injury.