内皮素-1对小鼠脑微血管内皮细胞环氧化酶-2、前列腺素E2表达的影响

Effect of endothelin-1 on mice brain microvascular endothelial cells cyclooxygenase-2 and prostaglandin E2 expresstion

目的：探讨内皮素-1（ET-1）对小鼠脑微血管内皮细胞中环氧化酶-2（COX-2）及前列腺素E2（PGE2）表达的影响。方法：培养小鼠脑微血管内皮细胞bEnd.3株,分别加入不同浓度（1、10、100和1 000 nmol/L）的ET-1处理2、4、6、16和24 h,用Western blot技术检测不同浓度ET-1处理不同时间后细胞内COX-2表达的变化;另取生长良好的细胞与10 nmol/L的ET-1在37℃下分别培养2、4、6、16和24 h,以不加ET-1培养的细胞为对照,检测各组细胞上清液中PGE2水平的变化。结果：bEnd.3细胞经不同浓度ET-1处理不同时间后,COX-2蛋白表达在2-4h时显著增加,6 h时达到高峰,具有浓度及时间依赖性（F浓度=127.079,F时间=40.158,F交互=5.783,P均〈0.001）。10 nmol/L的ET-1处理不同时间后各组bEnd.3细胞上清液中PGE2释放水平于4 h开始升高,6 h达峰,24 h下降,差异有统计学意义（F=195.630,P〈0.001）。结论：ET-1可上调bEnd.3细胞中炎症因子COX-2的表达及PGE2的释放,最终对bEnd.3细胞株造成损伤。

Aim： To explore the effects of endothelin-1（ ET-1） on the expression of cyclooxygenase-2（ COX-2） and prostaglandin E2（ PGE2） in the mice brain microvascular endothelial cell. Methods： The bEnd. 3 cells,a mice brain microvascular endothelial cell line,were added to 1,10,100 or 1 000 nmol /L ET-1 in treatment for 2,4,6,16 or 24 h. Different concentrations ET-1 treated with different time COX-2 expression was detected by Western blot; another well-grown cells and 10 nmol /L ET-1 were cultured 2,4,6,16 and 24 h at 37 ℃,the cultured cells without ET-1 treatment was used as control. The changes of PGE2 levels in culture supernatants were detected in each group. Results： Compared with control group,the protein expressions of COX-2 were significantly increased at 2 to 4 h and reached the peak at 6 h,with a concentration- and time-dependent manner（ Fconcentration= 127. 079,Ftime= 40. 158,Finteraction= 5. 783,P〈0. 001）; the level of PGE2 were significantly increased at 4 h,reached the peak at 6 h,then were significantly decreased at 24 h after being treated by 10 nmol /L ET-1 in each group bEnd. 3 cell supernatant at different times（ F = 195. 630,P〈0. 001）. Conclusion： ET-1 may up-regulate COX-2 expression and PGE2 release in bEnd. 3 cells,ultimately cause damage to bEnd. 3 cell lines.