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目的:观察桂枝汤有效部位A分离的邻甲氧基桂皮醛对发热相关的环氧酶(COX)和前列腺素E2(PGE2)的影响。方法:建立大鼠脑微血管内皮细胞(rCMEC)培养,进行Ⅷ因子相关抗原鉴定。待细胞生长至融合状态后加入不同含量的邻甲氧基桂皮醛(12.5,25,50,100,200μg.mL-1)孵育3 h,之后以30 ng.mL-1的IL-1刺激12 h。ELISA方法检测细胞培养液中PGE2的含量及细胞内COX-1和COX-2的活性。结果:Ⅷ因子抗体免疫组化染色可见90%以上的培养细胞呈阳性,确认为rCMEC。暴露于30 ng.mL-1IL-1后,rCMEC内COX-2活性及释放的PGE2量显著增加,COX-1活性变化无统计学差异。加入不同含量的邻甲氧基桂皮醛后,随含量增加可下调COX-1,COX-2活性及PGE2量,且呈剂量依赖关系;至含量为200μg.mL-1时,COX-2活性及释放的PGE2与IL-1单独作用组相比均有显著性差异,COX-1活性虽有所降低,但无统计学上的显著差异。结论:邻甲氧基桂皮醛能下调IL-1刺激rCMEC释放升高的PGE2,作用机制可能与抑制COX-2活性有关。
OBJECTIVE: To observe the effect of o-methoxycinnamic aldehyde on the heat-related cyclooxygenase (COX) and prostaglandin E2 (PGE2) in the effective part A of Guizhi decoction. METHODS: Rat brain microvascular endothelial cells (rCMEC) were cultured to identify factor VIII related antigens. After the cells were grown to fused state, different concentrations of o-methoxycinnamic aldehyde (12.5, 25, 50, 100, 200 μg.mL-1) were added for 3 hours, followed by 30 ng.mL-1 of IL-1 for 12 hours. The content of PGE2 in cell culture fluid and the activity of COX-1 and COX-2 in cells were detected by ELISA. RESULTS: Immunohistochemical staining for factor VIII showed that more than 90% of cultured cells were positive and confirmed as rCMEC. After exposure to 30 ng.mL-1IL-1, the activity of COX-2 and the release of PGE2 were significantly increased in rCMEC, and there was no significant difference in the activity of COX-1. After adding different contents of o-methoxycinnamic aldehyde, the COX-1, COX-2 activity and PGE2 content were decreased with the increase of the content in a dose-dependent manner; when the content was 200 μg.mL-1, COX-2 activity and There was a significant difference in the released PGE2 compared with the IL-1 alone group. Although the COX-1 activity was reduced, there was no statistically significant difference. CONCLUSION: O-methoxycinnamic aldehyde can down-regulate IL-6-induced PGE2 release from rCMEC, and the mechanism may be related to the inhibition of COX-2 activity.