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目的观察桂枝汤中分离获得的单体成分桂皮醇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终浓度的桂皮醇(0.093,0.186,0.372,0.744,1.488 mmol.L-1)孵育3 h,之后以30μg.L-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,COX-2的活性。结果Ⅷ因子抗体免疫组化染色可见95%的培养细胞呈阳性,确认为rCMEC。暴露于质量浓度为30μg.L-1IL-1后,rCMEC内COX-2活性及释放的PGE2量显著增加(P<0.01),COX-1活性变化无统计学差异(P>0.05)。加入不同浓度的桂皮醇后,随浓度增加可下调COX-1、COX-2活性及PGE2量,且呈浓度依赖关系;至浓度为0.744mmol.L-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(P<0.05),COX-1活性虽有所降低,但无统计学上的显著差异(P>0.05)。结论桂皮醇能下调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 effects of cinnamyl alcohol isolated from Guizhi Decoction on heat-related cyclooxygenase (COX) and prostaglandin E2 (PGE2). Methods The rat brain microvascular endothelial cells (rCMEC) were cultured and the relevant factor VIII antigens were identified. After the cells were grown to the confluent state, different concentrations of cinnamyl alcohol (0.093, 0.186, 0.372, 0.744, 1.488 mmol.L-1) were added and incubated for 3 h, followed by stimulation with 30 μg.L-1 of IL-1 for 12 h. The ELISA method was used to detect the content of PGE2 in cell culture fluid and the activity of intracellular COX-1 and COX-2. Results VIII factor antibody immunohistochemical staining showed that 95% of cultured cells were positive and confirmed as rCMEC. After exposure to 30μg.L-1IL-1, the activity of COX-2 and the amount of released PGE2 in rCMEC were significantly increased (P<0.01). There was no significant difference in the activity of COX-1 (P>0.05). After adding different concentrations of cinnamic alcohol, COX-1, COX-2 activity, and PGE2 levels were down-regulated with increasing concentrations, and were concentration-dependent; when the concentration was 0.744 mmol.L-1, COX-2 activity and released PGE2 Compared with the IL-1 alone group, there was a significant difference (P <0.05). Although the COX-1 activity was reduced, there was no statistically significant difference (P> 0.05). CONCLUSION: Cinnamyl alcohol can down-regulate IL-6-induced PGE2 release from rCMEC, and the mechanism may be related to the inhibition of COX-2 activity.