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目的:研究桂枝汤及其生物转化模拟产物对白细胞介素-1β(IL-1β)介导的小鼠脑微血管内皮细胞株(bEnd.3)细胞前列腺素E2(PGE2)代谢通路的影响。方法:制备了桂枝汤及桂枝汤生物转化模拟产物的含药血清(分别设五个浓度组),采用酶联免疫吸附法(ELISA)和免疫印记法(Western blot),观察桂枝汤及其生物转化模拟产物对IL-1β刺激的bEnd.3细胞磷酸化丝裂原活化的蛋白激酶p38(p-p38MAPK)、胞浆型磷脂酶A2(cPLA2)、分泌型磷脂酶A2(sPLA2)、环氧合酶2(COX-2)、前列腺素E合成酶(PGES)、15-羟基前列腺素脱氢酶(15-PGDH)、磷酸化核因子κB(p-NF-κB)以及前列腺素E2(PGE2)分泌及表达的影响。结果:IL-1β作用于细胞10 h后,各项指标均明显上调,与对照组比较,P<0.01。除质量终浓度为2.5%的桂枝汤含药血清对PGES,sPLA2;终浓度为1%的模拟药对PGES;终浓度为2.5%的含药血清对PGES,sPLA2,cPLA2作用不显著外,其他各浓度组与模型组比较均出现显著性差异。结论:桂枝汤及其生物转化模拟产物对IL-1β刺激bEnd.3细胞合成PGE2的代谢酶系统有不同程度的抑制作用,从而降低了前炎症介质和中枢发热介质PGE2的释放和分泌。结果提示,桂枝汤及其生物转化模拟产物对脑微血管内皮细胞分泌PGE2具有相似的药效,为模拟产物的应用和开发提供了实验支持。
Objective: To study the effect of Guizhi decoction and its biotransformation analogue on prostaglandin E2 (PGE2) pathway in mouse brain microvascular endothelial cell line (bEnd.3) mediated by interleukin-1β (IL-1β). Methods: The medicated serum of Guizhi decoction and Guizhi decoction (five concentrations group) were prepared. Enzyme linked immunosorbent assay (ELISA) and Western blot were used to observe the effects of Gui Zhi Tang (P-p38MAPK), cytoplasmic phospholipase A2 (cPLA2), secreted phospholipase A2 (sPLA2) and phosphorylated mitogen-activated protein kinase , Cyclooxygenase 2 (COX-2), prostaglandin E synthase (PGES), 15-hydroxyprostaglandin dehydrogenase (15-PGDH), phosphorylated nuclear factor κB (p-NF-κB) and prostaglandin E2 (PGE2) secretion and expression. Results: After treated with IL-1β for 10 h, all the indexes were significantly increased, P <0.01 compared with the control group. In addition to PGES and sPLA2, the drug-containing serum of Guizhi decoction with the final concentration of 2.5% had no significant effect on PGES, sPLA2 and cPLA2 at the final concentration of 1% Other concentrations of each group compared with the model group showed significant differences. CONCLUSION: Guizhi Decoction and its biotransformation analogue products can inhibit the metabolism of enzyme system of PGE2 stimulated by IL-1β to a certain extent, thereby reducing the release and secretion of PGE2 in pro-inflammatory and central febrile media. The results suggest that Guizhi Decoction and its biotransformation analogue have similar efficacy on the secretion of PGE2 by brain microvascular endothelial cells and provide experimental support for the application and development of simulated products.