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目的:探讨薯蓣皂苷元的抗炎活性及其可能机制,为其临床应用提供药理学依据。方法:采用酵母多糖A诱导的小鼠腹膜腔白细胞游走模型,评价薯蓣皂苷元的体内抗炎活性;利用白细胞与肿瘤坏死因子(TNF-α)活化的内皮细胞粘附的实验,验证薯蓣皂苷元的抗炎活性;采用聚合酶链式反应和流式细胞术,考察内皮细胞细胞间粘附分子-1(ICAM-1)的表达;蛋白印迹法测定细胞核中转录因子NF-κB/p65的表达及细胞裂解液中NF-κB/p65的磷酸化情况,以探讨薯蓣皂苷元抗炎的可能机制。结果:薯蓣皂苷元1,3mg·kg1灌胃给药1次,可显著减少酵母多糖A诱导的小鼠腹腔内白细胞数量;薯蓣皂苷元0.1,1μmol·L1体外加药5h,显著抑制人HL-60细胞与TNF-α活化的ECV304细胞的粘附,但对ECV304细胞活力无明显影响,也不影响HL-60细胞与静息状态ECV304细胞的粘附。同时,薯蓣皂苷元0.1,1μmol·L1明显抑制TNF-α诱导的ECV304细胞ICAM-1mRNA和蛋白的过量表达,1μmol·L1可抑制NF-κB/p65的核转移;薯蓣皂苷元明显减少TNF-α刺激10～30min诱导的NF-κB/p65的磷酸化,并呈一定浓度依赖性。结论:薯蓣皂苷元具有显著抑制白细胞的黏附迁移活性,其部分机制与抑制内皮细胞NF-κB活化,下调ICAM-1的表达有关。
Objective: To investigate the anti-inflammatory activity of diosgenin and its possible mechanism, to provide pharmacological evidence for its clinical application. Methods: The peritoneal leukocyte migration model induced by zymosan A in mice was used to evaluate the anti-inflammatory activity of diosgenin in vivo. The adhesion of leukocytes to tumor necrosis factor (TNF) -activated endothelial cells was tested to verify the effect of diosgenin The expression of ICAM-1 in endothelial cells was detected by polymerase chain reaction and flow cytometry. The expression of NF-κB / p65 Expression and phosphorylation of NF-κB / p65 in cell lysis solution to explore the possible mechanism of diosgenin anti-inflammatory. Results: Diosgenin 1, 3 mg · kg -1 administered orally once could significantly reduce the number of peritoneal white blood cells induced by zymosan A. Diosgenin 0.1 and 1 μmol·L -1 were administered in vitro for 5 h, 60 cells adhered to TNF-α-activated ECV304 cells, but had no significant effect on the viability of ECV304 cells and did not affect the adhesion of HL-60 cells to resting ECV304 cells. At the same time, diosgenin 0.1,1μmol·L1 significantly inhibited the over-expression of ICAM-1mRNA and protein in ECV304 cells induced by TNF-α, 1μmol·L1 inhibited the nuclear translocation of NF-κB / p65; diosgenin significantly reduced TNF-α The phosphorylation of NF-κB / p65 induced by 10 ~ 30min was stimulated in a concentration-dependent manner. CONCLUSION: Diosgenin can significantly inhibit leukocyte adhesion and migration. Its mechanism may be related to the inhibition of NF-κB activation and the down-regulation of ICAM-1 expression.