薯蓣皂苷元对白细胞的粘附与迁移及内皮细胞ICAM-1的表达和NF-κB活化的影响(英文)  

Diosgenin reduces leukocytes adhesion and migration linked with inhibition of intercellular adhesion molecule-1 expression and NF-κB p65 activation in endothelial cells

作　　者：宋佳希[1,2,3] 马丽 寇俊萍[1,2] 余伯阳[1,2] 

机构地区：[1]中国药科大学中药复方研究室,南京211198 [2]天然药物活性组分与药效国家重点实验室中国药科大学,南京211198 [3]南京军区南京总医院解放军临床检验医学研究所

出　　处：《中国天然药物》2012年第2期142-149,共8页

基　　金：supported by the Major Research Plan of the National Natural Science Foundation of China (No.90713042);Program for New-Century Excellent Talents in University (NCET-07-849);the Priority Academic Program Development of Jiangsu Higher Education Institutions~~

摘　　要：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。AIM： To investigate the anti-inflammatory activity and possible mechanism of diosgenin, and to provide some further pharmacological evidence for its clinical applications. METHODS： Anti-inflammatory activities of diosgenin were evaluated by zymosan A-evoked peritoneal leukocytes migration in mice and the adhesion of human pro-myelocytic leukemia cell strain （HL-60） to human umbilical vein endothelial cell line （ECV304） cells induced by tumor necrosis factor （TNF-α）. Furthermore, the effects of diosgenin on TNF-α-induced intercellular adhesion molecule-1 （ICAM-1） expression were also investigated by reverse transcrip-tion-PCR （RT-PCR） and flow cytometric analysis, and nuclear factor-kappaB （NF-κB）/p65 translocation and phosphorylation were determined by Western blot. RESULTS： Diosgenin significantly inhibited peritoneal leukocytes migration induced by zymosan A in mice when given orally once at doses of 1 and 3 mg·kg-1. Pretreatment ECV304 cells with diosgenin at concentrations of 0.1 and 1 mol·L？1 for 5 h remarkably decreased TNF-α-stimulated adhesion of HL-60 to ECV304 cells in vitro, with no effect on viability of ECV304 cells. Furthermore, diosgenin inhibited TNF-α-induced overexpression of ICAM-1 both at the mRNA and protein levels, and suppressed nuclear p65 accumulation and p65 activation in ECV304 cells. CONCLUSION： These results suggested that diosgenin significantly inhibited leukocytes migration and adhesion, partly due to the down-regulation of ICAM-1 expression through NF-κB path-way.

关 键 词：薯蓣皂苷元 白细胞粘附和迁移 细胞间粘附因子-1 核因子--κB 

分 类 号：R965[医药卫生—药理学]