原发性免疫性血小板减少症患者颗粒溶素、颗粒酶B、穿孔素的表达及临床意义

Expression of Granzyme B,Granulysin and Perforin from Patients with Primary Immune Thrombocytopenia

目的研究颗粒溶素（granulysin，GNLY）、颗粒酶B（granzymeB，GrB）及穿孔素（perforin，PFP）三种免疫分子在原发性免疫性血小板减少症（primaryimmunethromboeytopenia，ITP）患者外周血单个核细胞（peripheralbloodmononucle—arcells，PBMC）中的表达及其临床意义。方法使用实时荧光定量PCR（RFQ_PCR）和免疫印迹法（westernblot，WB）检测30例ITP患者和40例健康人PBMC中GNLY，GrB及PFP的mRNA和蛋白含量。分析ITP患者三种蛋白含量与IL-2，IL-4，IL-10和IFN-γ的相关性。结果与健康对照组比较，ITP患者PBMC中GrB，PFPmRNA（1．78±0．13VS1.0O±0．03；1．96±0．11VS1．O0±0．04）及蛋白含量（2．05±0．19VS1．02±0．10；2．03±0．23VS0．92±0．15）均升高（t=23．016～45．594，P值均〈0．01），而GNLY变化差异无统计学意义（P〉0．05）。ITP患者中GrB，PFP蛋白表达水平与血小板数量呈负相关（r=-0．401，-0．486，P〈0．05），与IL-2（r=0．515，0．565），IFN-γ（r=0．478，0．544）水平呈正相关（P〈0．05），与IL-4，IL-10无相关性（P〉0．05）。结论GrB及PFP可能参与ITP的发病机制，为探讨ITP的病因和治疗提供了新线索。

Objective To investigate the expression of granulysin （GNLY） ,granzyme B （GrB） and perforin （PFP） in periph eral blood mononuclear cells （PBMC） from patients with primary immune thrombocytopenia （ITP） and its clinical signifi eance. Methods The GNLY,GrB and the PFP mRNA and protein exprssion were determined by real-time fluorescent quan- titative PCR （RFQ-PCR） and western blot （western blot） in 30 ITP patients and 40 healthy controls. Analysed the correla tion between the level of the three immune moleculars and IL- 2,IL-4,IL-10 and IFN-y. Results Compared to healthy con- trols,GrB,PFP mRNA （1.78±0. 13 vs 1.00±0.03;1.96±0.11 vs 1.00±0.04） and protein （2.05±0.19 vs 1.02±0. 10; 2.03±0.23 vs 0. 92±0.15） expression from ITP patients were significantly increased （t=23. 016~45. 594,P〈0.01）,and there was no significant difference in GNLY expression （P〉0.05）. GrB,PFP protein expression in ITP patients were nega tively correlated with platelet count （r=-0. 401,-0. 486,P〈0.05） ,positively correlated with IL 2 （r=0. 515,0. 565）, IFN -γ （r=0. 478,0. 544 ,P〈0.05） ,and there was no association wkh IL-4 and IL-10 （P〉0.05）. Conclusion GrB and PFP would be involved in the pathogenesis of ITP. Suggests a new clue for pathogenesis and effective treatment for ITP.