摘要
目的探索丝氨酸蛋白酶3（PR3）对人外周血树突状细胞（DC）样单核细胞-CD14＋CD16^high单核细胞的蛋白酶活化受体（PAR）-2表达、成熟及细胞因子产生的作用。方法密度梯度离心法分离健康人和韦格纳肉芽肿病（WG）患者外周血单个核细胞（PBMC）；分别以脂多糖、PR3、胰蛋白酶、PAR-2激动肽（PAR-2-AP）、脂多糖＋PR3、脂多糖＋胰蛋白酶在体外刺激PBMC24h；流式细胞仪检测刺激后的CD14＋CD16恸单核细胞表面PAR-2及CD80、CD83、人类白细胞抗原（HLA）-DR的表达；酶联免疫吸附试验（ELISA）检测培养上清中细胞因子白细胞介素（IL）-6的分泌。统计学分析采用MannWhitneY非参数检测方法。结果PR3、胰蛋白酶和PAR-2-AP对外周血DC样单核细胞表面PAR-2和CD80、CD83、HLA—DR表达均无影响；脂多糖能够显著升高健康人细胞表面PAR-2[（5．8±1．5）％升至（24．5±4．5）％，P=0．002]及WG患者和健康人CD80、CD83、HLA—DR的表达；PR3、胰蛋白酶、PAR-2-AP及脂多糖均能诱导IL-6的分泌。结论PR3和PAR-2通路激动剂不能诱导DC样单核细胞的PAR-2的表达及成熟，但能诱导IL-6分泌。
Objective To assess the effects of protease 3 （PR3） and protease-activated receptor （PAR）-2-activator on the maturation and functions of peripheral blood dendritic cell （DC）-like monocytes. Methods Density gradient eentrifugation was used to isolate peripheral blood mononuclear cells （PBMC） from Wegener＇s granulomatosis （WG） patients and healthy controls （HC）. PBMC were stimulated by LPS, human PR3, trypsin, PAR-2-agonist peptide （PAR-2-AP）, LPS＋PR3 or LPS＋trypsin for 24 h. Flow cytometry was used to analyze the expression of PAR-2, CD80, CD83, HLA-DR on stimulated DC-like monocytes-CD14＋CD16＋1 monoeytes. ELISA kit was used to test the concentration of IL-6 in the culture supernants. Mann-Whitney non-parameterie test was used for statistical analysis. Results No effect of PR3, trypsin and PAR-2-AP on the expression of PAR-2, CD80, CD83, HLA-DR of PC-like monocytes was found. LPS could significantly induce PAR-2 expression in HC Ifrom （5.8±1.5）% to （24.5±4.5）%, P=0.002] and the expression of CD80, CD83, HLA-DR in HC and WG;PR3, trypsin, PAR-2-AP and LPS could all stimulate the secretion of IL-6. Conclusion PR3 and PAR-2 pathway-activators can not promote PAR-2 expression and maturation of DC-like monoeytes, but they can induce the secretion of IL-6.
出处
《中华风湿病学杂志》
CAS
CSCD
北大核心
2011年第5期341-344,共4页
Chinese Journal of Rheumatology
