5-氨基水杨酸诱导体外人中性粒细胞凋亡

Induction of apoptosis of human neutrophil by 5 aminosalicylic acid in vitro

目的观察５氨基水杨酸（５ＡＳＡ）诱导体外中性粒细胞（ｐｏｌｙｍｏｒｐｈｏｎｕｃｌｅａｒｎｅｕｔｒｏｐｈｉｌ，ＰＭＮ）凋亡的作用。方法Ｆｉｃｏｌ离心法分离健康人ＰＭＮ和单个核细胞。分别用５ＡＳＡ和（或）脂多糖（ＬＰＳ）、放线菌酮（ＣＨＸ）处理ＰＭＮ及单个核细胞。细胞凋亡判断用形态学方法（Ｇｉｅｍｓａ和吖啶橙染色）和ＤＮＡ电泳法。用吖啶橙染色法在荧光显微镜下行凋亡细胞计数。结果５ＡＳＡ可特异地诱导ＰＭＮ的凋亡，而对周边血单个核细胞没有这种作用。５ＡＳＡ和ＬＰＳ共处理后，ＰＭＮ的凋亡率与对照组和单纯５ＡＳＡ组相比显著增高（Ｐ＜００１）。５ＡＳＡ对ＰＭＮ的凋亡的诱导作用与其作用时间和浓度成平行关系。ＣＨＸ对５ＡＳＡ诱导ＰＭＮ的凋亡作用没有明显影响。结论５ＡＳＡ可以特异地诱导体外ＰＭＮ的凋亡，这至少在部分上可以解释５ＡＳＡ在ＩＢＤ治疗中的作用机制。

Abstract AIM To confirm the hypothesis that therapeutic effects of 5 aminosalicylic acid (5 ASA) on inflammatory bowel diseases were mediated by induction of neutrophils. METHODS Neutrophils and monocytes isolated from healthy human donors were incubated for 12 h in 10% RPMI 1640 medium at 37℃ with 0 05 CO 2. The cells were divided into different subgroups and were incubated with 5 ASA and(or) lipopolysaccharide (LPS, 1 mg·L -1 ), of cycloheximide (5 mg·L -1 ,CHX). Apoptosis was assessed morphologically using Giemsa staining, acridinge orange staining and biochemically by DNA gel electrophoresis. PMN stained with acridine orange and