分泌型白细胞蛋白酶抑制剂对香烟烟雾提取物诱导人支气管上皮细胞炎症介质表达的影响

目的 检测分泌型白细胞蛋白酶抑制剂(SLPI)对香烟烟雾提取物(CSE)诱导体外培育的正常人支气管上皮细胞(NHBE)炎症介质表达水平的影响,探讨SLPI对慢性气道炎症局部保护作用的机制.方法 实验设对照组、CSE组、SLPI组和SLPI+CSE组,均采用免疫细胞化学方法检测NHBE细胞中基质金属蛋白酶-9(MMP-9)蛋白的表达,采用酶联免疫吸附试验检测各组培养上清液中白细胞介素8(IL-8)的表达.应用SPSS 11.5软件进行统计学处理,多样本均数比较采用单因素方差分析和SNK检验,若方差不齐进行校正,P<0.05为差异有统计学意义.结果 对照组能少量表达MMP-9(积分值为3.1±0.5)和IL-8[(4.9±0.6)ng/L];用CSE干预NHBE细胞能诱导MMP-9和IL-8的表达,且在一定范围内与CSE干预时间呈依赖性,24 h达峰值,MMP-9积分值和IL-8浓度分别为6.6±0.4和(17.7±1.9)ng/L,随后呈明显下降趋势;用SLPI干预NHBE细胞能明显抑制MMP-9(积分值为0.8±0.5)和IL-8[(0.7±0.6)ng/L]的表达.结论 SLPI能抑制CSE诱导的NHBE细胞的MMP-9和IL-8表达.

Effects of secretory leukocyte protease inhibitor on expression of inflammation mediators in normal human bronchial epithelial cells induced by cigarette smoke extract

Objective To study the effect of secretory leukocyte protease inhibitor(SLPT)on the expression of MMP-9 and IL-8 in normal human bronchial epithelial(NHBE)cells induced by cigarette smoke extract(CSE),and therefore to explore the mechanisms of SLPI for protecting the local airways of chronic inflammatory diseases.Methods The experiments of cultured airway epithelia cells in vovo were randomly divided into 4 groups,including a control group,a CSE group,a SLPl group,and a SLPI+CSE group.The expression level of IL-8 in NHBE cell supematant was examined by ELISA.The expression level of MMP-9 protein in NHBE cells was evaluated by using immunocytochemical stain methed.One way analysis of variance was employed in significance test of different groups,followed by SNK test with equal variances and Dunnett3 test with unequal variances.Results A small quantities of MMP-9 and IL-8 expression were observed in the control group NHBE cells.The mean integral expression of MMP-9 protein was(3.1±0.5),and the concentration of IL-8 in NHBE cell supernatant was(4.9±0.6)he/L. After exposure to CSE for different times,the expression of MMP-9 and IL-8 in NHBE cells of the CSE group was higher than those of in control group.The expression levels of MMP-9 and IL-8 were dependent on CSE exposure time Within certain limits.The hishest expression was observed at the time of 24 h exposure to CSE.The mean integral expression of MMP-9 protein wag 6.6±0.4,and the concentration of IL-8 in NHBE cell supernatant wag(17.7±1.9)ng/L. But subsequently the levels decreased significantly in 36 h. When NHBE cells were exposed to 10 μgL SLPI,the expression levels of MMP-9 and IL-8 were inhibited.The integral expression of MMP-9 protein was 0.8±0.5,and the concentration of IL-8 in NHBE cell supernatant was(0.7±0.6)ng/L.Conclusion SLPI inhibited the expression of MMP-9 and IL-8 in NHBE cells induced by cigarette smoking extract.