CAJ | 학술논문

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.
목적 검측분비형백세포단백매억제제(SLPI)대향연연무제취물(CSE)유도체외배육적정상인지기관상피세포(NHBE)염증개질표체수평적영향,탐토SLPI대만성기도염증국부보호작용적궤제.방법 실험설대조조、CSE조、SLPI조화SLPI+CSE조,균채용면역세포화학방법검측NHBE세포중기질금속단백매-9(MMP-9)단백적표체,채용매련면역흡부시험검측각조배양상청액중백세포개소8(IL-8)적표체.응용SPSS 11.5연건진행통계학처리,다양본균수비교채용단인소방차분석화SNK검험,약방차불제진행교정,P<0.05위차이유통계학의의.결과 대조조능소량표체MMP-9(적분치위3.1±0.5)화IL-8[(4.9±0.6)ng/L];용CSE간예NHBE세포능유도MMP-9화IL-8적표체,차재일정범위내여CSE간예시간정의뢰성,24 h체봉치,MMP-9적분치화IL-8농도분별위6.6±0.4화(17.7±1.9)ng/L,수후정명현하강추세;용SLPI간예NHBE세포능명현억제MMP-9(적분치위0.8±0.5)화IL-8[(0.7±0.6)ng/L]적표체.결론 SLPI능억제CSE유도적NHBE세포적MMP-9화IL-8표체.
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.