摘要
目的探讨TNFΑ对慢性阻塞性肺疾病(COPD)患者肺泡巨噬细胞(AM)源性基质金属蛋白酶9(MMP-9)表达与酶活性的影响及其可能调节机制。方法从COPD患者支气管肺泡灌洗液中分离与培养AM,以0.1、1、5、10、20 NG/M L TNFΑ刺激24 H后,用半定量RT-PCR法检测MMP-9MRNA表达,明胶酶谱法检测MMP-9酶活性,凝胶阻滞分析实验检测NF-ΚB活性。结果当AM未被TNFΑ刺激时,MMP-9 MRNA表达吸光度比值为0.1826±0.0084,MMP-9酶活性的相对吸光度值为10 227±738。随着TNFΑ刺激剂量增加,MMP-9 MRNA表达相应增加,分别为0.4279±0.0204、0.5431±0.0058、0.6185±0.0208、0.6859±0.0080、0.7784±0.0208(P<0.05);MMP-9酶活性相应增强,分别为40 528±974、113 741±4547、190 696±4032、309 505±7639、427 305±9145(P<0.05)。以10 NG/M L TNFΑ刺激AM后,NF-ΚB的活性较刺激前显著增强(P<0.05)。结论TNFΑ能够诱导COPD患者AM表达MMP-9,TNFΑ诱导的AM源性MMP-9的表达可能与NF-ΚB介导的信号转导途径有关。
Objective To explore the expression and activity of matrix metalloproteinase-9 （MMP-9） from alveolar macrophages （AMs） induced by tumor necrosis factor-or （TNFα） in patients with chronic obstructive pulmonary disease （COPD）, and therefore to study the possible mechanisms. Methods AMs were collected from bronchoalveolar lavage fluid from patients with COPD, and then were stimulated for 24 h with TNFα at concentrations of 0. 1 ng/ml, 1 ng/ml, 5 ng/ml, 10 ng/ml,and 20 ng/ml, respectively. The gene expression level of MMP-9 induced by TNFα was detected by semi-quantitative RT-PCR, and MMP-9 activity was measured by zymography. NF-KB activity was detected by electrophoretic mobility shift assay. Results In unstimulated AMs, the densitometry unit rate of MMP-9 mRNA to 13-actin mRNA was 0. 1826 ±0. 0084. When AMs were stimulated by TNFα at concentrations of 0. 1 ng/ml, 1 ng/ml, 5 ng/ml, 10 ng/ml, and 20 ng/ml, the densitometry unit rates of MMP-9 mRNA to β-actin mRNA were 0. 4279 ± 0. 0204, 0. 5431 ±0. 0058, 0. 6185 ±0. 0208, 0. 6859 ±0. 0080, and 0. 7784 ±0. 0208, respectively. In unstimulated AMs, the densitometry unit of active MMP-9 was 10 227 ± 738,but when AMs were stimulated by TNFα at concentrations of 0. 1 ng/ml, 1 ng/ml, 5 ng/ml, 10 ng/ml, and 20 ng/ml, the densitometry units of active MMP-9 were 40 528 ± 974, 113 741 ± 4547, 190 696 ± 4032, 309 505 ± 7639, and 427 305 ±9145, respectively. NF-KB activity was significantly elevated after AMs were stimulated by TNFα at the concentration of 10 ng/ml （P 〈 0. 05）. Conclusions The expression and activity of MMP-9 can be induced by TNFα, and the induction was possibly related with the NF-KB-mediated signal transduction pathway.
出处
《中华内科杂志》
CAS
CSCD
北大核心
2006年第1期38-41,共4页
Chinese Journal of Internal Medicine
