Summary:To examine the mRNA expression of matrix metalloproteinase 9 (MMP-9) and the gelatinase activity of its inhibitor, tissue inhibitor of matrix metalloproteinase 1 (TIMP-1) in the primary epithelial cells of pat...Summary:To examine the mRNA expression of matrix metalloproteinase 9 (MMP-9) and the gelatinase activity of its inhibitor, tissue inhibitor of matrix metalloproteinase 1 (TIMP-1) in the primary epithelial cells of patients with COPD, airway epithelial cells were taken from 15 COPD patients and cultured in vitro. The patients were divided into three groups, COPD group, normal smoking control group and non-smoking control group, with 5 subjects in each group, on basis of the smoking history and lung function. The semi-qualitative RT-PCR was employed to determine the mRNA levels of MMP-9 and TIMP-1 and SDS PAGE was used for the determination of the gelatinase activity of MMP-9 and TIMP-1. Our result showed that the mRNA of MMP-9 and TIMP-1 in epithelial cells of the non-smoking subjects was at a low level. The mRNA of MMP-9 and TIMP-1 in COPD patients and smokers was significantly higher than that in non-smokers (P<0.05). No significant difference was found in the levels of MMP-9 and TIMP-1 in epithelial cells between the COPD patients and smokers. The MMP-9/TIMP-1 ratios in COPD patients and smokers were significantly lower than that of non-smokers (P<0.05). The gelatinase activity in the epithelial cells of both COPD patients and normal smokers was increased (P<0.05), but no difference existed in the gelatinase activity in the epithelial cells between COPD patients and normal smokers. It is concluded that the transcription of MMP-9 and TIMP-1 and the gelatinase activity of MMP-9 and MMP-2 in the epithelial cells in COPD patients were increased, which resulted in an imbalance of MMP-9/TIMP-1, thereby causing pulmonary fibrosis. These factors play important roles in the pathogenesis of COPD.展开更多
INTRODUCTIONOur lungs have a vital role in mediating the exchange of oxygen and carbon dioxide between the air we breathe and the body.This function is under constant pressure as inhaled air contains numerous particle...INTRODUCTIONOur lungs have a vital role in mediating the exchange of oxygen and carbon dioxide between the air we breathe and the body.This function is under constant pressure as inhaled air contains numerous particles,gasses,and microorganisms that may cause injury and infection to the lungs.Removal and neutralization of potential harmful substances from inhaled air is a main function of the airway epithelium.展开更多
目的:揭示miR-455-5p对呼吸道合胞病毒(RSV)感染致气道上皮细胞炎症反应的作用机制。方法:qRT-PCR检测30例健康体检儿童(健康组)、RSV感染轻症组(n=41)和重症组(n=31)患儿血清miR-455-5p水平。将16HBE细胞分为Control组、NC-agomir组、m...目的:揭示miR-455-5p对呼吸道合胞病毒(RSV)感染致气道上皮细胞炎症反应的作用机制。方法:qRT-PCR检测30例健康体检儿童(健康组)、RSV感染轻症组(n=41)和重症组(n=31)患儿血清miR-455-5p水平。将16HBE细胞分为Control组、NC-agomir组、miR-455-5p-agomir组、NC-antagomir组、miR-455-5p-antagomir组。使用Lipofectamine 3000转染16HBE细胞后培养48 h后,分为Blank组、NC组(转染了NC-agomir的细胞)、RSV+NC-agomir组、RSV+miR-455-5p-agomir组,用RSV病毒液感染RSV+NC-agomir组和RSV+miR-455-5p-agomir组16HBE细胞,Blank组和NC组16HBE细胞正常培养。用CCK-8法和EdU法检测细胞增殖、TUNEL法检测细胞凋亡、ELISA法检测上清液中TNF-α、IL-6、IL-8水平,qRT-PCR检测miR-455-5p和SOCS3的m RNA水平,Western blot检测SOCS3、IFN-α、STAT1、STAT2、p-STAT1和p-STAT2的蛋白水平。结果:与健康组相比,轻症组和重症组患儿的血清miR-455-5p水平降低(P<0.05)。与轻症组相比,重症组的血清miR-455-5p水平降低(P<0.05)。与Control组和NC-agomir组相比,miR-455-5p-agomir组16HBE细胞的miR-455-5p水平、相对细胞活力和EdU阳性率升高(P<0.05),TUNEL阳性率降低(P<0.05),上清液中的TNF-α、IL-6和IL-8水平降低(P<0.05),SOCS3 m RNA和蛋白水平降低(P<0.05),IFN-α蛋白、STAT1和STAT2磷酸化水平升高(P<0.05)。与Control组和NC-antagomir组相比,miR-455-5p-antagomir组16HBE细胞的miR-455-5p水平、相对细胞活力和EdU阳性率降低(P<0.05),TUNEL阳性率升高(P<0.05),上清液中的TNF-α、IL-6和IL-8水平升高(P<0.05),SOCS3 m RNA和蛋白水平升高(P<0.05),IFN-α蛋白、STAT1和STAT2磷酸化水平降低(P<0.05)。与Blank组和NC组相比,RSV+NC-agomir组16HBE细胞的miR-455-5p水平、相对细胞活力和EdU阳性率降低(P<0.05),TUNEL阳性率升高(P<0.05),上清液中的TNF-α、IL-6和IL-8水平升高(P<0.05),SOCS3 m RNA和蛋白水平升高(P<0.05),IFN-α蛋白、STAT1和STAT2磷酸化水平降低(P<0.05)。与RSV+NC-agomir组相比,RSV+miR-455-5p-展开更多
文摘Summary:To examine the mRNA expression of matrix metalloproteinase 9 (MMP-9) and the gelatinase activity of its inhibitor, tissue inhibitor of matrix metalloproteinase 1 (TIMP-1) in the primary epithelial cells of patients with COPD, airway epithelial cells were taken from 15 COPD patients and cultured in vitro. The patients were divided into three groups, COPD group, normal smoking control group and non-smoking control group, with 5 subjects in each group, on basis of the smoking history and lung function. The semi-qualitative RT-PCR was employed to determine the mRNA levels of MMP-9 and TIMP-1 and SDS PAGE was used for the determination of the gelatinase activity of MMP-9 and TIMP-1. Our result showed that the mRNA of MMP-9 and TIMP-1 in epithelial cells of the non-smoking subjects was at a low level. The mRNA of MMP-9 and TIMP-1 in COPD patients and smokers was significantly higher than that in non-smokers (P<0.05). No significant difference was found in the levels of MMP-9 and TIMP-1 in epithelial cells between the COPD patients and smokers. The MMP-9/TIMP-1 ratios in COPD patients and smokers were significantly lower than that of non-smokers (P<0.05). The gelatinase activity in the epithelial cells of both COPD patients and normal smokers was increased (P<0.05), but no difference existed in the gelatinase activity in the epithelial cells between COPD patients and normal smokers. It is concluded that the transcription of MMP-9 and TIMP-1 and the gelatinase activity of MMP-9 and MMP-2 in the epithelial cells in COPD patients were increased, which resulted in an imbalance of MMP-9/TIMP-1, thereby causing pulmonary fibrosis. These factors play important roles in the pathogenesis of COPD.
文摘INTRODUCTIONOur lungs have a vital role in mediating the exchange of oxygen and carbon dioxide between the air we breathe and the body.This function is under constant pressure as inhaled air contains numerous particles,gasses,and microorganisms that may cause injury and infection to the lungs.Removal and neutralization of potential harmful substances from inhaled air is a main function of the airway epithelium.
文摘目的:揭示miR-455-5p对呼吸道合胞病毒(RSV)感染致气道上皮细胞炎症反应的作用机制。方法:qRT-PCR检测30例健康体检儿童(健康组)、RSV感染轻症组(n=41)和重症组(n=31)患儿血清miR-455-5p水平。将16HBE细胞分为Control组、NC-agomir组、miR-455-5p-agomir组、NC-antagomir组、miR-455-5p-antagomir组。使用Lipofectamine 3000转染16HBE细胞后培养48 h后,分为Blank组、NC组(转染了NC-agomir的细胞)、RSV+NC-agomir组、RSV+miR-455-5p-agomir组,用RSV病毒液感染RSV+NC-agomir组和RSV+miR-455-5p-agomir组16HBE细胞,Blank组和NC组16HBE细胞正常培养。用CCK-8法和EdU法检测细胞增殖、TUNEL法检测细胞凋亡、ELISA法检测上清液中TNF-α、IL-6、IL-8水平,qRT-PCR检测miR-455-5p和SOCS3的m RNA水平,Western blot检测SOCS3、IFN-α、STAT1、STAT2、p-STAT1和p-STAT2的蛋白水平。结果:与健康组相比,轻症组和重症组患儿的血清miR-455-5p水平降低(P<0.05)。与轻症组相比,重症组的血清miR-455-5p水平降低(P<0.05)。与Control组和NC-agomir组相比,miR-455-5p-agomir组16HBE细胞的miR-455-5p水平、相对细胞活力和EdU阳性率升高(P<0.05),TUNEL阳性率降低(P<0.05),上清液中的TNF-α、IL-6和IL-8水平降低(P<0.05),SOCS3 m RNA和蛋白水平降低(P<0.05),IFN-α蛋白、STAT1和STAT2磷酸化水平升高(P<0.05)。与Control组和NC-antagomir组相比,miR-455-5p-antagomir组16HBE细胞的miR-455-5p水平、相对细胞活力和EdU阳性率降低(P<0.05),TUNEL阳性率升高(P<0.05),上清液中的TNF-α、IL-6和IL-8水平升高(P<0.05),SOCS3 m RNA和蛋白水平升高(P<0.05),IFN-α蛋白、STAT1和STAT2磷酸化水平降低(P<0.05)。与Blank组和NC组相比,RSV+NC-agomir组16HBE细胞的miR-455-5p水平、相对细胞活力和EdU阳性率降低(P<0.05),TUNEL阳性率升高(P<0.05),上清液中的TNF-α、IL-6和IL-8水平升高(P<0.05),SOCS3 m RNA和蛋白水平升高(P<0.05),IFN-α蛋白、STAT1和STAT2磷酸化水平降低(P<0.05)。与RSV+NC-agomir组相比,RSV+miR-455-5p-
