This study examined the effects of retinoic acid (RA), PD98059, SP600125 and SB203580 on the hyperoxia-induced expression and regulation of matrix metalloproteinase-2 (MMP-2) and metalloproteinase-2 (TIMP-2) in ...This study examined the effects of retinoic acid (RA), PD98059, SP600125 and SB203580 on the hyperoxia-induced expression and regulation of matrix metalloproteinase-2 (MMP-2) and metalloproteinase-2 (TIMP-2) in premature rat lung fibroblasts (LFs). LFs were exposed to hyperoxia or room air for 12 h in the presence of RA and the kinase inhibitors PD98059 (ERK1/2), SP600125 (JNK1/2) and SB203580 (p38) respectively. The expression levels of MMP-2 and TIMP-2 mRNA were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). MMP-2 activity was measured by zymography. The amount of p-ERK1/2, REK1/2, p-JNK1/2, JNK1/2, p-p38 and p38 was determined by Western blotting. The results showed that: (1) PD98059, SP600125 and SB203580 significantly inhibited p-ERK1/2, p-JNK1/2 and p-p38 respectively in LFs; (2) The expression of MMP-2 mRNA in LFs exposed to hyperoxia was decreased after treatment with RA, SP600125 and SB203580 respectively (P0.01 or 0.05), but did not change after treatment with PD98059 (P0.05). Meanwhile, RA, PD98059, SP600125 and SB203580 had no effect on the expression of TIMP-2 mRNA in LFs exposed to room air or hyperoxia (P0.05); (3) The expression of pro- and active MMP-2 experienced no change after treatment with RA or SP600125 in LFs exposed to room air (P0.05), but decreased remarkably after hyperoxia (P0.01 or 0.05). SB203580 inhibited the expression of pro- and active MMP-2 either in room air or under hyperoxia (P0.01). PD98059 exerted no effect on the expression of pro- and active MMP-2 (P0.05). It was suggested that RA had a protective effect on hyperoxia-induced lung injury by down-regulating the expression of MMP-2 through decreasing the JNK and p38 activation in hyperoxia.展开更多
目的探讨脂肪酸结合蛋白5(fatty acid-binding protein 5,FABP5)与细胞维甲酸结合蛋白2(cellular retinoic acid-binding protein 2,CRABP2)在脑胶质瘤中的表达及与脑胶质瘤病理级别、维甲酸治疗抵抗的关系。方法应用免疫组织化学染色...目的探讨脂肪酸结合蛋白5(fatty acid-binding protein 5,FABP5)与细胞维甲酸结合蛋白2(cellular retinoic acid-binding protein 2,CRABP2)在脑胶质瘤中的表达及与脑胶质瘤病理级别、维甲酸治疗抵抗的关系。方法应用免疫组织化学染色法检测125例脑星型细胞瘤组织中FABP5及CRABP2蛋白表达。全反式维甲酸作用前后,以Brdu-ELISA法检测胶质瘤细胞增殖,并以实时荧光定量PCR检测胶质瘤细胞株中FABP5及CRABP2在m RNA水平的表达。结果 FABP5阳性细胞比例在WHOⅡ级星型细胞瘤中占(16±9)%,Ⅲ级中占(33±22)%,Ⅳ级中占(50±29)%,FABP5蛋白表达与胶质瘤病理级别呈显著正相关(P<0.05)。CRABP2阳性细胞比例在Ⅱ级星型细胞瘤中占(46±12)%,WHOⅢ级中占(30±15)%,Ⅳ级中占(10±9)%,CRABP2蛋白表达与胶质瘤病理级别呈显著负相关(P<0.05)。全反式维甲酸促进了胶质瘤细胞株的增殖,全反式维甲酸作用后胶质瘤细胞FABP5 m RNA表达显著上调,而CRABP2显著下调。结论 FABP5及CRABP2的异常表达与胶质瘤恶性程度相关,并可能介导了胶质瘤细胞对维甲酸的分化抵抗效应。展开更多
基金supported by a grant from the Nature Sciences Foundation of China (No. 30872795)
文摘This study examined the effects of retinoic acid (RA), PD98059, SP600125 and SB203580 on the hyperoxia-induced expression and regulation of matrix metalloproteinase-2 (MMP-2) and metalloproteinase-2 (TIMP-2) in premature rat lung fibroblasts (LFs). LFs were exposed to hyperoxia or room air for 12 h in the presence of RA and the kinase inhibitors PD98059 (ERK1/2), SP600125 (JNK1/2) and SB203580 (p38) respectively. The expression levels of MMP-2 and TIMP-2 mRNA were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). MMP-2 activity was measured by zymography. The amount of p-ERK1/2, REK1/2, p-JNK1/2, JNK1/2, p-p38 and p38 was determined by Western blotting. The results showed that: (1) PD98059, SP600125 and SB203580 significantly inhibited p-ERK1/2, p-JNK1/2 and p-p38 respectively in LFs; (2) The expression of MMP-2 mRNA in LFs exposed to hyperoxia was decreased after treatment with RA, SP600125 and SB203580 respectively (P0.01 or 0.05), but did not change after treatment with PD98059 (P0.05). Meanwhile, RA, PD98059, SP600125 and SB203580 had no effect on the expression of TIMP-2 mRNA in LFs exposed to room air or hyperoxia (P0.05); (3) The expression of pro- and active MMP-2 experienced no change after treatment with RA or SP600125 in LFs exposed to room air (P0.05), but decreased remarkably after hyperoxia (P0.01 or 0.05). SB203580 inhibited the expression of pro- and active MMP-2 either in room air or under hyperoxia (P0.01). PD98059 exerted no effect on the expression of pro- and active MMP-2 (P0.05). It was suggested that RA had a protective effect on hyperoxia-induced lung injury by down-regulating the expression of MMP-2 through decreasing the JNK and p38 activation in hyperoxia.
文摘目的探讨脂肪酸结合蛋白5(fatty acid-binding protein 5,FABP5)与细胞维甲酸结合蛋白2(cellular retinoic acid-binding protein 2,CRABP2)在脑胶质瘤中的表达及与脑胶质瘤病理级别、维甲酸治疗抵抗的关系。方法应用免疫组织化学染色法检测125例脑星型细胞瘤组织中FABP5及CRABP2蛋白表达。全反式维甲酸作用前后,以Brdu-ELISA法检测胶质瘤细胞增殖,并以实时荧光定量PCR检测胶质瘤细胞株中FABP5及CRABP2在m RNA水平的表达。结果 FABP5阳性细胞比例在WHOⅡ级星型细胞瘤中占(16±9)%,Ⅲ级中占(33±22)%,Ⅳ级中占(50±29)%,FABP5蛋白表达与胶质瘤病理级别呈显著正相关(P<0.05)。CRABP2阳性细胞比例在Ⅱ级星型细胞瘤中占(46±12)%,WHOⅢ级中占(30±15)%,Ⅳ级中占(10±9)%,CRABP2蛋白表达与胶质瘤病理级别呈显著负相关(P<0.05)。全反式维甲酸促进了胶质瘤细胞株的增殖,全反式维甲酸作用后胶质瘤细胞FABP5 m RNA表达显著上调,而CRABP2显著下调。结论 FABP5及CRABP2的异常表达与胶质瘤恶性程度相关,并可能介导了胶质瘤细胞对维甲酸的分化抵抗效应。