论文部分内容阅读
目的观察不同缺氧时间对肺成纤维细胞结缔组织生长因子(CTGF)和基质金属蛋白酶9(MMP-9)表达的影响。方法原代培养的人肺成纤维细胞在缺氧(37℃,5%CO_2,5%O_2,90%N_2)条件下分别孵育0、1.5、3、6、12h,反转录聚合酶链反应(RT-PCR)法测定细胞CTGF mRNA、MMP-9 mRNA表达情况,并以酶联免疫吸附试验(ELISA)法测定培养细胞上清液CTGF、MMP-9浓度。结果缺氧刺激成纤维细胞CTGF mRNA高表达出现在1.5h,并保持一相对平台至6h,12h开始下降。MMP-9 mRNA在缺氧1.5h出现高表达,并保持一持续增高的趋势。ELISA结果证实CTGF与MMP-9一样其峰浓度在缺氧12 h。用抗CTGF抗体干预的成纤维细胞组较未干预组表达MMP-9量明显减少。结论缺氧可促使人肺成纤维细胞高表达CTGF。缺氧可能是通过刺激成纤维细胞高表达CTGF,进而调节细胞分泌MMP-9的量,来达到促进细胞外基质沉积、气道重塑作用而形成纤维化。
Objective To observe the effects of different time of hypoxia on the expression of connective tissue growth factor (CTGF) and matrix metalloproteinase 9 (MMP-9) in lung fibroblasts. Methods Primary cultured human lung fibroblasts were incubated in hypoxia (37 ℃, 5% CO 2, 5% O 2, 90% N 2) for 0,1.5,3,6 and 12 h respectively. Reverse transcription polymerase chain reaction The expression of CTGF mRNA and MMP-9 mRNA were measured by RT-PCR. The concentrations of CTGF and MMP-9 in supernatant of culture supernatant were determined by enzyme-linked immunosorbent assay (ELISA). Results High expression of CTGF mRNA in hypoxia-stimulated fibroblasts occurred in 1.5h, and remained a relative plateau to 6h, and began to decline at 12h. MMP-9 mRNA was highly expressed at 1.5 h of hypoxia and maintained a continuously increasing trend. ELISA results confirmed that CTGF had the same peak concentration as MMP-9 at 12 h of hypoxia. Compared with the non-intervention group, the amount of MMP-9 in fibroblasts treated with anti-CTGF antibody was significantly decreased. Conclusion Hypoxia can promote high expression of CTGF in human lung fibroblasts. Hypoxia may be through the stimulation of fibroblasts high expression of CTGF, and then regulate the amount of MMP-9 cells to promote the promotion of extracellular matrix deposition, airway remodeling and the formation of fibrosis.