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目的探讨人巨细胞病毒(HCMV)感染对体外培养肺成纤维细胞(HEL)中明胶酶活性的影响。方法体外培养HEL细胞感染HCMV,分为低感染复数(MOI)组及高MOI组,每组重复6例。明胶酶谱法检测HEL细胞中MMP-2及MMP-9的明胶酶活性,用半定量RT-PCR检测各组HEL细胞中MMP-9及TIMP-1的转录水平。结果在低MOI组及高MOI组HEL细胞中MMP-9及MMP-2活性均增强(P<0.05),高MOI组MMP-9及MMP-2活性较低MOI组显著增加(P<0.05)。进一步检查MMP-9及TIMP-1的mRNA水平发现,正常对照组HEL细胞中MMP-9及TIMP-1的mRNA处于一个较低的水平,HCMV感染使HEL细胞中MMP-9及TIMP-1的mRNA水平均明显升高(P<0.05),低MOI组和高MOI组差异无统计学意义(P>0.05)。在低MOI组及高MOI组,HEL细胞MMP-9/TIMP-1的比值和正常对照组相比明显升高(P<0.05),表明MMP-9升高更为显著。高MOI组和低MOI组中MMP-9/TIMP-1的比值差异无统计学意义(P>0.05)。结论HCMV感染可以造成MMP-9和TIMP-1转录和MMP-9/TIMP-1的失衡,同时造成MMP-9及MMP-2明胶酶活性增强,导致肺泡结构的破坏和肺纤维化的发生,这在CMV肺炎的发病机制中起着重要的作用。
Objective To investigate the effect of human cytomegalovirus (HCMV) infection on gelatinase activity in cultured lung fibroblasts (HEL). Methods HEL cells were infected with HCMV in vitro and divided into low MOI group and high MOI group, with 6 replicates in each group. Gelatin zymography was used to detect the gelatinase activity of MMP-2 and MMP-9 in HEL cells. The transcription levels of MMP-9 and TIMP-1 in HEL cells were detected by semi-quantitative RT-PCR. Results The activities of MMP-9 and MMP-2 in HEL cells in low MOI group and high MOI group were significantly increased (P <0.05), while in high MOI group, the activities of MMP-9 and MMP-2 were significantly increased (P <0.05) . Further examination of mRNA levels of MMP-9 and TIMP-1 found that mRNA of MMP-9 and TIMP-1 in HEL cells of normal control group was at a low level. HCMV infection caused MMP-9 and TIMP-1 mRNA levels were significantly increased (P <0.05), but there was no significant difference between low MOI group and high MOI group (P> 0.05). In low MOI group and high MOI group, the ratio of MMP-9 / TIMP-1 in HEL cells was significantly higher than that in the normal control group (P <0.05), indicating that MMP-9 increased more significantly. The ratio of MMP-9 / TIMP-1 in high MOI group and low MOI group had no significant difference (P> 0.05). Conclusion HCMV infection can cause the imbalance of MMP-9 and TIMP-1 transcription and MMP-9 / TIMP-1, and increase the activity of MMP-9 and MMP-2 gelatinase, leading to the destruction of alveolar structure and pulmonary fibrosis. This plays an important role in the pathogenesis of CMV pneumonia.