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目的观察支原体巨噬细胞活化脂肽-2(MALP-2)对人单核细胞分泌基质金属蛋白酶-9(MMP-9)的影响。方法体外培养人单核细胞系THP-1,分别采用0、0.1、1.0和5.0μg/m L MALP-2刺激THP-1细胞12~18 h,采用实施定量PCR检测MMP-9 m RNA的表达,荧光共振能量转移(FRET)检测MMP-9的酶活性变化,ELISA检测培养上清中MMP-9的含量。结果 THP-1细胞未刺激时,MMP-9的m RNA表达量、酶活性以及分泌水平极低。当给予0.1~5.0μg/m L MALP-2作用18 h后,MMP-9的分泌水平显著增高,细胞培养上清液中MMP-9的酶活性也明显增强。此外,5.0μg/m L MALP-2刺激THP-1细胞6 h后即可诱导MMP-9 m RNA表达,16 h后达到峰值。结论 MALP-2可诱导人单核细胞表达MMP-9,这可能是支原体感染早期重要的致病因素。
Objective To investigate the effect of MALP-2 on the secretion of matrix metalloproteinase-9 (MMP-9) from human monocytes. Methods Human monocytic THP-1 cells were cultured in vitro and THP-1 cells were stimulated with 0, 0.1, 1.0 and 5.0 μg / mL MALP-2 for 12-18 h respectively. The expression of MMP-9 mRNA was detected by real- The changes of MMP-9 activity were detected by fluorescence resonance energy transfer (FRET). The content of MMP-9 in the culture supernatant was detected by ELISA. Results When THP-1 cells were not stimulated, the expression of m RNA, the activity of MMP-9 and the secretion level of MMP-9 were very low. When MALP-2 was treated with 0.1 ~ 5.0 μg / mL for 18 h, the secretion of MMP-9 was significantly increased, and the activity of MMP-9 in the cell culture supernatant was significantly increased. In addition, MMP-9 mRNA expression was induced by 5.0μg / mL MALP-2 stimulated THP-1 cells for 6 h and peaked at 16 h. Conclusion MALP-2 can induce the expression of MMP-9 in human monocytes, which may be an important virulence factor in the early stage of mycoplasma infection.