Qiu Hong, Wu Yi-Mou, You Xiao-Xing, Yu Ming-Jun, Zeng Yan-Hua
Department of Microbiology, Medical College, Nanhua University, Hengyang 421001, China.
Wei Sheng Wu Xue Bao. 2007 Oct;47(5):899-904.
Designed to investigate the potential pathogenicity of Mycoplasma genitalium (M. genitalium) and its molecular mechanisms responsible for the induction of proinflammatory cytokines gene expression in human monocytic cells (THP-1) stimulated by lipid-associated membrane proteins (LAMPs) prepared from M. genitalium. THP-1 cells were stimulated with LAMPs to analyze the production of proinflammatory cytokines and the expression of mRNA was detected by RT-PCR. Cell apoptosis was detected in THP-1 cells by Annexin V-propidium iodide staining. The activity of transcriptional factors, NF-kappaB, was examined in THP-1 cells treated with LAMPs by EMSA. The effects of pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kappaB, on the production of proinflammatory cytokines, the expression of mRNA and apoptosis were also examined in THP-1 cells treated with LAMPs. M. genitalium LAMPs stimulate THP-1 cells to produce TNF-alpha, IL-1beta and IL-6 in dose- and time-dependent manner. The mRNA levels and cell apoptosis are also downregulated in response to LAMPs stimulation and inhibited by PDTC treatment. M. genitalium LAMPs are found to trigger NF-kappaB activation, a possible mechanism for the induction of mRNA expression and the cell apoptosis. This study demonstrated that M. genitalium may be an important etiological factor of certain disease due to the ability of LAMPs to stimulated the expression of mRNA and apoptosis, which is probably mediated through the activation of NF-kappaB.
旨在研究生殖支原体(M. genitalium)的潜在致病性及其在由生殖支原体制备的脂质相关膜蛋白(LAMPs)刺激的人单核细胞(THP-1)中诱导促炎细胞因子基因表达的分子机制。用LAMPs刺激THP-1细胞以分析促炎细胞因子的产生,并通过RT-PCR检测mRNA的表达。通过Annexin V-碘化丙啶染色检测THP-1细胞中的细胞凋亡。通过EMSA检测在用LAMPs处理的THP-1细胞中转录因子NF-κB的活性。还在用LAMPs处理的THP-1细胞中检测了NF-κB抑制剂吡咯烷二硫代氨基甲酸盐(PDTC)对促炎细胞因子产生、mRNA表达和细胞凋亡的影响。生殖支原体LAMPs以剂量和时间依赖性方式刺激THP-1细胞产生TNF-α、IL-1β和IL-6。响应LAMPs刺激,mRNA水平和细胞凋亡也下调,并被PDTC处理抑制。发现生殖支原体LAMPs触发NF-κB活化,这是诱导mRNA表达和细胞凋亡的一种可能机制。这项研究表明,由于LAMPs刺激mRNA表达和细胞凋亡的能力,生殖支原体可能是某些疾病的重要病因,这可能是通过NF-κB的激活介导。