Zeng Yanhua, Wu Yimou, Yu Minjun, Liu Jie, You Xiaoxing, Tang Shuangyang
Institute of Pathogenic Biology, Medical College, University of South China, Hengyang 421001, China.
Wei Sheng Wu Xue Bao. 2008 Mar;48(3):355-61.
To investigate the potential pathogenicity of Mycoplasma penetrans (M. penetrans) and the possible molecular mechanisms, we investigated whether M. penetrans lipid-associated membrane proteins (LAMPs) could induce mouse macrophages Raw264.7 apoptosis by activating nuclear factor kappaB (NF-kappaB). Apoptosis was detected in M. penetrans LAMPs-stimulated mouse macrophages by Annexin-V-FITC staining and DNA fragmentation analysis. We also analyzed the activation of NF-kappaB and the effects of pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kappaB, on M. penetrans LAMPs-induced mouse macrophages apoptosis by indirect immunofluorescence and Western blotting. Our results suggested that M. penetrans LAMPs could induce mouse macrophages significant early- and late-stage apoptosis. Agarose gel electrophoresis of the DNA of LAMPs-challenged cells revealed that a ladder-like pattern of migration of DNA indicative of apoptosis. M. penetrans LAMPs could activate NF-kappaB in mouse macrophages. PDTC could partially inhibit the activation of NF-kappaB and thus inhibit M. penetrans LAMPs-induced mouse macrophages apoptosis. This study demonstrates that M. penetrans LAMPs may be an important etiological factor due to its ability to induce mouse macrophages apoptosis, which was probably mediated through the activation of NF-kappaB.
为了研究穿透支原体(M. penetrans)的潜在致病性及其可能的分子机制,我们研究了穿透支原体脂相关膜蛋白(LAMPs)是否能通过激活核因子κB(NF-κB)诱导小鼠巨噬细胞Raw264.7凋亡。通过Annexin-V-FITC染色和DNA片段化分析检测穿透支原体LAMPs刺激的小鼠巨噬细胞中的凋亡情况。我们还通过间接免疫荧光和蛋白质印迹分析了NF-κB的激活情况以及NF-κB抑制剂吡咯烷二硫代氨基甲酸盐(PDTC)对穿透支原体LAMPs诱导的小鼠巨噬细胞凋亡的影响。我们的结果表明,穿透支原体LAMPs可诱导小鼠巨噬细胞发生明显的早期和晚期凋亡。对LAMPs刺激细胞的DNA进行琼脂糖凝胶电泳显示出指示凋亡的DNA梯形迁移模式。穿透支原体LAMPs可激活小鼠巨噬细胞中的NF-κB。PDTC可部分抑制NF-κB的激活,从而抑制穿透支原体LAMPs诱导的小鼠巨噬细胞凋亡。本研究表明,穿透支原体LAMPs可能是一个重要的致病因素,因为它能够诱导小鼠巨噬细胞凋亡,这可能是通过激活NF-κB介导的。