Chang Jae-Ho, Kim Soo-Ki, Choi In-Hong, Lee Sang-Kyou, Morio Tomohiro, Chang Eun-Ju
Institute of Basic Medical Science, Wonju College of Medicine, Yonsei University, Wonju 220-701, Republic of Korea.
Int J Biochem Cell Biol. 2006;38(4):638-47. doi: 10.1016/j.biocel.2005.11.005. Epub 2005 Dec 5.
Trichomonas vaginalis, a flagellated protozoan parasite, is the causative organism of trichomoniasis. We have recently demonstrated that T. vaginalis induces apoptotic cell death via a Bcl-x(L)-dependent pathway in RAW264.7 macrophages. In this study, we attempted to characterize in detail the signaling cascades resulting in T. vaginalis-induced macrophage apoptosis, focusing particularly on mitochondrial changes and the role of p38 mitogen-activated protein kinase (p38 MAPK) activation. We found that T. vaginalis induced mitochondrial changes including the release of cytochrome c and the serial activation of caspases, leading to the activation of p38 MAPK in macrophages. These biochemical changes culminated in the apoptosis of the host cells. Caspase inhibitors induced a significant inhibition of T. vaginalis-induced nuclear damage, as well as the activation of p38 MAPK. Treatment with the p38 MAPK inhibitor, SB203580, or the overexpression of kinase-inactive p38 MAPK, induced an attenuation of T. vaginalis-induced apoptosis but not cytochrome c release, the activation of caspase-9 and caspase-3, or PARP cleavage. Furthermore, SB203580 treatment to human macrophages consistently blocked T. vaginalis-induced apoptosis. Collectively, our findings indicate that p38 MAPK signaling cascade is requisite to apoptosis of T. vaginalis-infected macrophage, and this apoptotic process occurs via the phosphorylation of p38 MAPK, which is located downstream of mitochondria-dependent caspase activation, conferring insight into the plausible molecular mechanism of T. vaginalis-immune evasion from macrophage attack.
阴道毛滴虫是一种有鞭毛的原生动物寄生虫,是滴虫病的致病生物体。我们最近证明,阴道毛滴虫通过RAW264.7巨噬细胞中依赖Bcl-x(L)的途径诱导凋亡性细胞死亡。在本研究中,我们试图详细描述导致阴道毛滴虫诱导巨噬细胞凋亡的信号级联反应,特别关注线粒体变化以及p38丝裂原活化蛋白激酶(p38 MAPK)激活的作用。我们发现阴道毛滴虫诱导线粒体变化,包括细胞色素c的释放和半胱天冬酶的系列激活,导致巨噬细胞中p38 MAPK的激活。这些生化变化最终导致宿主细胞凋亡。半胱天冬酶抑制剂显著抑制阴道毛滴虫诱导的核损伤以及p38 MAPK的激活。用p38 MAPK抑制剂SB203580处理或激酶失活的p38 MAPK的过表达诱导阴道毛滴虫诱导的凋亡减弱,但不影响细胞色素c的释放、半胱天冬酶-9和半胱天冬酶-3的激活或PARP裂解。此外,用SB203580处理人巨噬细胞持续阻断阴道毛滴虫诱导的凋亡。总的来说,我们的研究结果表明,p38 MAPK信号级联反应是阴道毛滴虫感染巨噬细胞凋亡所必需的,并且这种凋亡过程通过p38 MAPK的磷酸化发生,p38 MAPK位于线粒体依赖性半胱天冬酶激活的下游,这为阴道毛滴虫逃避巨噬细胞攻击的可能分子机制提供了见解。