Nonaka K, Ishisaki A, Muro M, Kato S, Oido M, Nakashima K, Kowashi Y, Nishihara T
Department of Oral Science, National Institute of Infectious Diseases, Tokyo, Japan.
FEMS Microbiol Lett. 1998 Feb 15;159(2):247-54. doi: 10.1111/j.1574-6968.1998.tb12868.x.
We have previously reported the evidence for apoptosis in the mouse macrophage cell line J774.1 by the periodontopathic bacterium Actinobacillus actinomycetemcomitans. In this study, we examined the role of protein kinases in the induction of apoptosis in A. actinomycetemcomitans-infected J774.1 cells by the MTT assay, fluorescence microscopy and flow cytometric analysis. After J774.1 cells were precultured with protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate (PMA), J774.1 cells infected with A. actinomycetemcomitans showed the increased percentage of apoptotic cells. On the contrary, protein kinase A (PKA) activators, such as forskolin and dibutyryl cAMP, do not mimic the effect of PMA. PKC inhibitors, such as staurosporine, calphostin C, chelerythrine chloride, and H7 were found to suppress apoptotic cell death in J774.1 cells infected with A. actinomycetemcomitans. However, HA1004, known as PKA inhibitor, had no effect on apoptosis in infected macrophages. The results presented here suggest that the signals through PKC may play crucial roles in the modulation of apoptosis in macrophages infected with A. actinomycetemcomitans.
我们之前曾报道过牙周病原菌伴放线放线杆菌可诱导小鼠巨噬细胞系J774.1发生凋亡的证据。在本研究中，我们通过MTT法、荧光显微镜检查和流式细胞术分析，研究了蛋白激酶在伴放线放线杆菌感染的J774.1细胞凋亡诱导中的作用。在用蛋白激酶C（PKC）激活剂佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯（PMA）预培养J774.1细胞后，感染伴放线放线杆菌的J774.1细胞凋亡细胞百分比增加。相反，蛋白激酶A（PKA）激活剂，如福斯高林和二丁酰环磷腺苷，不能模拟PMA的作用。发现PKC抑制剂，如星形孢菌素、钙泊三醇、氯化白屈菜红碱和H7，可抑制感染伴放线放线杆菌的J774.1细胞中的凋亡细胞死亡。然而，作为PKA抑制剂的HA1004对感染巨噬细胞的凋亡没有影响。此处给出的结果表明，通过PKC的信号可能在伴放线放线杆菌感染的巨噬细胞凋亡调节中起关键作用。
