Anti-proliferative Capsular-like Polysaccharide Antigen from Actinobacillus actinomycetemcomitans Induces Apoptotic Cell Death in Mouse Osteoblastic MC3T3-E1 Cells
Anti-proliferative Capsular-like Polysaccharide Antigen from Actinobacillus actinomycetemcomitans Induces Apoptotic Cell Death in Mouse Osteoblastic MC3T3-E1 Cells
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DOI：
10.1177/00220345990780060601
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发表时间：
1999-06
影响因子：
7.6
通讯作者：
S. Yamamoto;Makio Mogi;Kyoko Kinpara;Y. Ishihara;N. Ueda;K. Amano;Tatsuji Nishihara;Toshihide Noguchi;A. Togari
中科院分区：
文献类型：
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作者：
S. Yamamoto;Makio Mogi;Kyoko Kinpara;Y. Ishihara;N. Ueda;K. Amano;Tatsuji Nishihara;Toshihide Noguchi;A. Togari
伴放线放线杆菌 (A. actinomycetemcomitans) 与局部幼年性牙周炎 (LJP) 的病因有关，并产生多种组织损伤产物。在这些产品中，来自 A. actinomycetemcomitans 的荚膜样多糖抗原 (CPA) 是骨吸收的有效介质。事实上，已知这种 CPA（b 血清型）可通过小鼠骨髓培养物中白细胞介素 (IL)-1α 的产生来促进破骨细胞样细胞的形成。尽管成骨细胞完成骨形成，但很少有报道关注CPA对炎症性疾病部位成骨细胞成骨活性的影响。我们假设 CPA 在成骨细胞中发挥介导作用。因此，本研究的目的是检测 A. actinomycetemcomitans 的 CPA 对小鼠成骨细胞系 MC3T3-E1 和人骨肉瘤 SaOS-2 细胞的影响。 A. actinomycetemcomitans 血清型 c 对两种细胞系的细胞增殖产生有效的剂量依赖性抑制。 CPA中抗增殖活性的表征表明它对MC3T3-E1没有细胞毒性。通过细胞 DNA 片段化 ELISA 和 FACS 分析评估，与 CPA-c 一起孵育 20 小时导致细胞凋亡细胞死亡显着增加。与使用细胞因子混合物（肿瘤坏死因子-α、IL-1β 和干扰素-γ）获得的结果相反，与 CPA-c 孵育后，在 MC3T3-E1 中未检测到诱导型一氧化氮 (NO) 合酶基因表达或 NO 释放。此外，CPA-b 和 -c 均能有效诱导细胞凋亡相关修饰物，例如 Fas mRNA，而 bcl-2 mRNA 水平没有变化。因此，本研究表明，A. actinomycetemcomitans 中的 CPA 含有一种有效的抗增殖多糖，其活性与 MC3T3-E1 中的细胞凋亡相关，并且 CPA 本身是 Fas 系统而非 NO 介导的细胞凋亡诱导剂。
Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) has been implicated in the etiology of localized juvenile periodontitis (LJP), and produces a multiplicity of tissue-damaging products. Among those products, the capsular-like polysaccharide antigen (CPA) from A. actinomycetemcomitans is a potent mediator of bone resorption. In fact, this CPA (serotype b) is known to promote osteoclast-like cell formation via interleukin (IL)-1α production in mouse marrow cultures. Although osteoblasts complete bone formation, there are few reports focusing on the effect of CPA in bone-forming activity of osteoblasts in inflammatory disease sites. We hypothesized that CPA plays a mediating role in osteoblastic cells. Therefore, the purpose of this study was to examine the effect of CPA from A. actinomycetemcomitans on the mouse osteoblastic cell line MC3T3-E1 and human osteosarcoma SaOS-2 cells. A. actinomycetemcomitans serotype c resulted in a potent dose-dependent inhibition of cell proliferation of both cell lines. Characterization of the antiproliferative activity in the CPA demonstrated that it was not cytotoxic for MC3T3-El. A 20-hour incubation with CPA-c resulted in a significant increase in apoptotic cell death in the cells, as evaluated by both cellular DNA fragmentation ELISA and FACS analysis. In contrast to the results obtained with a cytokine mixture (tumor necrosis factor-α, IL-1β, and interferon-γ), no inducible nitric oxide (NO) synthase gene expression or NO release could be detected in MC3T3-E1 after incubation with CPA-c. Further, both CPA-b and -c caused potent induction of apoptosis-related modifiers, e.g., Fas mRNA, whereas bcl-2 mRNA levels were unchanged. Therefore, this study has shown that CPA from A. actinomycetemcomitans contains a potent antiproliferative polysaccharide whose activity is associated with apoptotic cell death in MC3T3-E1, and that CPA per se is an inducer of apoptosis mediated by the Fas system but not by NO.