Role of IL-beta converting enzyme on the induction of apoptosis mediated by the infection of periodontopathic bacteria infection

IL-β转换酶在牙周病菌感染介导的细胞凋亡诱导中的作用

• 批准号: 09671885
• 负责人: NISHIHARA Tatsuji
• 金额: $ 1.86万
• 依托单位: National Institute of Infectious Diseases
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671885/
• 关键词: periodontopathic bacteria; periodontitis; apoptosis; interleukin-1beta; caspase; 歯周病; 細菌感染; 歯周病細菌; IL-1β; IL-1β変換酵素; IL-1β変換酵素阻害剤; マクロファージ

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. After J774.1 cells were precultured with protein kinase C (PKC) activator, J774.1 cells infected with A.actinomycetemcomitans showed the increased percentage of apoptotic cells. On the contrary, PKA activator do not mimic the effect of PKC activator. In addition, PKC inhibitors suppressed apoptotic cell death in J774.1 cells infected with A.actinomycetemcomitans. 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. Next, we examined the role of caspases, interleukin-1beta (IL-1beta) converting enzyme family proteases, in the regulation of apoptosis in infected J774.1 cells. TWo peptide inhibitors of caspases, benzyloxycarbonyl-Val-Ala-Asp (OMe)-fluoromethyl ketone (Z-VAD-FMK ; caspase-1 inhibitor)and benzyloxycarbonyl-Asp-Glu-Val-ASP (OMe)-fluoromethyl ketone (DEVD-FMK ; caspase-3 inhibitor) inhibited the induction of apoptosis in J774.1 cells infected with A.actinomycetemcomitans. Furhtermore, Z-VAD-FMK suppressed the production of IL-1beta from A.actinomycetemcomitans-infected J774.1 cells. When endogenous caspase-3 activity of J774.1 cells were analyzed, caspase-3 activity was markedly increased after A.actinomycetemcomitans infection. In addition, Z-VAD-FMK and DEVD-FMK suppressed caspase-3 activity of J774.1 cells infected with A.actinomycetemcomitans. These findings suggest that caspase-1 and-3 are involved in the regulation of apoptosis in macrophages infected with A.actinomycetemcomitans.

我们以前曾报道过牙周病细菌伴放线放线杆菌引起小鼠巨噬细胞系J774.1凋亡的证据。在这项研究中，我们研究了蛋白激酶在诱导细胞凋亡中的作用，放线菌共生放线杆菌感染的J774.1细胞。用蛋白激酶C（PKC）激活剂预培养J774.1细胞后，感染伴放线菌的J774.1细胞凋亡率增加。相反，PKA激活剂不能模拟PKC激活剂的作用。此外，PKC抑制剂抑制细胞凋亡的J774.1细胞感染放线菌共生放线菌。这里提出的结果表明，通过PKC的信号可能发挥至关重要的作用，在巨噬细胞感染放线菌伴放线菌的细胞凋亡的调制。接下来，我们研究了半胱天冬酶，白细胞介素-1 β（IL-1 β）转换酶家族蛋白酶，在受感染的J774.1细胞凋亡的调节中的作用。半胱天冬酶的两种肽抑制剂，苄氧羰基-Val-Ala-Asp（OMe）-氟甲基酮（Z-VAD-FMK ;半胱天冬酶-1抑制剂）和苄氧羰基-Asp-Glu-Val-ASP（OMe）-氟甲基酮（DEVD-FMK ;半胱天冬酶-3抑制剂）抑制伴放线放线菌感染的J774.1细胞的凋亡诱导。此外，Z-VAD-FMK还能抑制伴放线杆菌感染的J774.1细胞产生IL-1 β。当分析J774.1细胞的内源性caspase-3活性时，伴放线菌感染后caspase-3活性显著增加。此外，Z-VAD-FMK和DEVD-FMK抑制caspase-3活性的J774.1细胞感染放线菌共生放线菌。这些研究结果表明，caspase-1和-3参与调节细胞凋亡的巨噬细胞感染放线菌伴放线菌。

项目成果

Hyashida,H.et al.: "The differentiation of clinical isolates of Actinobacillus actinomycetemconitans by using a insertion sequence ISAa1" Oral Microbiol.Immunol.13. 120-123 (1998)

Hyashida,H.et al.：“通过使用插入序列 ISAa1 来区分 Actinobacillus actinomycetemconitans 的临床分离株”Oral Microbiol.Immunol.13。

Nonaka,K.et al.: "Possible involvement of protein KinaseC in apoptotic cell death of macrophages infected with Actinobacillus actinanycetemconitars" FEMS Microbiol.Lett.159. 247-254 (1998)

Nonaka,K.et al.：“蛋白激酶 C 可能参与感染 Actinobacillus actinanycetemconitars 的巨噬细胞凋亡细胞死亡”FEMS Microbiol.Lett.159。

Ueda,N.et al.: "Involvement of prostaglandin E2 and interleukin-1α in the differentiation and survival of osteoclast induced by lipopolysaccharide from Actinobacillus actinomycetemcomtans Y4" J. Periodont. Res.33. 509-516 (1998)

Ueda, N. 等人：“前列腺素 E2 和白细胞介素 1α 在由 Actinobacillus actinomycetemcomtans Y4 脂多糖诱导的破骨细胞分化和存活中的参与”J. periodont. 509-516 (1998)。

Hayashida,H.,et al.: "The differentiation of clinical isolates of Actinobacillus actinomycetemconitaus by using a insertion sequence ISAa1" Oral Microbiology Immunology. (印刷中). (1998)

Hayashida, H., et al.：“通过使用插入序列 ISAa1 区分放线杆菌的临床分离株”口腔微生物学免疫学（1998 年出版）。

Ueda,N.et al.: "Involvement of prostagland in E_2 and interleukin-1α in the differentration and survival of osteoclasts induced by lipopolysaccharide from Actinobacillus actinomycetemcontars Y4" J.Periodont.Res.33. 509-516 (1998)

Ueda，N.等人：“前列腺在E_2和白介素-1α中参与由放线杆菌放线菌Y4的脂多糖诱导的破骨细胞的分化和存活”J.Periodont.Res.33(1998)。