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Interleukin-6 stimulates cathepsin B and L activities of human periodontal ligament cells through the signaling pathways

Interleukin-6通过信号通路刺激人牙周膜细胞的组织蛋白酶B和L活性

基本信息

批准号：
14571969
负责人：
YAMAGUCHI Masaru
金额：
$ 2.18万
依托单位：
Nihon University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571969/
关键词：
Human periodontal ligament cells Tension force Compression force Gingival crevicular fluid Cathepsin B, L Experimental tooth movement Interleukin-6 培養ヒト歯根膜線維芽細胞カテプシンB、L 矯正学的歯の移動カテプンB, L

项目摘要

Our in vitro study examined the secretion of cathepsin B(CAB) and cathepsin L(CAL) in PDL cells following mechanical stress. PDL cells were subjected to 0.5, 1.0, 2.0, or 3.0 g/cm^2 of compression force or an increase in surface area by tension force of 0.28%, 0.95%, 1.72%, or 2.50% for 24 hr. We found compression and tension significantly increased the secretions of both CAB and CAL in PDL cells, which were exhibited in a time- and force magnitude-dependent manner. The compression-stimulated secretion of CAB was increased approximately 3-fold and that of CAL 4-fold, as compared with the control. Further, tension-stimulated secretion of CAB was increased by 1.5-fold and that of CAL 2-fold compared with the control. When analyzed using a semiquantitative polymerase chain reaction(RT-PCR) assay, CAB and CAL mRNA were increased in response to both compression and tension forces. These findings demonstrated that mechanical stress (compression and tension forces) causes an increase in secretion of CAB and CAL in PDL cells in vitro.Furthermore, we examined that the mechanisms controlling CAB and CAL activities and intracellular signaling by IL-6 in orthodontic tooth movement. Cultured PDL cells were treated with different concentrations (0.01-10 ng/ml) of IL-6 in the absence or presence of the p38 MAP kinase inhibitor SB-208350 or the p42/44 inhibitor PD-98059. Protein and gene expression of CAB and CAL were determined by ELISA and PCR, respectively. SB-208350 and PD-98059 inhibited the increase in CAB and CAL production induced by IL-6. These results suggest the IL-6 regulates muscle CAB and CAL production at least in part by activating the MAP kinase pathway and NF-kappaB.

我们在体外研究了组织蛋白酶B（CA B）和组织蛋白酶L（CAL）在PDL细胞中的分泌后，机械应力。将PDL细胞置于0.5、1.0、2.0或3.0 g/cm ^2的压力或0.28%、0.95%、1.72%或2.50%的张力下24小时，我们发现压力和张力显著增加PDL细胞中CAB和CAL的分泌，其以时间和力大小依赖的方式显示。与对照组相比，压缩刺激的CAB分泌增加约3倍，CAL分泌增加约4倍。此外，与对照组相比，张力刺激的CAB分泌增加了1.5倍，CAL分泌增加了2倍。当使用半定量聚合酶链反应（RT-PCR）分析时，CAB和CAL mRNA的增加，在响应压缩和张力。这些结果表明，机械应力（压力和张力）导致牙周膜细胞分泌的CAB和CAL的增加，在体外，我们进一步研究了控制CAB和CAL的活性和细胞内信号转导的IL-6在正畸牙齿移动的机制。在不存在或存在p38 MAP激酶抑制剂SB-208350或p42/44抑制剂PD-98059的情况下，用不同浓度（0.01-10 ng/ml）的IL-6处理培养的PDL细胞。ELISA和PCR分别检测CAB和CAL的蛋白和基因表达。SB-208350和PD-98059抑制IL-6诱导的CAB和CAL产生的增加。这些结果表明IL-6至少部分通过激活MAP激酶途径和NF-κ B来调节肌肉CAB和CAL的产生。