The study of signal transduction system on tyrosin phospholiration in LPS stumulated osteoclast-like cells

LPS刺激破骨细胞样细胞酪氨酸磷酸化信号转导系统的研究

基本信息

批准号：
09671963
负责人：
TATSUMI Junichi
金额：
$ 0.7万
依托单位：
Meikai University
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-09671963/
关键词：
Lipopolysaccharide Osteoclast Bone resorption Tyrosine phosphorylation c-src リポ多糖化 P.gingivalis

项目摘要

Lipopolysaccharide (LPS) plays important roles on osteoclastic resorption of alveolar bone in periodontal disease. Although LPS stimulates osteoclastic bone resorption in vivo and in vitro, but its mechanism is not clear. Recently we showed that human umbilical cord blood and bone marrow cell cultures can adapted to form multinucleate cells (MNC's) that express an osteoclast, and we also suggested that the CFU-GM (colony forming unit-granulocyte macrophage) is the progenitor for the osteoclast In the present experiments, we studied the biological effect of LPS, an extracellular product from P gingivalis and E.coli , on the mechanism of osteoclast formation from CFU-GM-derived cells and on bone resorption. Addition of LPS (10-100 ng/ml) from either of P gingivalis and E.co/i to these cultures significantly increased the formation of 23C6-positive MINCs.Addition of anti-human IL-i to cultures treated with LPS totally inhibited the increase in MNC formation stimulated by either LPS.Both L … More PS's stimulated bone resorption, but the Porphyromonas gingivalis (Pgingivaris) LPS caused a 1.4 fold greater increase in the resorption area compared with the Escherihia coli (E.coli) one. The effects of LPS on regulation of tyrosine phosphorylation were also studied in experiments utilizing osteoclast precursor cells. The phosphorylation was detectable in LPS-treated CFU-GM cells and multinucleated cells. Also the phospholiration was detectable in border of each cells. When these cells were incubated with LPS from P gingivalis, a 42-kD protein band containing phoshotyrosine was detected. Addition of 100 nM herbimycin A to cultures treated with either LPS totally inhibited the bone resorption stimulated by Pgingivalis and K coli LPS's. However, herbimycin A did not inhibit the MNC formation. These experiments suggest that LPS stimulates osteoclast formation through a primary action on osteoclast precursor cells, which cells are induced by the action of IL- 1 produced by the LPS which in turn stimulates osteoclast formation, resulting in osteoclastic bone resorption. These findings also suggest that c-src kinase is involved in the regulation of bone-resorbing activity of osteoclasts by LPS. Less

脂多糖（LPS）在牙周疾病中肺泡骨的整骨分辨率上起着重要作用。尽管LPS在体内和体外刺激了破骨骨骨分辨率，但其机制尚不清楚。 Recently we showed that human umbilical cord blood and bone marrow cell cultures can adapt to form multinucleate cells (MNC's) that express an osteoclast, and we also suggested that the CFU-GM (colony forming unit-granulocyte macrophage) is the progenitor for the osteoclast In the present experiments, we studied the biologic effect of LPS, an extracellular product from P牙龈和大肠杆菌，关于CFU-GM衍生细胞和骨骼分辨率的破骨细胞形成的机理。从p牙龈和e.co/i中添加LPS（10-100 ng/ml）到这些培养物，显着增加了23c6阳性Minc的形成。对用LPS治疗的抗人IL-I的添加完全抑制了MNC形成的MNC形成的增加。与大肠杆菌（E.Coli）相比，LPS的分辨率面积增加了1.4倍。 LPS对使用破骨细胞前体细胞的实验中还介绍了LPS对酪氨酸磷酸化调节的影响。在LPS处理的CFU-GM细胞和多核细胞中检测到磷酸化。同样在每个细胞的边界中检测到磷脂。当将这些细胞与P牙龈牙龈孵育时，检测到一个含有Phoshotyrosine的42 kD蛋白带。在用任何一种LPS治疗的培养物中添加100 nm的Herbimycin a完全抑制了pgingivalis和K Coli LPS刺激的骨骼分辨率。但是，除草霉素A没有抑制MNC的形成。这些实验表明，LPS通过对破骨细胞前体细胞的主要作用刺激破骨细胞的形成，该细胞是由LPS产生的IL-1的作用诱导的，从而刺激了破骨细胞的形成，从而导致破骨细胞的修复。这些发现还表明，C-SRC激酶参与LPS对破骨细胞的骨呈现活性的调节。较少的