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

• 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

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

内毒素在牙周病牙槽骨破骨细胞吸收中起重要作用。虽然内毒素在体内和体外都能刺激破骨细胞性骨吸收，但其机制尚不清楚。最近我们发现人脐血和骨髓细胞培养物可诱导形成表达破骨细胞的多核细胞，并提出CFU-GM(集落形成单位-粒细胞-巨噬细胞)是破骨细胞的前体细胞。本实验研究了牙龈假单胞菌和大肠杆菌的胞外产物脂多糖对CFU-GM来源的细胞形成破骨细胞和骨吸收的生物学作用。在这些培养物中加入10-100 ng/ml的脂多糖(10-100 ng/ml)，可显著增加23C6阳性…的形成。加入抗人IL-I可完全抑制任一种LPS刺激的单核细胞形成。更多的PS刺激骨吸收，但牙龈卟啉单胞菌(Pgigivaris)脂多糖引起的吸收面积增加是大肠杆菌(E.coli)的1.4倍。在破骨细胞前体细胞的实验中，还研究了内毒素对酪氨酸磷酸化的调节作用。在脂多糖处理的CFU-GM细胞和多核细胞中可检测到磷酸化。此外，在每个细胞的边缘也可检测到磷化。当这些细胞与牙龈假单胞菌的脂多糖孵育时，检测到一条42kD的含有磷酸酪氨酸的蛋白条带。加入100 nM的除草霉素A可完全抑制细菌脂多糖和大肠杆菌脂多糖刺激的骨吸收，但不能抑制单个核细胞的形成。这些实验表明，脂多糖通过对破骨细胞前体细胞的初级作用刺激破骨细胞的形成，破骨细胞前体细胞是由内毒素产生的IL-1作用而诱导的，IL-1反过来刺激破骨细胞的形成，导致破骨细胞性骨吸收。这些发现还表明，c-src激酶参与了脂多糖对破骨细胞骨吸收活性的调节。较少

项目成果

下山雅通,辰巳順一,栗原徳善: "歯周病原性細菌のリポ多糖体による破骨細胞形成機構の解析" 日本歯周病学会会誌. 39. 313-323 (1997)

Masamichi Shimoyama、Junichi Tatsumi、Tokuyoshi Kurihara：“牙周病原菌脂多糖的破骨细胞形成机制分析”日本牙周病学会杂志 39. 313-323 (1997)。