Experimental pathological study on roles of chemokines in LPS-induced periodontal tissue destruction - Basic analysis on remedical effects of specific antibody against chemokine -

1. 局部应用脂多糖（LPS）后，巨噬细胞炎症蛋白-2 （MIP-2）和巨噬细胞化学引诱蛋白-1 （mcp -1）在大鼠牙周组织中的免疫表达：1)从大鼠牙龈沟局部应用5 mg/ml大肠杆菌脂多糖，可迅速引起大鼠牙周结上皮（JE）和亚JE区炎症变化。此外，观察到沿牙槽骨表面的破骨细胞骨吸收刺激在3小时和3天出现双相反应高峰。2)观察到MIP-2在乙脑细胞中的瞬时免疫表达在第1天达到峰值。与MIP-2的过度表达相对应，LPS可诱导乙脑和亚乙脑区中性粒细胞数量显著增加。3)应用LPS后第1天和第2天，观察肺泡嵴区成骨细胞和骨细胞中MCP-1的免疫表达。这些细胞产生的MCP-1可能会引起局部破骨前细胞的募集和随后的破骨细胞的聚集。不同浓度MIP-2的应用：不同浓度的重组大鼠MIP-2（0.05、0.5、5、50 μg/ml）从龈沟处应用，不仅能引起中性粒细胞向乙脑和亚乙脑区募集，还能引起破骨细胞沿牙槽骨表面呈剂量依赖性聚集3。先前腹腔注射抗mip -2抗体对LPS应用引起的牙周组织破坏的影响：注射抗mip -2抗体可显著减少中性粒细胞向乙脑和乙脑亚区募集，并在LPS应用后3天显著减少破骨细胞的出现。另一方面，破骨细胞在3小时的第一次增加，似乎是LPS的直接作用，不受抗mip -2抗体处理的抑制。这些研究结果表明MIP-2可能是一种强大的cxc趋化因子，用于中性粒细胞募集到牙周病部位，并可能在炎症的发生中起重要作用。此外，牙周病部位过度分泌cxc趋化因子可能诱导各种破骨细胞刺激因子的产生，导致随后的牙槽骨破坏。提示建立以cxc趋化因子为靶点的牙周治疗新方法的可能性。少

1. Immunoexpression of macrophage inflammatory protein-2 (MIP-2) and macrophage chemoattractant protein-1 (MCP-l) in rat periodontal tissue after topical application of Lipopolysaccharide (LPS) : 1) Topically applied 5 mg/ml E. Coli LPS from rat gingival sulcus rapidly provoked inflammatory changes in junctional epithelium (JE) and sub-JE area. In addition, stimulation of osteoclastic bone resorption along the alveolar bone surface showing a biphasic response peaking at 3 hours and 3 days was observed. 2) A transient immuno-expression of MIP-2 in JE cells peaking at 1 day was observed. Corresponding to excessive MIP-2 expression, LPS application induced a significant increase in number of neutrophils in JE and sub-JE area. 3) At day 1 and 2 after LPS application, immunoexpression of MCP-1 in osteoblasts and osteocytes presented in alveolar crestal area was observed. The MCP-1 production from these cells may cause local recruitment of preosteoclasts and the following osteoclastic bone r … More esorption at day 3.2. Application of various concentrations of MIP-2 : Various concentrations of recombinant rat MIP-2 (0.05, 0.5, 5, 50 μg/ml) applied from gingival sulcus caused not only neutrophil recruitment into JE and sub-JE area but also osteoclasts amassment along the alveolar bone surface in a dose dependent manner3. Effects of previously intraperitoneal-injected anti-MIP-2 antibody on periodontal tissue destruction caused by LPS-application : Injection of the anti-MIP-2 antibody remarkably reduced the recruitment of neutrophils into JE and sub-JE area and significantly decreased the appearance of osteoclasts at 3 days after LPS application. On the other hand, the first increase of osteoclasts at 3 hours, which is seemed to be a direct effect of LPS, was not inhibited by the anti-MIP-2 antibody treatment. Theses findings indicated that MIP-2 may be one of powerful CXC-chemokines for neutrophil recruitment into the periodontal disease site and may play an important role in the initiation of inflammation. Moreover, the over secretion of CXC-chemokine in the periodontal disease site may induce production of various osteoclast stimulating factors resulting in the subsequent alveolar bone destruction. It is suggested a possibility of establishment new periodontal therapy targeting CXC-chemokine. Less