Oyama T, Matsushita K, Sakuta T, Tokuda M, Tatsuyama S, Nagaoka S, Torii M
Department of Restorative Dentistry and Endodontology, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan.
J Periodontal Res. 2007 Feb;42(1):53-61. doi: 10.1111/j.1600-0765.2006.00914.x.
In periodontitis, matrix metalloproteinases (MMPs) are upregulated in response to locally released inflammatory cytokines, resulting in pathologic processes. Roxithromycin is a 14-membered ring macrolide antibiotic with broad-spectrum antibacterial effects against oral pathogens and immunomodulatory effects. Recently, we reported that roxithromycin inhibits tumor necrosis factor (TNF)-alpha-induced vascular endothelial growth factor expression in human periodontal ligament (HPDL) cell cultures. In the present study, we examined the effect of roxithromycin on TNF-alpha-induced MMP-1 production by HPDL cells.
Cultured cells were incubated with 1% fetal bovine serum for 24 h, followed by treatment with 10 ng/ml TNF-alpha, 10 microM roxithromycin, and mitogen-activated protein kinase inhibitor at various concentrations. Culture supernatants and sediments were collected at different time-points and used for enzyme-linked immunosorbent assays, and northern and western blot analyses.
In HPDL cell cultures, roxithromycin strongly inhibited TNF-alpha-induced MMP-1 mRNA expression and production. The inhibition of MMP-1 gene expression by roxithromycin was dependent on de novo protein synthesis and was regulated at the transcriptional level. Roxithromycin significantly inhibited TNF-alpha-induced c-Jun N-terminal kinase activation (JNP) and marginally inhibited extracellular signal-regulated kinase (ERK) 1/2 activation, but not p38 mitogen-activated protein kinase activation. Furthermore, roxithromycin reduced the induction of Ets-1, one of the critical factors in MMP-1 transcription.
Roxithromycin inhibits TNF-alpha-mediated MMP-1 induction through the downregulation of ERK1/2 and JNK activation and the subsequent reduction of Ets-1, suggesting that roxithromycin may have therapeutic use in periodontitis and other chronic inflammatory conditions involving MMP-1 induction.
在牙周炎中，基质金属蛋白酶（MMPs）会因局部释放的炎性细胞因子而上调，从而引发病理过程。罗红霉素是一种14元环大环内酯类抗生素，对口腔病原体具有广谱抗菌作用且有免疫调节作用。最近，我们报道罗红霉素在人牙周膜（HPDL）细胞培养中可抑制肿瘤坏死因子（TNF）-α诱导的血管内皮生长因子表达。在本研究中，我们检测了罗红霉素对TNF-α诱导的HPDL细胞产生MMP-1的影响。
将培养的细胞用1%胎牛血清孵育24小时，随后用10 ng/ml TNF-α、10 μM罗红霉素以及不同浓度的丝裂原活化蛋白激酶抑制剂进行处理。在不同时间点收集培养上清液和沉淀物，用于酶联免疫吸附测定、Northern印迹分析和Western印迹分析。
在HPDL细胞培养中，罗红霉素强烈抑制TNF-α诱导的MMP-1 mRNA表达和产生。罗红霉素对MMP-1基因表达的抑制依赖于从头合成蛋白质，且在转录水平受到调节。罗红霉素显著抑制TNF-α诱导的c-Jun氨基末端激酶激活（JNP），并轻微抑制细胞外信号调节激酶（ERK）1/2激活，但不抑制p38丝裂原活化蛋白激酶激活。此外，罗红霉素减少了MMP-1转录关键因子之一Ets-1的诱导。
罗红霉素通过下调ERK1/2和JNK激活以及随后减少Ets-1来抑制TNF-α介导的MMP-1诱导，提示罗红霉素可能在牙周炎和其他涉及MMP-1诱导的慢性炎症性疾病中具有治疗用途。
