Wendell K J, Stein S H
University of Tennessee Health Science Center, College of Dentistry, Department of Periodontology, Memphis 38163, USA.
J Periodontol. 2001 Aug;72(8):1038-44. doi: 10.1902/jop.2001.72.8.1038.
Patients who smoke are at increased risk for chronic periodontitis (CP). Most studies suggest that the microbial flora in these patients is similar to that found in non-smoking CP patients. Thus, the increased risk for development of CP is not dependent on an altered microbial profile, but rather to some change in the host response to these periopathogens. There is evidence that human gingival fibroblasts (HGF) derived from diseased sites produce greater amounts of interleukin (IL)-6 and IL-8 in vitro than cells derived from healthy sites. This suggests that HGF subpopulations may be selected based upon the inflammatory milieu in which they reside. The hypothesis to be tested was that the combination of nicotine and lipopolysaccharide (LPS) could regulate HGF inflammatory mediator production.
HGF cell cultures were established from explants derived from 10 patients with CP. HGF cell cultures were stimulated with 1 mM, 1 microM, or 1 nM nicotine +/- Escherichia coli or Porphyromonas gingivalis LPS. At 12, 24, or 48-hour time points, the cells were counted and the supernatant was collected for subsequent IL-6 and IL-8 determination in an enzyme-linked immunosorbent assay.
At the 24-hour time point, 1 nM nicotine stimulated IL-6 production compared to control (P=0.02). E. coli LPS alone caused a 3- to 4-fold increase in IL-6 and IL-8 production, whereas P gingivalis LPS did not augment IL-6 or IL-8. A synergistic effect upregulating IL-6 was observed with combined treatment of 1 mM nicotine and E. coli LPS or P gingivalis LPS at the 24-hour time point (P<0.0005 and P=0.002, respectively). Similar effects were seen when IL-8 production was evaluated following HGF stimulation with high doses of nicotine and E. coli LPS or P gingivalis LPS.
These results demonstrate that nicotine by itself can stimulate HGF IL-6 and IL-8 production. Moreover, the combination of high doses of nicotine and either E. coli or P gingivalis LPS can synergistically upregulate cytokine production. These findings support the hypothesis that a proinflammatory fibroblast phenotype may be elicited in an environment enriched with bacterial LPS and nicotine.
吸烟患者患慢性牙周炎（CP）的风险增加。大多数研究表明，这些患者的微生物菌群与非吸烟CP患者的相似。因此，CP发病风险增加并非取决于微生物谱的改变，而是宿主对这些牙周病原体反应的某些变化。有证据表明，来自患病部位的人牙龈成纤维细胞（HGF）在体外比来自健康部位的细胞产生更多的白细胞介素（IL）-6和IL-8。这表明HGF亚群可能根据它们所处的炎症环境进行选择。有待检验的假设是尼古丁和脂多糖（LPS）的组合可调节HGF炎症介质的产生。
从10例CP患者的外植体建立HGF细胞培养物。用1 mM、1 microM或1 nM尼古丁±大肠杆菌或牙龈卟啉单胞菌LPS刺激HGF细胞培养物。在12、24或48小时时间点，对细胞进行计数，并收集上清液用于随后的酶联免疫吸附测定中IL-6和IL-8的测定。
在24小时时间点，与对照相比，1 nM尼古丁刺激IL-6产生（P = 0.02）。单独的大肠杆菌LPS使IL-6和IL-8产生增加3至4倍，而牙龈卟啉单胞菌LPS并未增加IL-6或IL-8。在24小时时间点，1 mM尼古丁与大肠杆菌LPS或牙龈卟啉单胞菌LPS联合处理观察到上调IL-6的协同效应（分别为P < 0.0005和P = 0.002）。在用高剂量尼古丁和大肠杆菌LPS或牙龈卟啉单胞菌LPS刺激HGF后评估IL-8产生时也观察到类似效果。
这些结果表明，尼古丁本身可刺激HGF产生IL-6和IL-8。此外，高剂量尼古丁与大肠杆菌或牙龈卟啉单胞菌LPS的组合可协同上调细胞因子的产生。这些发现支持了在富含细菌LPS和尼古丁的环境中可能引发促炎成纤维细胞表型的假设。
