Department of Oral Biology, Indiana University School of Dentistry, Indianapolis, IN 46202, USA.
J Periodontol. 2011 Oct;82(10):1504-8. doi: 10.1902/jop.2011.100757. Epub 2011 Feb 22.
Tobacco smoking is considered a major modifiable risk factor for periodontal disease. Nicotine is the addictive ingredient in tobacco and has been shown to affect multiple cellular processes. Neutrophils are the first line of host defense and are critical cells in the maintenance of periodontal health through their role in the control of bacteria, but they can also contribute to the progression of periodontal disease by the production and release of reactive oxygen species (ROS). Virulence factors from periodontal pathogens, such as Porphyromonas gingivalis (Pg), stimulate the respiratory burst of neutrophils. The objective of this study is to explore the oxidative activity of neutrophils when stimulated with Pg, nicotine, or both.
Neutrophils were separated from buffy coats by the double dextran gradient method. The generation of ROS by neutrophils was determined using luminol-dependent chemiluminescence assays. The reaction was followed for 90 minutes, and the neutrophil activation was recorded as the total integrated energy output.
The Pg and Pg plus nicotine groups had a significantly higher active and peak chemiluminescence than the nicotine group (all with P <0.0001). The Pg and Pg with nicotine groups were not significantly different (P = 0.90).
In the presence of Pg, the nicotine did not further enhance the ROS release by the neutrophils, suggesting that the bacteria induced the maximum ROS release in this model system.
吸烟被认为是牙周病的主要可改变危险因素。尼古丁是烟草中的成瘾成分,已被证明会影响多种细胞过程。中性粒细胞是宿主防御的第一道防线,通过控制细菌在维持牙周健康方面是关键细胞,但它们也可以通过产生和释放活性氧物质 (ROS) 来促进牙周病的进展。牙周病病原体(如牙龈卟啉单胞菌 (Pg))的毒力因子会刺激中性粒细胞的呼吸爆发。本研究旨在探讨 Pg、尼古丁或两者刺激中性粒细胞时的氧化活性。
通过双葡聚糖梯度法从白细胞层中分离中性粒细胞。使用发光依赖性化学发光测定法测定中性粒细胞产生的 ROS。反应持续 90 分钟,将中性粒细胞的激活记录为总积分能量输出。
Pg 组和 Pg 加尼古丁组的活性和峰值化学发光明显高于尼古丁组(均 P<0.0001)。Pg 组和 Pg 加尼古丁组之间无显著差异(P=0.90)。
在 Pg 存在的情况下,尼古丁并没有进一步增强中性粒细胞释放的 ROS,这表明在这个模型系统中,细菌诱导了最大的 ROS 释放。