Pro-inflammatory cytokine production from normal human fibroblasts is induced by Tannerella forsythia detaching factor.
Pro-inflammatory cytokine production from normal human fibroblasts is induced by Tannerella forsythia detaching factor.
复制标题
正常人成纤维细胞产生促炎细胞因子是由连翘分离因子诱导的。
DOI：
10.1111/j.1600-0765.2007.01003.x
复制
发表时间：
2008
影响因子：
3.5
通讯作者：
T. Nakajima
中科院分区：
文献类型：
--
作者：
N. Tomi;Y. Fukuyo;Shinichi Arakawa;T. Nakajima
背景和目标
连翘坦纳氏菌是一种牙周病原菌。最近，我们报道了连翘的细胞病变成分包含两个不同的因子。一种是将细胞周期停滞在G2期，另一种称为连翘分离因子，它可以分离黏附依赖的永生化人类细胞。在这项研究中，我们利用人正常成纤维细胞来研究连翘分离因子的生物学功能。
材料和方法
使用了先前报道的重组连翘分离因子。TIG-3细胞在无或有连翘分离因子的条件下培养，裂解培养上清液，用多克隆连翘分离因子抗体进行免疫印迹分析。随后对细胞进行分离，以分离细胞质、线粒体和剩余的部分。为了用烟酰胺腺嘌呤二核苷酸连接还原酶检测线粒体的活性，采用了水溶四氮唑法。用流式细胞术测定二氢四甲基罗胺氧化依赖的荧光转化率来估算线粒体膜氧化电位。用人IL-8酶联免疫吸附试剂盒检测培养上清液中IL-8的浓度。
结果
经连翘脱附因子处理的细胞在3~24 h内脱离基质并聚集，48h后细胞恢复黏附并增殖，Western印迹分析显示大部分连翘脱附因子转位到线粒体部分。连翘分离因子以剂量依赖的方式抑制烟酰胺腺嘌呤二核苷酸连接还原酶的活性，从而增加线粒体膜氧化电位。在72小时，连翘分离因子处理的细胞通过增加线粒体氧化膜电位来增强白介素8的产生。
结论
连翘分离因子可能通过诱导促炎细胞因子IL-8参与连翘毒力的形成。
BACKGROUND AND OBJECTIVE
Tannerella forsythia is a periodontal pathogen. Recently, we have reported that the cytopathic component of T. forsythia contains two distinct factors. One arrests the cell cycle at the G2 phase and the other, named forsythia detaching factor, detaches adhesion-dependent immortalized human cells. In this study, we investigated the biological function of forsythia detaching factor using human normal fibroblasts.
MATERIAL AND METHODS
A recombinant forsythia detaching factor, reported previously, was used. TIG-3 cells, cultured in the absence or presence of forsythia detaching factor, were lysed and the supernatant was analyzed by western blotting with polyclonal forsythia detaching factor antibodies. The cells were subsequently fractionated to isolate the cytoplasmic, mitochondrial and remaining fractions. In order to measure the activity of mitochondria using nicotinamide adenine dinucleotide-linked reductase, the water-soluble tetrazolium method was used. The mitochondrial oxidative membrane potential was estimated by measuring the oxidization-dependent fluorogenic conversion of dihydrotetramethylrosamine using flow cytometry. The concentration of interleukin-8 in the culture supernatant was assayed using a Human IL-8 ELISA kit.
RESULTS
Forsythia detaching factor-treated cells detached from the substratum and aggregated from 3 to 24 h. Then, the detached cells resumed adhesion and proliferated after 48 h. The western blot analysis revealed that most forsythia detaching factor trans-located into the mitochondrial fraction. Forsythia detaching factor suppressed the nicotinamide adenine dinucleotide-linked reductase activity in a dose-dependent manner and consequently increased the mitochondrial oxidative membrane potential. The production of interleukin-8 was reinforced in forsythia detaching factor-treated cells at 72 h through an increase of the mitochondrial oxidative membrane potential.
CONCLUSION
The forsythia detaching factor might be involved in the virulence of T. forsythia through induction of the pro-inflammatory cytokine interleukin-8.
影响因子：
6.7
作者：
Socransky, SS;Haffajee, AD;Kent, RL
通讯作者：
Kent, RL