摘要
目的:探讨牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)脂多糖(LPS),诱导人口腔上皮细胞KB和人牙龈成纤维细胞HGF-1及人单核-巨噬样细胞THP-1,分泌炎性细胞因子的能力及其差异。方法:采用酚水法提取PgATCC33277株的脂多糖(Pg-LPS)。采用鲎试验和红外光谱对提取的Pg-LPS进行鉴定。采用ELISA试剂盒,定量检测不同浓度和时间Pg-LPS作用的上述3种细胞培养物上清中TNF-α、IL-1β、IL-6和IL-8变化水平。实验中采用商品化大肠杆菌O111:B4脂多糖(E-LPS)为对照。结果:Pg-LPS和E-LPS凝固鲎试剂所需最低浓度均为15ng/ml,其红外光谱也极为相似。在Pg-LPS或E-LPS作用下,HGF-1细胞分泌的TNF-α和IL-1β水平呈单峰形增高(P<0.01),但THP-1细胞为持续性升高(P<0.01)。Pg-LPS或E-LPS具有促HGF-1和THP-1细胞持续性增强IL-6分泌的作用(P<0.01)。Pg-LPS和E-LPS均可诱导THP-1细胞增强IL-8的分泌(P<0.01),但对HGF-1细胞仅Pg-LPS有促IL-8分泌的效应(P<0.01)。Pg-LPS和E-LPS均不能诱导KB细胞分泌上述细胞因子。结论:Pg-LPS有很强的促靶细胞分泌多种炎性细胞因子的生物学活性,从而引发初始的局部炎症反应。Pg-LPS致炎作用与E-LPS相似,但诱导不同炎性细胞因子的模式颇有差异。KB细胞不能作为LPS致炎作用的效应靶细胞。
Objective： To determine the effect of Porphyromonas gingivalis lipopolysaccharide （LPS） on induced secretion of inflammatory cytokines by different cell lines. Methods： LPS of P. gingivalis strain ATCC33277 （Pg-LPS） was extracted with phenol-water method, and identified by Limulus test and infrared spectrum analysis. KB, HGF-1 and THP-1 cells were treated with Pg-LPS of different concentrations and time duration, a commercial LPS of E. coli strain Olll .B4 （E-LPS） was used as the control. TNF-α, IL-1β, IL-6 and IL-8 levels in culture supernatants were measured by quantitative ELISA. Results. The minimal dosages of both Pg -LPS and E-LPS to solidify Limulus agents were 15 ng/ml and their infrared spectrums were similar. With the treatment of Pg-LPS or E-LPS, the TNF-α and IL-1β levels secreted by HGF -1 cells were remarkably increased with a single perk （P〈0.01） while a continuous enhancement of secretion by THP-1 cells was observed （P〈0.01）. Either Pg-LPS or E-LPS stimulated HGF-1 or THP-1 cells to continuously increase the secretion of IL-6 （P〈0. 01）. Both Pg-LPS and E-LPS induced IL-8 secretion by THP-1 cells （P〈0. 01）, but only Pg-LPS showed the similar effect on HGF-1 cells （P〈0. 01）. Neither Pg-LPS nor E-LPS induced KB cells to secrete inflammatory cytokines. Conclusion. Pg-LPS can promote target cells to increase their secretion of inflammatory cytokines. KB ceils can not be used as the target cell to determine inflammation -causing effect of LPS.
出处
《浙江大学学报（医学版）》
CAS
CSCD
2008年第6期622-628,633,共8页
Journal of Zhejiang University(Medical Sciences)
基金
国家自然科学基金资助项目(30471888)
关键词
脂多糖类/药理学
牙龈卟啉单胞菌
致炎作用
细胞因子
Lipopolysaccharides/pharmacol
Porphyromonas gingivalis
Inflammation causing effect
Cytokines
