Chen Yun-fang, Yan Jie, Zhang Di-ya, Chen Li-li
Department of Stomatology, The Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310009, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2008 Nov;37(6):622-8. doi: 10.3785/j.issn.1008-9292.2008.06.013.
To determine the effect of Porphyromonas gingivalis lipopolysaccharide (LPS) on induced secretion of inflammatory cytokines by different cell lines.
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 O111:B4 (E-LPS) was used as the control. TNF-alpha, IL-1beta, IL-6 and IL-8 levels in culture supernatants were measured by quantitative ELISA.
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-alpha and IL-1beta 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.
Pg-LPS can promote target cells to increase their secretion of inflammatory cytokines. KB cells can not be used as the target cell to determine inflammation-causing effect of LPS.
确定牙龈卟啉单胞菌脂多糖(LPS)对不同细胞系诱导分泌炎性细胞因子的影响。
采用酚水法提取牙龈卟啉单胞菌ATCC33277菌株的LPS(Pg-LPS),并通过鲎试剂法和红外光谱分析进行鉴定。用不同浓度和作用时间的Pg-LPS处理KB、HGF-1和THP-1细胞,以大肠杆菌O111:B4菌株的商品化LPS(E-LPS)作为对照。通过定量ELISA检测培养上清液中TNF-α、IL-1β、IL-6和IL-8的水平。
Pg-LPS和E-LPS使鲎试剂凝固的最小剂量均为15 ng/ml,且它们的红外光谱相似。用Pg-LPS或E-LPS处理后,HGF-1细胞分泌的TNF-α和IL-1β水平单次刺激时显著升高(P<0.01),而THP-1细胞的分泌则持续增强(P<0.01)。Pg-LPS或E-LPS均可刺激HGF-1或THP-1细胞持续增加IL-6的分泌(P<0.01)。Pg-LPS和E-LPS均可诱导THP-1细胞分泌IL-8(P<0.01),但只有Pg-LPS对HGF-1细胞有类似作用(P<0.01)。Pg-LPS和E-LPS均未诱导KB细胞分泌炎性细胞因子。
Pg-LPS可促进靶细胞增加炎性细胞因子的分泌。KB细胞不能用作确定LPS致炎作用的靶细胞。