内毒素耐受对人正常前列腺上皮细胞分泌IL-6和IL-8的影响

The influence of endotoxin tolerance on the expression of IL-6 and IL-8 in normal human prostate epithelial cells

目的 研究发现前列腺组织炎症在前列腺增生的发生与进展中有着重要作用.方法 内毒素耐受（endotoxin tolerance,ET）现象存在于人组织上皮细胞中如肠上皮细胞和人牙龈上皮细胞等,并可能影响组织的炎症和免疫反应.产生内毒素/脂多糖（Lipopolysaccharide,LPS）的G-菌如大肠埃希菌是影响前列腺炎的主要病原体.因此,本研究拟通过分析LPS反复刺激人正常前列腺上皮细胞RWPE-1中细胞炎性因子IL-6和IL-8表达,探讨是否内毒素耐受现象存在于人前列腺上皮细胞中.结果 采用1μg/mL大肠杆菌（Escherichia coli） LPS刺激人正常前列腺上皮细胞RWPE-1 24 h,洗涤细胞后,给予1μg/mL浓度LPS再次刺激24 h,构建内毒素耐受模型.采用液相芯片技术检测细胞培养液中细胞因子IL-6和IL-8分泌水平的变化.结果LPS刺激后6h和24 h,IL-6和IL-8分泌水平均呈现上升趋势（P＜0.05）,较空白对照组有明显增高（P＜0.05）,LPS反复刺激后,诱导细胞耐受后,IL-6和IL-8分泌水平较第1次刺激后明显降低（P＜0.05）.结论 内毒素耐受现象存在于RWPE-1中,能抑制RWPE-1细胞因子IL-6和IL-8的表达,进而可能影响前列腺组织的炎症和免疫反应.

Objective Current studies have shown that tissue inflammation plays an important role in the course of development of benign prostate hyperplasia （BPH）.Endotoxin tolerance （ET） was found to exist in human tissue cells,such as intestinal epithelial cells and human gingival epithelial cells,and could affect the tissue inflammation and immune responses.Lipopolysaccharide （LPS） generated by G-bacteria such as Escherichia coli,which was considered as the most important pathogen of tissue inflammation of prostate.We intended to analyze the inflammatory cytokines interleukin -6 （IL-6） and interleukin-8 （IL-8） expression of normal human prostate epithelial cells RWPE -1 which were stimulated by LPS repeatedly to find out whether ET exist in human prostate epithelial cells as well.Methods RWPE-1 cells were given stimulation of 1 μg/mL LPS which came from Escherichia coli for 24 hours.After collected the culture supernatant of REWP-1,1 μg/mL LPS group stimulation was given again to form a model of ET.Expression level of cytokines IL-6 and IL-8 was detected by Millipore human cytokine detection kit instructions,in Luminex platform.Results Expression of IL-6 and IL-8 was significant rising at 6 hours and 12 hours after first LPS stimulation （P ＜ 0.05）.Compared with first LPS stimulation,expression of IL-6 and IL-8 was significant lower after stimulated by LPS repeatedly （P ＜ 0.05）.Conclusions Endotoxin tolerance phenomenon exists in normal prostate epithelial cells RWPE-1,and inhibits the expression of IL-6 and IL-8.It may influence the chronic inflammation and immune response of prostate tissue.