体外培养人肠上皮细胞株接受脂多糖联合热打击后白细胞介素6、肿瘤坏死因子a的释放  

Effect of lipopolysaccharide stimulation combined with heat stress on release of interleukin-6 and tumor necrosis factor-alpha from human intestinal epithelial cells cultured in vitro

作　　者：孙杰[1] 陈蕊[1] 董家辉[1] 姜巧[1] 孙照琨[1] 郭振辉[1] 

机构地区：[1]解放军广州军区广州总医院MICU科,广东省暨广州市老年感染与器官功能支持重点实验室,广东省广州市510010

出　　处：《中国组织工程研究》2014年第51期8238-8242,共5页Chinese Journal of Tissue Engineering Research

基　　金：广州市科技计划项目(2014J4100033);解放军总后勤部专项基金(132000273);课题名称:老年脓毒症功能障碍的临床评估与干预~~

摘　　要：背景:对于肠上皮细胞遭受高热和肠道细菌以及由细菌产生的内毒素等多重打击后经历了怎样的分子生物学变化,一直缺乏切实、可靠实验依据。目的:观察脂多糖联合热打击对人肠上皮细胞损伤及炎症因子白细胞介素6、肿瘤坏死因子肿瘤坏死因子a释放的影响。方法:体外培养人肠上皮细胞株(SW 480),将3×105细胞种入实验用培养皿。实验分组为:对照组(37℃),热打击组(43℃2 h),脂多糖刺激组(1 mg/L脂多糖6 h),热打击联合脂多糖刺激组(43℃+脂多糖)。采用WST-1法观察细胞毒效应和细胞存活率;同时采用ELISA法观察各组炎症因子白细胞介素6和肿瘤坏死因子a水平。结果与结论:与对照组相比,热打击组具有细胞毒效应,其细胞活率明显下降,细胞炎症因子白细胞介素6和肿瘤坏死因子a水平也有所增高。脂多糖组单独作用也具有细胞毒效应,细胞活力、细胞增殖率也明显下降,同时白细胞介素6和肿瘤坏死因子a水平明显升高。而热打击和脂多糖联合打击后,可观察到两者的协同作用。结果说明,热打击和脂多糖单独作用均可引起细胞损伤,增加炎症因子白细胞介素6和肿瘤坏死因子a表达,并且二者之间存在协同效应。实验结果为理解重症中暑病理过程中,肠道功能障碍、肠道细菌和内毒素移位进而引发多脏器功能衰竭的病理机制提供了客观的实验依据。BACKGROUND： There is no practical and reliable experimental evidence for changes in molecular biology of intestinal epithelial cells suffering from heat stress, intestinal bacteria and endotoxin produced by bacteria, OBJECTIVE： To observe the effect of lipopolysaccharide stimulation combined with heat stress on human intestinal epithelial cell injury and release of interleukin-6 and tumor necrosis factor-alpha. METHODS： Human intestinal epithelial cell lines （SW 480） cultured in vitro was seeded into culture dishes at a density of 3×105, and then divided randomly into four groups： control group was cultured in an incubator of 37 ℃ heat stress group was cultured in the incubator of 43 ℃ for 2 hours; lipopolysaccharide group was stimulated by 1 mg/L lipopolysaccharide for 6 hours; lipopolysaccharide＋heat stress group （combination group） was subjected to heat stress for 2 hours followed by lipopolysaccharide stimulation for 6 hours. WST-1 method was used to investigate the cytotoxic effect and cell survival rate, and ELISA assay was used to investigate the levels of interleukin-6 and tumor necrosis factor-alpha. RESULTS AND CONCLUSION： Compared with the control group, the cytotoxic effect increased, and the cell survival rate reduced as well as the level of interleukin-6 and tumor necrosis factor-alpha increased in the heat stress and lipopolysaccharide groups. Synergistic effects were visible in the combination group. These findings indicate that lipopolysaccharide and heat stress alone can both induce cell injury, increase the levels of interleukin-6 and tumor necrosis factor-alpha, and there is a synergistic effect between them. Experimental findings from this study may be an objective experimental basis for understanding the pathogenesis of multiple organ failure caused by bowel dysfunction, intestinal bacteria and endotoxin translocation during the pathological process of severe heat stroke.

关 键 词：脂多糖类 上皮细胞 白细胞介素6 肿瘤坏死因子A 组织构建 组织工程 脂多糖 热打击 肠上皮细胞 炎症因子 

分 类 号：R318[医药卫生—生物医学工程]