脂多糖对原代大鼠腹膜间皮细胞IL-1β和IL-6表达的影响

Effect of lipopolysaccharide on the expressions of IL-1β and IL-6 in rat peritoneal mesothelial cells

目的研究原代腹膜间皮细胞(PMCs)对不同浓度脂多糖(LPS)刺激的反应效果,探讨术后腹腔粘连体外细胞最佳造模方法。方法运用不同浓度LPS于不同时间点刺激PMCs,ELISA法检测各组细胞上清液中白介素(IL)-1β、IL-6蛋白含量,qRT-PCR检测IL-1β、IL-6 mRNA水平,扫描电镜观察LPS刺激前后PMCs的活性变化情况。结果 ELISA测得各组大鼠PMCs培养上清液IL-1β和IL-6浓度随着培养时间推移和LPS浓度增加而表现出明显差异(P<0.01),尤其以24 h、10μg/ml为LPS最佳刺激时间和最佳刺激浓度;qRT-PCR实验中LPS最佳刺激时间为24 h,但是最佳刺激浓度为5μg/ml;正常PMCs扫描电镜下表面被丰富的微绒毛覆盖;LPS刺激后PMCs剥脱、肿胀明显,具有细胞间隙,偶见基底膜。结论LPS能够刺激大鼠PMCs诱发炎症损伤,进一步放大炎症反应,模拟术后腹腔粘连微环境,作为术后腹腔粘连体外细胞模型。

Objective To investigate the stimulating effects of lipopolysacchride（ LPS） on the expressions of IL-1β and IL-6 in rat peritoneal mesothelial cells（ RPMCs）. Methods Different concentrations of LPS was used to stimulate RPMCs at different time points,the protein and mRNA levels of IL-Iβ,IL-6 were detected respectively by ELISA and qRT-PCR. Morphological changes were observed under scanning electron microscope. Results IL-Iβ and IL-6 concentrations were increased with the stimulation time and LPS concentration（ P〈0.01）. The best stimulation time was 24 hours and the best stimulus concentration was 10 μg / ml. Normal RPMCs manifestations were covered by abundant microvilli. However,LPS stimulated RPMCs appear stripping,swelling significantly,with the gap and the basement membrane.Conclusions RPMCs could induce inflammation injury stimulated by LPS,further amplifying inflammatory response.