Stadnyk A W, Sisson G R, Waterhouse C C
Department of Pediatrics, Dalhousie University, Halifax, Nova Scotia, Canada.
Exp Cell Res. 1995 Oct;220(2):298-303. doi: 10.1006/excr.1995.1319.
The rat small intestinal epithelial cell line, IEC-6, was examined for the presence of IL-1 mRNAs using the reverse transcription/polymerase chain reaction method. IL-1 alpha but not IL-1 beta transcripts were detected in plastic adherent cells. The levels of IL-1 alpha transcripts were similar in cells cultured at different densities. IL-1 activity was not detected, by bioassay, in the culture supernatant of the cells, nor was it membrane associated. IL-1 activity was detected in cell lysates, although its measurement was made difficult by the presence of an inhibitor of the bioassay. Intracellular IL-1 alpha was detectable using Western blots. Interleukin-1 receptor antagonist mRNA was also detectable in plastic adherent cells. Treatment with TGF-beta 1, IL-1 beta, IL-6, TNF-alpha, or combinations of any two of these cytokines failed to induce the secretion of the IL-1 alpha or to significantly change the levels of mRNA. The IEC-6 cell line is similar to other epithelial cells in having intracellular pools of IL-1 alpha.
利用逆转录/聚合酶链反应方法检测大鼠小肠上皮细胞系IEC-6中白细胞介素-1（IL-1）mRNA的存在情况。在贴壁于塑料培养皿的细胞中检测到了IL-1α转录本，但未检测到IL-1β转录本。不同密度培养的细胞中IL-1α转录本水平相似。通过生物测定法，在细胞培养上清液中未检测到IL-1活性，其也不与细胞膜相关。在细胞裂解物中检测到了IL-1活性，尽管生物测定法中存在抑制剂使得其测量变得困难。使用蛋白质免疫印迹法可检测到细胞内的IL-1α。在贴壁于塑料培养皿的细胞中也可检测到白细胞介素-1受体拮抗剂mRNA。用转化生长因子-β1（TGF-β1）、IL-1β、IL-6、肿瘤坏死因子-α（TNF-α）或这些细胞因子中任意两种的组合进行处理，均未能诱导IL-1α的分泌，也未显著改变mRNA水平。IEC-6细胞系与其他上皮细胞类似，具有细胞内IL-1α库。
