IL-1α和肿瘤坏死因子α刺激牙龈成纤维细胞产生IL-11的研究

Production of interleukin-11 in human gingival fibroblasts stimulated with IL-1α and tumor necrosis factor-α

目的 为了解白细胞介素 (IL) 11在牙周疾病中的作用 ,检测IL 1α和肿瘤坏死因子α(TNF α)刺激牙龈成纤维细胞 (HGFs)IL 11的表达和调控。方法 收集细胞上清液 ,ELISA法测定IL 11的水平 ;提取细胞mRNA ,实时定量RT PCR法定量分析IL 11mRNA和看守基因GAPDHmRNA的表达。结果 IL 1α和TNF α均可显著增加HGFs表达的IL 11水平 (P <0 0 1) ,两者还有明显的协同刺激效应 ,产生的IL 11远高于其单独刺激HGFs产生的IL 11之和 ;吲哚美辛显著抑制了IL 1α和TNF α单独或联合刺激的HGFsIL 11的产生 (P <0 0 1)。HGFs受到IL 1α刺激 6h即有明显的IL 11mRNA(IL 11/GAPDH比值 3 6 )表达 ,同时加入放线菌酮后 ,IL 11mRNA的表达则明显减弱 (IL 11/GAPDH比值 2 4 ) ;2 4h时 ,HGFs受到IL 1α刺激IL 11mRNA表达继续增强 (IL 11/GAPDH比值 5 4 ) ,与TNF α联合刺激IL 11mRNA表达增强最显著 (IL 11/GAPDH比值 8 7) ,但都受到吲哚美辛的显著抑制 (IL 11/GAPDH比值小于 1)。结论 IL 1α和TNF α刺激HGFs显著增加IL 11的产生 。

Objective To investigate what role IL-11 plays in periodontal disease and to determine the level of IL-11 in HGFs stimulated with IL-1α and TNF-α Methods HGFs were stimulated with IL-1α and TNF-α alone or in combination The production of IL-11 was measured using enzyme-linked immunosorbent assay (ELISA) IL-11 and Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) messenger RNA (mRNA) levels in HGFs were determined by real-time reverse transcription-polymerase chain reaction(RT-PCR) Results IL-1α significantly increased the levels of IL-11 in HGFs TNF-α also significantly augmented IL-11 production in HGFs, and synergistically stimulated HGFs to produce IL-11 when combined with IL-1α Indomethacin, an inhibitor of prostaglandin synthesis, significantly reduced IL-11 production by HGFs stimulated with IL-1α and TNF-α individually or in combination IL-1α alone or combined with TNF-α enhanced the ratio of IL-11/GAPDH mRNA expression in HGFs, and the augmentation was abolished by indomethacin after co-incubation for 24 hs Conclusions Production of IL-11 in HGFs stimulated with IL-1α and TNF-α was transcriptionally upregulated by the endogenous prostaglandin synthesis Inhibition of prostaglandin might suppress the osteoclastogenesis by IL-11 in inflammatory periodontal diseases