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牙龈卟啉单胞菌脂多糖刺激下培养的人牙髓成纤维细胞白细胞介素-8基因表达

Interleukin-8 gene expression by human dental pulp fibroblast in cultures stimulated with Prevotella intermedia lipopolysaccharide.

作者信息

Nagaoka S, Tokuda M, Sakuta T, Taketoshi Y, Tamura M, Takada H, Kawagoe M

机构信息

Department of Operative Dentistry and Endodontology, Kagoshima University Dental School, Japan.

出版信息

J Endod. 1996 Jan;22(1):9-12. doi: 10.1016/S0099-2399(96)80228-7.

Abstract

Interleukin (IL)-8 mRNA expression was investigated in human dental pulp fibroblast cultures after stimulation with lipopolysaccharide (LPS) prepared from Prevotella intermedia and inflammatory cytokines. The expression of IL-8 mRNA and the release of IL-8 induced by P. intermedia LPS in pulpal fibroblast cultures were detected by Northern blot analysis and ELISA, respectively. The sufficient concentration of P. intermedia LPS on the IL-8 mRNA expression was 0.1 microgram/ml in pulpal fibroblast cultures. IL-8 mRNA levels began to increase after 2 h of exposure, reached a maximum at 4 to 8 h, and declined after 48 h, reaching the unstimulated level by 60 h. IL-8 production by the pulpal fibroblasts began to increase after 8 h of exposure upon stimulation with 10 microgram/ml of P. intermedia LPS. By contrast Salmonella LPS and synthetic lipid A did not increase IL-8 mRNA concentrations in pulpal fibroblast cultures. Recombinant human IL-1 alpha, beta, and tumor necrosis factor-alpha were capable of stimulating these cells to express IL-8 mRNA but natural human interferon-beta, gamma, and recombinant human IL-6 were incapable in our assay. These results suggest that pulpal fibroblasts are immunoresponsive cells and can elaborate IL-8 upon stimulation with P. intermedia LPS.

摘要

用中间普氏菌制备的脂多糖(LPS)和炎性细胞因子刺激人牙髓成纤维细胞培养物后,研究白细胞介素(IL)-8 mRNA的表达。分别通过Northern印迹分析和ELISA检测牙髓成纤维细胞培养物中IL-8 mRNA的表达以及中间普氏菌LPS诱导的IL-8释放。在牙髓成纤维细胞培养物中,中间普氏菌LPS对IL-8 mRNA表达的有效浓度为0.1微克/毫升。暴露2小时后,IL-8 mRNA水平开始升高,在4至8小时达到最大值,并在48小时后下降,到60小时达到未刺激水平。在用10微克/毫升中间普氏菌LPS刺激后,牙髓成纤维细胞暴露8小时后开始增加IL-8的产生。相比之下,沙门氏菌LPS和合成脂多糖不会增加牙髓成纤维细胞培养物中IL-8 mRNA的浓度。重组人IL-1α、β和肿瘤坏死因子-α能够刺激这些细胞表达IL-8 mRNA,但在我们的试验中,天然人干扰素-β、γ和重组人IL-6则无此作用。这些结果表明,牙髓成纤维细胞是免疫反应性细胞,在用中间普氏菌LPS刺激后能够产生IL-8。

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