Grabarek Beniamin, Wcisło-Dziadecka Dominika, Strzałka-Mrozik Barbara, Adamska Jolanta, Mazurek Urszula, Brzezińska-Wcisło Ligia
Department of Molecular Biology, School of Pharmacy with Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia ul. Jednosci 8, 41-200 Sosnowiec, Poland.
Department of Cosmetology, School of Pharmacy with Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, ul. Kasztanowa 3, 41-200 Sosnowiec, Poland.
Curr Pharm Biotechnol. 2018;19(15):1232-1240. doi: 10.2174/1389201020666190111163312.
The aim of the study was to evaluate the changes in the expression of genes - TNF-α, IFN-γ, depending on the time and concentration of IL-12 used to expose the Normal Human Dermal Fibroblast (NHDF) cells.
The material for the study included NHDF exposed to IL-12 in various IL-12 concentrations (1/10/100 ng/ml) and exposure time 0.5h, 1h, 2h, 4h, 8h, 24h, compared to the control. Changes in gene expression were evaluated with the use of the RTqPCR method. The statistical analysis was performed with the use of Statistica 12.5 PL The role of genes of the JAK-STAT signaling pathway in the induction of the inflammatory process was determined with the use of the PANTHER overrepresentation test.
Regardless of the time of NHDF exposure and the IL-12 concentrations used, we observed changes in the expressions of TNF-α and IFN-γ. Increased expression of one transcript involves the decreased expression of the other. We assessed that genes of the JAK-STAT signaling pathway are engaged in 6 biological processes, 8 molecular functions, 6 signaling pathways.
The conducted studies indicate that conclusions about the intensity of the inflammatory process, and the efficacy of the anti-cytokine therapeutic strategy, may be made on the basis of the TNF-α, IFN-γ expression.
本研究的目的是评估基因 - TNF-α、IFN-γ 的表达变化,这取决于用于处理正常人皮肤成纤维细胞(NHDF)的白细胞介素 - 12(IL-12)的时间和浓度。
研究材料包括以不同浓度(1/10/100 ng/ml)的 IL-12 处理 0.5 小时、1 小时、2 小时、4 小时、8 小时、24 小时的 NHDF,与对照组进行比较。使用 RTqPCR 方法评估基因表达的变化。使用 Statistica 12.5 PL 进行统计分析。使用 PANTHER 过表达测试确定 JAK-STAT 信号通路基因在炎症过程诱导中的作用。
无论 NHDF 的暴露时间和所使用的 IL-12 浓度如何,我们都观察到 TNF-α 和 IFN-γ 的表达变化。一种转录本表达的增加涉及另一种转录本表达的降低。我们评估 JAK-STAT信号通路的基因参与了 6 个生物学过程、8 种分子功能、6 条信号通路。
进行的研究表明,可以基于 TNF-α、IFN-γ 的表达得出关于炎症过程强度和抗细胞因子治疗策略疗效的结论。