Adwent Iwona, Schweizer Michał, Grabarek Beniamin O, Boroń Dariusz
Department of Histology, Cytophysiology and Embryology, Faculty of Medicine in Zabrze, University of Technology in Katowice, Katowice, Poland.
Department of Dermatology, Andrzej Mielecki Memorial Independent Public Clinical Hospital, Medical University of Silesia, Katowice, Poland.
Dermatol Ther. 2020 May;33(3):e13400. doi: 10.1111/dth.13400. Epub 2020 Apr 29.
The aim of this study was to assess the effect of adalimumab on the expression level of mRNA and protein TNF-α, IFN-γ, IL-17, IL12A, IL12B, and IL23A in the culture of normal human fibroblasts, in which the LPS inflammation process was induced. The NHDF culture was exposed to the effect of LPS in the concentrations of 1, 2, and 10 μg/mL for 2, 8, and 24 hour periods, and then adalimumab was added at the concentration of 8 μg/mL, it was then incubated for 2, 8, and 24 hour. Cells unexposed to LPS and adalimumab constituted the control. The microarray expression techniques, RTqPCR, and ELISA assay were used. Irrespectively of the concentration of LPS used and the incubation time of it with cells overexpression of the analyzed genes is present, with increasing factor concentration used to induce inflammation and incubation time with it, expression of the assessed genes was greater. In turn, adding the anti-TNF drug to the culture caused the silencing of the expression of the mRNAs and proteins. It was confirmed that LPS and adalimumab above all affect the expression of genes and proteins dependent on the interaction of IL-12 with receptors, which are TNF-α and IFN-γ, and to a lesser extent also modulate IL-17.
本研究的目的是评估阿达木单抗对诱导脂多糖（LPS）炎症过程的正常人成纤维细胞培养物中mRNA和蛋白肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）、白细胞介素-17（IL-17）、白细胞介素12A（IL12A）、白细胞介素12B（IL12B）和白细胞介素23A（IL23A）表达水平的影响。将正常人皮肤成纤维细胞（NHDF）培养物分别暴露于浓度为1、2和10μg/mL的LPS中2、8和24小时，然后加入浓度为8μg/mL的阿达木单抗，再孵育2、8和24小时。未暴露于LPS和阿达木单抗的细胞作为对照。使用了微阵列表达技术、逆转录定量聚合酶链反应（RTqPCR）和酶联免疫吸附测定（ELISA）。无论所用LPS的浓度以及其与细胞的孵育时间如何，所分析基因均存在过表达，随着用于诱导炎症的因子浓度增加及其孵育时间延长，所评估基因的表达更高。反过来，向培养物中添加抗TNF药物导致mRNA和蛋白表达沉默。已证实，LPS和阿达木单抗首先影响依赖于IL-12与受体（即TNF-α和IFN-γ）相互作用的基因和蛋白的表达，并且在较小程度上也调节IL-17。
