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Effect of adalimumab on the expression profile of mRNA , and protein associated with JAK / STAT signaling pathway in fibroblast exposed to lipopolysaccharide
Author(s) -
Adwent Iwona,
Schweizer Michał,
Grabarek Beniamin O.,
Boroń Dariusz
Publication year - 2020
Publication title -
dermatologic therapy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.595
H-Index - 68
eISSN - 1529-8019
pISSN - 1396-0296
DOI - 10.1111/dth.13400
Subject(s) - adalimumab , lipopolysaccharide , gene silencing , tumor necrosis factor alpha , microbiology and biotechnology , messenger rna , receptor , inflammation , medicine , real time polymerase chain reaction , biology , immunology , gene , biochemistry
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.