The Influence of Adalimumab and Cyclosporine A on the Expression Profile of the Genes Related to TGFβ Signaling Pathways in Keratinocyte Cells Treated with Lipopolysaccharide A

Abstract

Background. In the treatment of moderate to severe psoriasis, cyclosporine A (CsA) conventional therapy is used and biological, anti-cytokine treatment using, for example, anti-TNF drug—adalimumab. Aim. This study aimed at investigating the effect of CsA and adalimumab on the profile of mRNAs and protein expression associated with transforming growth factor β (TGFβ) pathways in human keratinocyte (HaCaT) culture previously exposed to lipopolysaccharide (LPS). Materials and Methods. HaCaT culture was exposed to 1 ng/ml LPS for 8 hours+8 μg/ml adalimumab for 2, 8, and 24 hours or 1 ng/ml LPS for 8 hours+100 ng/ml CsA for 2, 8, and 24 hours and compared to the control culture. Sulphorodamine B cytotoxicity assay was performed. The expression profile of mRNA related to TGFβ paths was indicated by microarray and RTqPCR analyses. The ELISA test was used to analyze changes on the proteome level. Statistical analysis consisted of ANOVA analysis and the post hoc Tukey test (p<0.05). Results. The cytotoxicity test showed that LPS, adalimumab, and cyclosporine in the concentration used in this experiment did not have any cytotoxicity effect on HaCaT cells. The largest fold changes (FC) in expression in (∣FC∣>4.00) was determined for TGFβ1-3, TGFβRI-III, SKIL, SMURF2, SMAD3, BMP2, BMP6, JAK2, UBE2D1, SKP2, EDN1, and PRKAR2B (p<0.05). In addition, on the protein level, the direct changes observed at mRNA were the same. Conclusion. Analysis of the microarray expression profile of genes associated with TGFβ signaling pathways has demonstrated the potential of cyclosporin A and adalimumab to induce changes in their transcriptional activity. The anti-TNF drug seems to affect TGFβ cascades to a greater extent than cyclosporin A. The obtained results suggest that the regularity of taking the drug is important for the efficacy of psoriasis therapy.

Evaluation of the Influence of Adalimumab on the Expression Profile of Leptin-Related Genes and Proteins in Keratinocytes Treated with Lipopolysaccharide A

“…In their work, Emilio-Silvia et al show that a mixture of terpenes in the form of citral lowers the concentration of leptin and IL-6, TNF-α in the serum of mice, where a systemic inflammatory response was induced by LPS [ 41 ]. These results confirm the fact that the JAK/STAT signaling pathway, whose final products are TNF-α and IL-6, is a key signaling cascade activated by the interactions between leptin and its receptors [ 19 , 46 ].…”

“…Taking into account, our previous studies, keratinocytes were incubated with 1 μg/mL LPS for 8 h [ 20 ] and next with 8 μg/mL adalimumab. The concentration of this drug corresponds to the concentration in the serum of psoriatic patients [ 19 ].…”

“…This was a reason for using TNF-α inhibitors in therapy of moderate and acute psoriasis. One of these is adalimumab which exhibits the ability to bind and inactivate membrane and soluble form of TNF-α [18], which in turn, as we have observed in previous studies, has an effect on the JAK/STAT cascade [19]. In in vitro studies in order to replicate inflammatory conditions in keratinocytes altered by psoriasis, lipopolysaccharides A are used [20].…”

Genetic Disruption of Guanylyl Cyclase/Natriuretic Peptide Receptor-A Triggers Differential Cardiac Fibrosis and Disorders in Male and Female Mutant Mice: Role of TGF-β1/SMAD Signaling Pathway

“…The activation of the TGF-β1 signaling pathway has diverse physiological roles, including morphogenesis, cell growth, development, and survival [ 27 , 31 , 32 , 33 , 46 ]. In the heart, TGF-β1 activates fibroblast proliferation, resulting in progressive fibrosis [ 44 , 45 , 47 ].…”

“…A number of fibrogenic factors seem to regulate cardiomyocyte growth, fibroblast activation, and extracellular matrix (ECM) deposition. TGF-β1 is a major profibrotic agent, which drives cardiac cellular signaling in hypertrophic and fibrotic conditions and is known to exert pleiotropic effects in the regulation of critical cellular and biological processes [ 27 , 28 , 29 ]. The members of the TGF-β1 superfamily influence the diverse nature of cellular processes, including cell proliferation and differentiation as well as programmed cell apoptosis and death [ 30 , 31 , 32 , 33 , 34 ].…”