The article prepared for publication in the “Journal of Biomedical Research and Therapeutics” describes for the first time the effect of TFX on inhibition of viral replication, suppression of pro-inflammatory cytokine production and expression of mitogen-activated kinases in lymphoid organs. In addition, some of the previously known properties of the drug were confirmed.
Several in vitro studies are described in this article. The first was to check the cytotoxicity of TFX, which turned out to be non-harmful to cells, including bone marrow cells. The effect of TFX on concanavalin A (ConA) induced thymocyte proliferation was then examined. As a result of the study, it was observed that TFX has a positive effect on cell proliferation, especially in the conditions of low cell density and optimal ConA concentration. In addition, it was investigated how ConA and TFX can restore the ability to proliferate lost as a result of, for example, mental stress or the influence of steroids. Thymocytes exposed to hydrocortisone proliferated significantly worse, and after overnight incubation in the substances mentioned above, this ability returned, although not to the maximum level.
The next part of the study was the observation of the effect of TFX on the production of cytokines by macrophages under the influence of lipopolysaccharide (LPS). LPS is a bacterial toxin found in the outer cell membrane of Gram-negative bacteria and cyanobacteria. It was checked how macrophages will react when they are first exposed to TFX and then LPS and when both substances are administered simultaneously. In the first part, there was no inhibition in the production of IL-1b, and a 10% decrease in the production of TNFa and an almost 20% decrease in the production of IL-6. Co-administration of LPS and TFX increased the inhibitory effect of the drug and macrophages produced 20% less TNFa, 10% less IL-6 and approximately 25% less IL-1b.
The antiviral effect of TFX was checked in two ways: direct contact of the preparation with the herpes simplex virus and its administration to infected cells. In both cases, the higher drug concentration had a better effect and at 50 µg/ml it reduced viral replication by 100-fold.
The final part was to investigate the effects of TFX on Mitogen Activated Protein Kinases (MAPKs) in both the lymphoid organs – bone marrow, thymus and spleen, and in four cell lines. In the case of organs, there was an increase in the expression of p38, especially p38b fraction in bone marrow, p38g and p38d fractions in the thymus, and p38b and p38d fractions in the spleen. A decrease in ERK-1 and p38d expression was noted in the spleen. In the case of cell lines, increases and decreases in MAPK expression were also investigated. In the Jurkat cell line, an increase was observed with ERK-2 and p38a. There was an increase in p38 in the WEHI 231 line, especially in the p38g fraction where it was over 47-fold. In the HL-60 line, TFX caused a significant decrease in ERK-2, p38d and JNK expression. In the J744E line the changes mainly concerned p38, there was an increase in the p38g fraction and a decrease in p38d.
In conclusion, Polish researchers confirmed the known effects of TFX and described its previously unknown effects, which will allow a better understanding of the experimental models and clinical effects.
The article will be published in the Polish journal “Journal of Biomedical Research and Therapeutics”, which can be read at: https://www.jbrt.pl