Background The disordered implementation of proliferation and apoptosis processes in the cells of monocyte-phagocyte system (MPS) is significant in the complex of pathogenetic events characteristic for rheumatoid arthritis (RA). In particular, the characteristic for RA proliferation of the synovial tissue cells and development of pannus are related to low apoptotic activity of these cells. Since the induction of apoptosis in immune competent cells is considered as one of the approaches to arrest the disorders in immune system the new strategy of RA treatment may be the correction of apoptotic processes in MPS cells using the ones of fetal liver (FLCs) .
Objectives The research aim was to comparatively assess the development of apoptotic processes in MPS cells (peritoneal macrophages) of the animals with adjuvant arthritis (AA) prior to and after their treatment with cryopreserved FLCs (cFLCs).
Methods The researches were performed in experimental model of human rheumatoid arthritis (RA), AA induced in mice by means of subplantar introduction of Freund’s complete adjuvant . FLCs were intravenously injected in a dose of 5’106cells/mouse to the 7th and 14th days of AA development. FLCs were cryopreserved under 5% DMSO protection according to 4-step program. As a positive control there was used the treatment of the animals with dexamethasone. The number of dead MPS cells was evaluated using propidium iodide by flow cytometry. Expression of CD95 cascade proteins in lysate of MPS cells was analyzed by western blotting. Catalytic activity of caspases-3 and -8 in MPS cells was found on the results of measuring proteolytic cleavage of fluorogenic ZIETD-AFC or AC-DEVD-AMC substrates.
Results It has been shown that during AA development the MPS cells manifest the resistance to apoptosis. There was found pro-apoptotic effect of FLCs after their introduction to the recipients with AA. In addition there have been shown significant differences in the mechanisms of implementation of FLCs and glucocorticoids (GCs) pro-apoptotic activity. After introduction of FLCs there were reduction of FLIP protein production and activation of caspase-8 in MPS cells, whilst after application of GC no similar effect was observed. There was demonstrated the dependence of manifesting the correcting effect of FLCs on the type of introduced material and the terms of their introduction. Maximal pronounced pro-apoptotic effect was found when introducing cFLCs to the 7th day of AA which is confirmed with the highest indices of the activity of caspases- 3 and -8 in the cells of MPS of the recipients with AA.
Positive effect of the introduced cFLCs to the 7th day of AA has been established to be kept for two weeks after treatment, emphasizing the value of FLCs application protocol at the given pathology.
Conclusions More manifested pro-apoptotic effect of cFLCs if compared with native FLCs may be stipulated by the activation under the effect of cryopreservation factors of the production of different death receptor ligands.
Yampolskaya E.E., Shatneva O.M., Bondarovich N.A., Goltsev A.N. Apoptotic mechanisms in monocytic-phagocytic system during rheumatoid arthritis development after fetal liver cells introduction // Biotechnology. – 2012. – Vol.5, N5. – P. 82-90.
Mischenko O.Ya., Kotvitska A.A. Pharmacological efficiency of analbene emulsion in the model of adjuvant arthritis in rats // Visnyk Farmatsii. – 2001. – N3. – P. 124-125.
Disclosure of Interest None Declared