Tag Archives: rheumatoid arthritis
Characteristics of Ca(2+), Mg(2+)-ATPases of peripheral blood lymphocytes of patients with rheumatic pathology
R. V. Fafula, U. P. Efremova, Z. D. Vorobets
Danylo Halytski Lviv National Medical University, Ukraine;
e-mail: roman_fafula@ukr.net, vorobets@meduniv.lviv.ua
The analysis of the kinetic properties of Ca2+, Mg2+-ATPase of saponin-perforated peripheral blood lymphocytes of donors and patients with rheumatoid arthritis and ankylosing spondylitis was carried out. When analyzing the alterations in hydrolase activity of Ca2+, Mg2+-ATPase it was shown that affinity of Ca2+, Mg2+-ATPase of plasma membrane and membranes of endoplasmic reticulum for ATP do not significantly differ. It was found that the inhibition of examined enzyme systems occurs by mixed type both due to the reduction of maximum reaction rate and to the decrease of Ca2+, Mg2+-ATPase affinity for ATP in conditions of rheumatic pathology in the immunocompetent cells. It was identified that Ca2+, Mg2+-ATPase had significantly lower affinity for Ca2+ in lymphocytes of persons with rheumatic disorders than in donors.
Kinetic properties of Na(+), K(+)-activated, Mg(2+)-dependent ATP-hydrolysis of blood lymphocytes in patients with rheumatoid arthritis and ankylosing spondyloarthritis
R. V. Fafula, U. P. Efremova, N. E. Lychkovska, Z. D. Vorobets
Danylo Halytski Lviv National Medical University, Ukraine;
e-mail: roman_fafula@ukr.net; vorobets@meduniv.lviv.ua
The comparative analysis of the kinetic properties of ouabain-sensitive Na+, K+-ATPase activity of saponin-perforated blood lymphocytes of donors and patients with rheumatoid arthritis (RA) and ankylosing spondyloarthritis (AS) was carried out. When analyzing the alterations in hydrolase activity of the examined enzyme it was shown that in the blood lymphocytes of patients with RA and AS the primary active transport of Na+ and K+ ions is less intensive in comparison with practically healthy donors, but it is characterized by almost the same capacity as in donors. The affinity constant of Na+, K+-ATPase for ATP in the blood lymphocytes in patients with RA and AS is greater 3.1 and 2.5 times, respectively, in comparison with healthy donor. It was found that in conditions of rheumatic pathology in immunocompetent cells the inhibition of Na+, K+-ATPase activity is not related to the reduction of maximum reaction rate, but is related to the decrease of Na+, K+-ATPase affinity to ATP. However, Mg2+-binding center of Na+, K+-ATPase in patients with RA and AS remains native. It was identified that the affinity constant of Na+, K+-ATPase to Na+ ions in the blood lymphocytes of patients with RA and AS is 2.75 times lower than its value in healthy donors. Na+, K+-ATPase of the blood lymphocytes of patients with RA and AS retains its native receptor properties and sensitivity to ouabain does not change.
Proteolytic activity of IgGs from blood serum of wistar rats at experimental rheumatoid arthritis
Yu. Ya. Kit1, S. L. Myronovsky3, I. I. Kril’2,
A. M. Havrylyuk2, V. V. Chop’yak2, R. S. Stoika1
1Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
2Danylo Halytsky Lviv National Medical University, Ukraine;
3Ivan Franko National University of Lviv, Ukraine;
e-mail: kit@cellbiol.lviv.ua
The aim of this work was to study the proteolytic activity of IgGs purified from blood serum of Wistar rats at experimental rheumatoid arthritis (ERA) induced by an injection of bovine collagen of type II. Twenty rats were immunized with a preparation of bovine collagen II (Sigma-Aldrich, USA) in the presence of complete Freund’s adjuvant. ERA development was determined by inflammation in limbs of treated animals. IgG preparations were isolated from blood serum of immunized and non-immunized animals by precipitation of antibodies with 33% ammonium sulfate followed by chromatography on the Protein G-Sepharose column. Human histone H1, bovine collagen II, calf thymus histones, myelin basic protein (MBP), bovine serum albumin (BSA), and bovine casein were used as substrates of the proteolytic activity of IgGs. It was found that IgG preparations from blood serum of rats with ERA were capable of cleaving histone H1 and MBP, however, they were catalytically inactive towards collagen II, casein, BSA, and core histones. IgGs from blood serum of non-immunized rats were proteolytically inactive towards all used protein substrates. Thus, we demonstrated that immunization of rats with bovine collagen II induced IgG-antibodies possessing the proteolytic activity towards histone H1 and MBP. This activity might be associated with the development of inflammatory processes in the immunized rats.