[ANTI-Hsp60 ANTIBODIES IN ARTERIAL HYPERTENTION DIFERENT DEGREE OF SEVERITY].

More than 12.1 million people with hypertension (32.2% of the adult population) were registered in Ukraine according to the official statistics on 1 January 2011. The etiopathogenesis of AH is not fully established. Hsp60 is the molecular chaperon/chaperonin, and it's expression significantly increases in response to different kinds of stress (emotional stress, infections, smoking etc). Elevated blood pressure is a mechanical stress to the endothelium and it can induce expression of heat-shock protein 60 (Hsp60) on the endothelial cell surface. Endothelial cells in the vessel wall can be damaged by (auto) immune reactions to Hsp60 present on the cell surface. Elevation of anti-Hsp60 in the circulation is associated with the presence and severity of coronary heart disease, atherosclerosis development, pathological changes in the small vessels of the brain etc etc. Specificity of the anti-Hsp60 antibodies and their role in the pathogenesis of AH has not been established. The aim of this work was to identify the level of anti-Hsp60 antibodies in the sera of patients with AH. 128 patients with AH were examined. To define level of anti-Hsp60 antibodies the sera 39 patients with AH, including 12 clinically healthy individuals (the family history are included the AH cases)--1 group, 19 patients with stage 2--2 group and 8 patients with stage 3--3 group were examined. The control group included 112 blood donors. Anti-Hsp60 antibodies in sera were determined by ELISA and immunobloting (Western-blotting). Recombinant piotein GroEL Escherihia coli (prokaryotic homologue of human Hsp60) and human Hsp60 were used as antigens. Average of levels of antibodies against GroEL and human Hsp60 in the serum of all groups twice exeeded the value of the control (P < 0.001). Antibodies to prokaryotic Hsp60 were prevailed in patients with AH. The seropositive serum to Hsp60 were detectived in patients, that had the risk of the AH complications by ELISA and immunoblotting. In addition, highly reactive IgG anti-Hsp60 antibodies purified by affinity chromatography from human sera of patients with AH recognized GroEL and human Hsp60 in immunoblotting. Elevated levels of anti-Hsp60 antibody in sera of patients with AH stage 3 correlated with pronounced changes in the target organs such as a massive recurrent hemorrhage into the retina, acute ischemic stroke, cardiosclerosis and angionephrosclerosis. It may indicate the involvement of anti-Hsp60 antibodies in the development of the target organ damage.

[DOXORUBICIN-INDUCED ALTERATIONS IN PRO-AND ANTIOXIDANT BALANCE AND THEIR CORRECTION BY CURCUMIN IN THE NEONATAL RAT CARDIOMYOCYTES CULTURE].

It was studied the effect of doxorubicin on the HIF system and the pro-antioxidant balance of neonatal cardiomyocytes as well as the possibility of the oxidative stress correcting using curcumin. It has been revealed that the expression of mRNA HIF-1α using doxorubicin at a dose of 0.5 µM was 2.9 ± 0.8 cu, so it decreased by 20% compared to control--3.6 ± 0.7 cu (P < 0.05). The level of expression of the HIF target gene PDK-1 also significantly decreased (4 times). During the incubation with doxorubicin, the number of live cells decreased by 50.4% relative to control. And after using doxorubicin and curcumin together, the percentage of dead cells decreased by 7,7 compared to doxorubicin only. Doxorubicin intoxication led to a significant increase in the secondary products of lipid peroxidation (TBARS) in cardiomyocytes by 3.6 times and hydrogen peroxide by 64%. Prolonged incubation with doxorubicin reduced the enzymatic activity of Mn-SOD by 32%, while catalase activity increased by 72% compared to control. Adding of curcumin to the cardiomyocyte cell culture contributed to increasing of the Mn-SOD activity by 14%, catalase--by 23%. The level of TBARS increased by 1,4 times compared with the control, and the level of H2O2 increased by 20%. The joint use of doxorubicin and curcumin resulted in a significant reduction of free radical oxidation unlike effect of doxorubicin per se. Specifically, there was lessening of TBARS and H2O2 (at 56.7 and 18.4% respectively), while decreasing of the catalase hyperactivation (19%) and rising of the Mn-SOD activity (35%).

Comparative characteristic of thiamine antagonists on apoptosis induction in different types of nerve cell lines.

Abnormalities in oxidative metabolism and inflammation accompany many neurodegenerative diseases. The mechanisms of neurodegeneration induced by thiamine deficiency remain incompletely elucidated. The susceptibility of various types of nerve cells to thiamine (vitamin B) antagonists--oxythiamine (OT), pyrithiamine (PT) and amprolium (Am) was investigated. Four cell lines (neuronally differentiated rat PC-12, rat astrocytes DITNC, neuronally differentiated human SH-SY5Y and human astrocytic cells 1321N1) were used for experiments as neural cell models. When different cell types were cultivated with thiamine antagonists, a significant decrease of viability was detected in a time- and dose-dependent manner as demonstrated by the WST-1 colorimetric assay. These data were similar to those of caspase 3 activity and DNA fragmentation induced by thiamine antagonists. All tested cell lines were more vulnerable to OT and PT than to Am. Am displayed a pronounced damaging action on neuronal cells and had a modest influence on astrocytes. The last observation gives the basis to suppose, that neuronal cells need external arrival of thiamine more than astrocytes. Thus, the results testify that various types of nerve cells have different susceptibility to the thiamine antagonists and this relates to extent of apoptosis development.