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ФГБОУ ВО Тихоокеанский государственный медицинский университет Минздрава России. Владивосток, Россия Цель. Определить уровень экспрессии эндотелиальных факторов (VEGF-A, CD34 +) в тканях суставов при экспериментальной артериальной гипертонии (АГ), гиперлипидемии (ГЛ) и их сочетании. Материал и методы. Экспериментальное исследование проведено на 24 половозрелых самцах беспородных морских свинок. Все животные были разделены на 4 группы по 6 особей в каждой. В первой группе воспроизводили АГ путем внутримышечных инъекций раствора гидрокартизона. ГЛ у второй группы моделировали при внутрибрюшинной инъекции 1 раз в 3 нед. раствора Твин-80 и модификации рациона. В третьей группе воспроизводили сочетание АГ и ГЛ путем инъекций раствора гидрокартизона, Твин-80 и модификации рациона питания. На 60 сут. животные были выведены из эксперимента эвтаназией. Произведен забор тканей скакательных суставов задних конечностей, в которых с помощью пероксидазной иммуногистохимической реакции определяли экспрессию эндотелиального фактора роста сосудов A (VEGF-A), CD34. Результаты. Установлено, что при экспериментальной АГ в субхондральной кости наблюдается повышенная экспрессия VEGF-A. При экспериментальной ГЛ в суставном хряще и субхондральной кости наблюдается преимущественная индукция VEGF-A и CD34. При комбинированном воздействии сердечно-сосудистых факторов наблюдается наиболее высокая экспрессия молекулярных механизмов кардиометаболического стресса, включая VEGF-A и CD34. Заключение. Получены результаты, свидетельствующие о том, что сердечно-сосудистые факторы оказывают влияние на суставные ткани. Это позволяет сделать предположение о принадлежности тканей суставов к органам-мишеням сердечно-сосудистых заболеваний и позволяют судить о важной роли сердечно-сосудистой коморбидности в патогенезе дегенеративно-воспалительных заболеваний суставов. Ключевые слова: остеоартрит, артериальная гипертония, гиперлипидемия, суставной хрящ, субхондральная кость. Конфликт интересов: не заявлен.

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Aim. To assess the relationship between textural characteristics of the subchondral bone and standard X-ray data, to determine markers of subchondral bone remodeling in gonarthrosis.Methods. The studied group included 92 patients aged 66.1±10.5 years with I-IV grades osteoarthritis by the Kellgren, in the comparison group - 24 volunteers aged 29.6±5.96 years without clinical or radiological signs of gonarthrosis. Standard digital X-ray of the knee joint was performed. On the image, the area of interest was chosen, including a portion of the subchondral bone of 48±2×90±4 pixels of size. According to the area texture, the gray-level histogram and 3D graph of the pixels intensity distribution in area were made.Results. The distribution of individual pixel values relating to the average gray-level values showed an inverse correlation with the disease stage (r=-0.52, p=0.00004) and the presence of large osteophytes (r=-0.40, p=0.002). Extremum of 3D histogram minimum value directly correlated with radiographic stage of gonarthrosis (r=0.42, p=0.0009), patients’ age (r=0.33, p=0.01) and the osteophytes number (r=0.43, p=0.0007). This figure was higher in the group of patients with osteoarthritis (p=0.009) and significantly decreased with the disease progression (p=0.04).Conclusion. For the first time the analysis of 3D surface reconstruction depending on the gray-level pixel values was used, which showed good characteristics on the distinguishing groups of patients with osteoarthritis, and comparability with standard radiographic protocol data; the best results demonstrated the minimum value at 3D histogram that had significant variation depending on the disease stage.

The aim of this study was to analyze the anti-inflammatory effect of VA692, in comparison with celecoxib. In addition, we would establish if the new compound presents any advantages over other COX-2 inhibitors already available in therapeutic use. By iTRAQ methodology, we quantitatively analyzed the different expressed profiles in T/C-28a2 cell line treated with the studied drugs in presence of IL-1b. Methods: Human T/C-28a2 chondrocytes cell line were generated by Goldring group. Human articular cartilage was obtained from femoral heads of five OA patients. Cells were incubated with VA692 and celecoxib (1, 0.5 and 0.2mM) with interleukin (IL)-1b (5ng/ml) for 48hrs. The expression of inflammatory cytokines and anti-oxidant enzymes was evaluated by quantitative qRT-PCR, prostaglandins (PG)-E 2 release by ELISA. Apoptosis levels and ROS production were evaluated by flow cytometry. Proteins extracted by T/C-28a2 cell line was employed to carry out western blot analysis and the iTRAQ methodology. Statistical analysis was performed by ANOVA and Bonferroni multiple comparison tests. Results: IL-1b-stimulated chondrocytes showed a significant increase (p<0.001) of COX-2, IL-1b, IL-6, IL-8, superoxide dismutase (SOD)-2 and catalase (CAT) gene expression, as well as of PGE 2 levels in OA chondrocyte and in T/C-28a2 cell line. The tested drugs significantly counteracted the effect of IL-1b, with a better modulation by VA692 1mM in T/C-28a2 (p<0.01 for COX-2, IL-1b, IL-8, CAT; p<0.001 for IL-6, SOD-2 and PGE 2). Regarding apoptosis and mitochondrial superoxide anion production, the new drug was able to significantly reduce (p<0.05) their increase induced by IL-1b (p<0.05). Proteomic analysis led to the identification of 797 proteins in T/C-28a2 cell line, 123 of which were significantly modulated by VA692 in presence of IL-1b (p<0.001), while 34 were modulated by IL-1b alone (p<0.05). 21 proteins were commonly modulated by the two groups, indicating that 101 proteins were regulated by VA692 in a specific manner. Among the proteins downregulated by VA692, some with structural function were detected, responsible for cytoskeleton reorganization, as well as chaperones (heat shock proteins) and glycolitic enzymes. Proteins involved in calcium metabolism and in ribosome biogenesis resulted up-regulated instead, as well as SOD-2 as confirmed by western blot analysis. Conclusions: Our data demonstrated the anti-inflammatory effect of VA692, suggesting also its anti-apoptotic and anti-oxidant role. The proteomic analysis demonstrated that VA692 induced not only an antiinflammatory regulation in chondrocytes but, interestingly, seemed to regulate their anabolic response. On the basis of our results, we can hypothesize that VA692 could present any advantages over other COX-2 inhibitors already available for therapeutic use. However, further in vitro and in vivo experiments are necessary to elucidate and confirm the importance of this pharmacological compound before its use in the therapeutic approach of joint diseases.

The objective of the study was to assess articular cartilage (AC) and subchondral bone (SCB) remodeling, expression of matrix metalloproteinase 9 (MMP-9) in tissues, vascular endothelial growth factor (VEGF) in experimental osteoarthritis (OA). Material and methods Experimental comparative study was conducted on 12 outbred guinea pigs of both sexes aged 28-30 weeks that were divided into 2 groups of 6 animals each. An injury to the knee joints of hind limbs of control and experimental animals was mechanically simulated by closed scarification using a sterile needle. No treatment was provided for controls. Experimental animals were given an intraarticular injection of betamethasone (BMZ) of 0.1 mg/kg every two weeks after two weeks of injury. Two subjects of each group were euthanized at 30, 45, 60 days and knee samples collected. Immunohistochemical expression of VEGF and MMP-9 was determined in tissues. Results A statistically significant decrease in VEGF positive chondrocytes and precipitate density, an increase in positive chondrocytes and intensity of tissue response to MMP-9 as compared to those in controls was observed in BMZ animals at 30 days of experiment. A statistically significant decrease in VEGF positive cells and precipitate density, an increase in VEGF positive chondrocytes as compared to those in controls were observed in BMZ animals at 45 days of experiment. Significant reduction in VEGF positive cells and deposit density, significantly higher density of MMP-9 positive precipitates as compared to those in controls were noted at 60 days of BMZ injections. Conclusion Intraarticular injections of BMZ demonstrated a negative effect on AC and SCB with articular tissue remodeling initiated through activation mechanisms of extracellular matrix degradation, as evidenced by high expression of MMP-9. BMZ was shown to block pathological angiogenesis via VEGF inhibition.

Тихоокеанский государственный медицинский университет, г. Владивосток, Россия РефератЦель. Установить клинико-патогенетические закономерности между уровнями апоптоза и регуляторами роста и дифференцировки (ингибитором роста 1, индуцированным оксидативным стрессом; фактором роста и диф-ференцировки 5) при остеоартрите. Методы. В условиях ревматологического кабинета Владивостокской поликлиники №3 были обследованы 65 па-циентов с остеоартритом коленных суставов I-IV стадий по Kellgren в возрасте 66,5±8,0 лет. В качестве группы сравнения в исследование были включены 25 добровольцев, сопоставимых с основной группой по полу и воз-расту, не имевших клинических и рентгенологических проявлений остеоартрита. Для определения концентра-ций искомых молекул в крови пациентов, включённых исследование, использовали иммуноферментный анализ. Результаты. У больных остеоартритом по сравнению с группой контроля отмечены статистически значимые повышения уровней Fas, фактора роста и дифференцировки 5 и соотношения «фактор роста и дифференциров-ки 5/ингибитор роста 1, индуцированный оксидативным стрессом». Показатели Fas были достоверно ниже на «поздних» III-IV стадиях остеоартрита по сравнению с I и II стадиями. Уровень фактора роста и дифференци-ровки 5 имел более низкие значения у больных с III-IV стадиями остеоартрита по сравнению с I и II. По мере прогрессирования рентгенологических симптомов остеоартрита зарегистрировано снижение соотношения «фактор роста и дифференцировки 5/ингибитор роста 1, индуцированный оксидативным стрессом», которое было достоверно ниже на II и III стадиях по сравнению с I стадией. Вывод. Внешний путь апоптоза имеет большое значение при формировании болевого синдрома при остеоар-трите, а его поддержание реализуется по другим механизмам, к которым можно отнести влияние оксидатив-ного стресса через опосредованное ингибитором роста 1, индуцированным оксидативным стрессом, угнетение клеточного цикла, снижение участия фактора роста и дифференцировки 5 в процессах дифференцировки и ре-гуляции синтеза белков внеклеточного матрикса хрящевой ткани. Ключевые слова: остеоартрит, апоптоз, клеточная дифференцировка, оксидативный стресс. Clinical and pathogenetic interrelation between molecular regulation of apoptosis and cell differentiation in osteoarthritis M.A. Kabalyk, N.G. Plekhova, A.V. Lagureva, A.B. Sunyaykin Pacific State Medical University, Vladivostok, RussiaAim. To determine clinical and pathogenetic relationship between the levels of apoptosis and growth and differentiation regulation (growth inhibitor 1 induced by oxidative stress, growth/differentiation factor 5) in osteoarthritis. Methods. In a rheumatology office of Vladivostok polyclinic №3 65 patients with knee osteoarthritis Kellgren grade 1-4 aged 66.5±8.0 years were examined. 25 healthy volunteers matched by sex and age without clinical and radiologic manifestations of osteoarthritis were included into control group. To measure concentration of the studied molecules in study patients' blood, ELISA method was used. Results. Patients with osteoarthr...

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