scholarly journals POS0169 FETUIN-A AS A MARKER OF OSTEOPOROSIS AND OSTEOPOROTIC FRACTURES IN PATIENTS WITH RHEUMATOID ARTHRITIS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 297.2-297
Author(s):  
Y. Akhverdyan ◽  
E. Papichev ◽  
В. Zavodovsky ◽  
L. Seewordova ◽  
J. Polyakova
Keyword(s):  
Rheumatoid Arthritis ◽  
Blood Serum ◽  
Bone Mineralization ◽  
Expectant Management ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Individuals ◽  
High Concentration ◽  
Mineral Density ◽  
Fetuin A ◽  
Apparently Healthy

Background:The main mechanism of the effect of fetuin-A (FeA) on bone metabolism is its ability to bind calcium and proteins of the TGF-β family. It has been proven that the optimal concentration of TGF-β is necessary for the differentiation of bone tissue, and a high concentration inhibits bone mineralization. Thus, adequate osteogenesis is based on a complex balance between FeA and TGF-β levels. It can be assumed that the determination of the FeA level in the blood of patients with rheumatoid arthritis (RA) will help to optimize the diagnosis and predict the severity of osteoporosis (OP).Objectives:to study the possibility of predicting the development of osteoporosis and osteoporetic fractures in patients with RA, depending on the level of FeA in blood serum.Methods:We examined two groups of patients (52 patients with RA complicated by OP, 58 patients with RA without OP) and 30 apparently healthy individuals. The age of the surveyed ranged from 18 to 72 years, the average duration of the disease was 7.53±0.89 years. In both groups, the FeA level was determined by an indirect enzyme-linked immunosorbent assay using a commercial test. Bone mineral density (BMD) was also measured in both groups (Lunar DPX-NT GE).Results:The average FeA level in the group of RA patients was lower than in the group of conventionally healthy individuals (731.21±109.9 μg/ml and 812.9±76.2 μg/ml, respectively; F=13.34; p=0,0004). The normal FeA level was calculated using the formula M±2σ in the group of apparently healthy individuals and ranged from 653.55 μg/ml to 972.19 μg/ml.A decreased level of FeA was found in 20 patients (86.96%) in the group of patients with OP and only in 3 (13.04%) patients with RA who did not suffer from OP (p<0.001). It can be concluded that patients with RA and a low concentration of FeA in the blood serum have a higher risk of developing OP.In the group of patients with normal FeA level, osteoporetic fractures were observed in 12 (13.79%) patients and were absent in 75 (86.21%) patients (p<0.001). Thus, RA patients with normal serum FeA levels have a lower risk of osteoporetic fractures.We also found a positive significant correlation between the level of FeA and BMD in the femoral neck area. In the group of patients with a reduced FeA level (23 people), the mean BMD values were 0.732±0.022 g/cm2, and in the group of patients with a normal FeA level (87 patients) - 0.890±0.014 g/cm2 (p<0.001, F=27.663). The obtained values are in agreement with the literature data on the effect of the serum FeA concentration on the BMD values.Conclusion:We consider it expedient to determine the serum FeA concentration in patients with RA. At a FeA level of 653.55 μg/ml and below, a higher risk of developing OP and osteoporetic fractures can be predicted. In this case, the patient is shown a standard examination for osteoporosis. At values of 653.55 μg/ml and above, a more expectant management of the patient is allowed. Thus, by determining the serum concentration of FeA, it is possible to implement an integrated approach to the patient and to optimize the schemes for the diagnosis of OP in patients with RA.Disclosure of Interests:None declared

scholarly journals POS0451 SERUM FETUIN-A AND VISFATIN LEVELS IN WOMEN WITH RHEUMATOID ARTHRITIS DEPENDING ON DISEASE ACTIVITY

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 455.2-456
Author(s):  
Y. Akhverdyan ◽  
В. Zavodovsky ◽  
E. Papichev ◽  
J. Polyakova ◽  
L. Seewordova
Keyword(s):  
Rheumatoid Arthritis ◽  
Blood Serum ◽  
Disease Activity ◽  
Transforming Growth Factor ◽  
Average Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Healthy Individuals ◽  
Fetuin A ◽  
Grade Ii

Background:In recent years, the systemic effects of a number of cytokines have been actively studied, in particular, fetuin-A is considered a negative protein of the acute phase response, and visfatin, on the contrary, affects the activation of the cytokine cascade and has a pro-inflammatory effect. Taking into account that women suffer from rheumatoid arthritis (RA) more often, we investigated the levels of fetuin-A and visfatin in the blood serum of females in comparison with a group of healthy individuals and depending on the activity of the disease.Objectives:to study the levels of fetuin-A and visfatin in the blood serum of women suffering from RA, depending on the activity of the diseaseMethods:The study included 110 women with RA and 30 apparently healthy individuals. The inclusion criteria were: a diagnosis of RA verified based on the criteria of the American College of Rheumatology/European Anti-Rheumatic League (ACR/EULAR) 2010. The patients’ age ranged from 18 to 90 years. The control group included 30 conventionally healthy individuals. Serum fetuin-A and visfatin levels were determined by indirect enzyme-linked immunosorbent assay using commercial kits. RA activity was determined by the DAS28-CRP index. Activity 0-I was in 33 (30%) patients, grade II in 67 (60.9%), grade III in 10 (9.09%) patients.Results:The normal level of fetuin-A was calculated using the formula M±2σ in the group of conventionally healthy individuals and ranged from 653.55 to 972.19 μg/ml. In patients with grade 0-I RA activity according to DAS28, the mean serum fetuin-A level was 843.92±130.73 μg/ml, in patients with grade II activity - 742.37±98.85 μg / ml, with III the degree of activity - 663.9±123.7 μg/ml (p<0.001).The average level of visfatin in the blood serum in healthy individuals was 2.43±0.17 ng/ml. The level of normal values of visfatin in healthy individuals, defined as M±2σ, ranged from 0 to 5.07 ng/ml. The average level of visfatin in patients with RA was 6.27±0.18 ng/ml, which is significantly higher than in healthy individuals (p<0.001).In patients with 0-I degree of RA activity according to DAS28, the average level of visfatin in blood serum was 4.94±0.02 ng/ml, in patients with degree II activity - 5.08±0.02 ng/ml, with III degree of activity - 6.82±0.23 ng/ml (p<0.001).Conclusion:Thus, the level of fetuin-A in the blood serum of patients with RA is significantly lower in the case of a high degree of disease activity. The level of visfatin in the blood serum in women with RA is significantly higher in patients with a higher degree of disease activity. Therefore, the concentration values of fetuin-A and visfatin in the blood serum of patients with RA can be used in an integrated assessment of the prognosis of disease activity.References:[1]Inoue K, Ikeda Y, Yamanaka S, et al. Serum fetuinA levels in patients with rheumatoid arthritis [abstract]. Atherosclerosis. 2002;9(Suppl 1):233. doi: 10-1016/s1567-5688(08)70930-9[2]Janssens K, ten Dijke P, Janssens S, et al. Transforming growth factor-beta1 to the bone. Endocrine Reviews. 2005;26(6):743-74. doi:10.1210/er.2004-0001[3]Polyakova J, Korolik O, Papichev E, et al. The role of «new» cytokines in the pathogenesis rheumatoid arthritis. Ann Rheum Dis. 2018; 78(2): 1497Disclosure of Interests:None declared

scholarly journals AB0103 THE AMBIGUOUS ROLE OF TISSUE CYTOKINES IN THE PATHOGENESIS OF RHEUMATOID ARTHRITIS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1350.1-1351
Author(s):  
O. Korolik ◽  
В. Zavodovsky ◽  
E. Papichev ◽  
Y. Polyakova ◽  
S. L ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
High Activity ◽  
Inflammatory Cytokines ◽  
Stage Iii ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Individuals ◽  
Fetuin A ◽  
High Level ◽  
Extraarticular Manifestations ◽  
Pro Inflammatory Cytokines

Background:Cytokines stimulate the inflammatory response in the synovial membrane with rheumatoid arthritis (RA), initiate apoptosis of chondrocytes, activation of osteoclasts. The progression of comorbid diseases is also associated with the influence of cytokines. At the same time, anti-inflammatory cytokines are produced in various tissues. Their role in the pathogenesis of RA and its complications is ambiguous.Adiponectin (A) and Fetuin A (FA) are classified as negative acute phase proteins. Their concentration decreases with an increase in the level of pro-inflammatory cytokines: TNF-α, IL-1 and IL-6. Molecules A and FA, regardless of various factors and from each other, have similar effects in relation to pro-inflammatory cytokines, lipid and carbohydrate metabolism.Visfatin (V) and Nesfatin-1 (N-1) are pro-inflammatory adipokines. B is produced by cells of the mononuclear phagocytic system and connective tissue. N-1 - is produced by the cells of the intermediate and medulla oblongata and by the cells of the gastric mucosa.Objectives:to study the correlation of B, H-1, A and FA with the severity of inflammation in RAMethods:60 patients with RA and 30 healthy individuals were examined. The level of cytokines was determined by an indirect enzyme-linked immunosorbent assay using commercial test systems (Bio Vendor, cat No. RD195023100, Bio Vendor Human Fetuin-A, RaiBiotech, cat No. EIA-VIS-1, RaiBiotech, cat No. EIA-NESF). All patients underwent a full examination. Diagnosed with 2010 EULAR / ACR recommendations.Results:A decreased level of A (less than 0.8 μg/ml) was detected in 15 patients (25%), F-A (less than 653.55 μg/ml) in 16 (27%), a high level of V (more than 39 ng/ml) - in 55 (91%), N-1 (more than 37.95 ng/ml) - in 36 (60%), which is significantly more often than in healthy individuals. No significant difference in the levels of determined adipokines was found depending on the gender and body weight of patients with RA. The level of cytokines in RA is associated with high activity according to DAS 28, positivity by Anti-CCP, extraarticular manifestations of RA. The greatest correlation with extraarticular manifestations is with cutaneous and cerebral vasculitis. The levels of FA and N-1 also correlated with more pronounced radiological changes (X-ray stage III). FA circulating inhibitor of ectopic calcification. N-1 level is positively correlated with systolic blood pressure.Conclusion:A low level of A and FA, a high level of V and N-1 is characteristic of RA with the presence of high activity and positivity in the RF and Anti-CCP. An increased level of B is determined by more than 90% of patients, which indicates its high pro-inflammatory activity. The level of F and N-1 is also associated with the degree of damage to bone tissue (stage III, a lot of erosion). A positive correlation of level V and N-1, negative A and FA with the severity of inflammation in RA confirms the involvement of these proteins in the pathogenesis. A high level of A and V increases the risk of developing cardiovascular diseases and their complications, the effect of N-1 and FA is being studied. The effect of cytokines on osteoclasts and osteoblasts in RA is ambiguousReferences:[1]Visfatin and Rheumatoid Arthritis: Pathogenetic Implications and Clinical Utility. Polyakova Y. Curr Rheumatol Rev.2019[2]Serum nesfatin -1 as a marker of systemic inflammation in rheumatoid arthritis. Kvlividze T. Klinicheskaya Laboratornaya Diagnostika.2019; 64 (1):53-56 (in Russ)[3]Fetuin-A. Novel hepatokine in rheumatoid arthritis laboratory diagnostics. Papichev E. Klinicheskaya Laboratornaya Diagnostika.2018; 63 (12):756-760 (in Russ)Disclosure of Interests:None declared

scholarly journals AB0827 SERUM NESPHATIN-1 IN PATHOGENESIS RHEUMATOID ARTHRITIS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 1437.2-1438
Author(s):  
T. Kvlividze ◽  
V. Polyakov ◽  
В. Zavodovsky ◽  
Y. Polyakova ◽  
L. Seewordova ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Blood Serum ◽  
Inflammatory Cytokines ◽  
Inflammatory Markers ◽  
Healthy People ◽  
Healthy Individuals ◽  
Markers Of Inflammation ◽  
High Level ◽  
Pro Inflammatory Cytokines

Background:Interest in highly specialized tissue cytokines contributed to the discovery of new biologically active molecules. Nesfatin-1 (NF) - discovered in 2006 as an anorexigenic factor. NF-1 is believed to be involved in the regulation of energy homeostasis by regulating appetite and water intake. The role of NF-1 in the pathogenesis of inflammatory diseases is poorly understood. Recently, studies have found a relationship between an increased level of NF-1 and inflammatory markers in various pathologies.Objectives:Study of the level of nesfatin-1 in the blood serum of healthy people, determination of the correlation between the level of NF-1 with the severity of clinical symptoms and classic markers of inflammation in patients with RA.Methods:120 persons were examined: 90 patients with RA and 30 healthy people. All patients underwent a complete clinical and laboratory examination. Plasma NF-1 levels were determined using commercial test systems (RaiBiotech, cat # EIA-NESF) according to the manufacturer’s instructions. Patients with various forms of RA were comparable in age to the group of healthy individuals. Statistical processing of clinical examination data was carried out using the “STATISTICA 10.0 for Windows” software package. Quantitative data were processed statistically using the parametric Student’s t-test, qualitative data using the non-parametric chi-square test. The significance of differences between groups was determined using analysis of variance. The results were considered statistically significant at p <0.05.Results:The average level of NF-1 in blood serum in healthy individuals was 31.79 ± 3.21 ng / ml (M ± σ). The level of normal NF-1 values in healthy individuals, defined as M ± 2σ, ranged from 25.3 to 37.83 ng / ml. There was no significant difference in the levels of circulating NF-1 and BMI in healthy individuals and patients with RA (p> 0.05). The inverse relationship of a lower level of NF-1 with an increase in BMI was not significant.Group 1 (66 patients with RA) with increased serum NF-1 levels (> 37.83 ng / ml), and group 2 (44 patients) with normal values (<37.83 ng / ml). A high level of NF-1 was characteristic for patients with high activity according to DAS28, RF seropositive, ACCP-positive, with extra-articular manifestations, who had been ill for 10 years or more. A reliable relationship between the level of NF-1 in the blood serum and laboratory parameters of RA activity - ESR, CRP, was shown, and secondary synovitis was more common. Our data show a direct correlation between the NF-1 level of the pro-inflammatory markers of RA.Conclusion:The positive correlation between the level of NF-1 and classical markers of inflammation, such as CRP and ESR, confirms the involvement of NF-1 in the pathophysiology of inflammation in RA. This is also evidenced by the correlation of a high level of NF-1 in the blood serum with a more severe clinical picture of RA. It is known that NF-1 can promote the release of pro-inflammatory cytokines such as interleukin-8 (IL-8), interleukin-6 (IL-6), and macrophage inflammatory protein-1a (MIP-1a) in the chondrocytes of RA patients.It is necessary to further study the role of NF-1 in the pathogenesis of systemic inflammatory reactions and the possibility of targeting pro-inflammatory cytokines, the possibility of regulating the level of NF-1 by drugs.References:[1]Kvlividze T.Z., Zavodovsky B.V., Akhverdyan Yu.R. Kvlividze T.Z., Zavodovsky B.V., Akhverdyan Yu.R., Polyakova Yu.V., Sivordova L.E., Yakovlev A.T., Zborovskaya I.A. Serum nesfatin -1 as a marker of systemic inflammation in rheumatoid arthritis. Klinicheskaya Laboratornaya Diagnostika (Russian Clinical Laboratory Diagnostics). 2019; 64 (1): 53-56 (in Russ.).Disclosure of Interests:None declared

scholarly journals Analytical Characterization of an Enzyme-Linked Immunosorbent Assay for the Measurement of Transforming Growth Factor β1 in Human Plasma

2018 ◽  
Vol 3 (2) ◽  
pp. 200-212 ◽  
Author(s):  
Brendan M Giles ◽  
Timothy T Underwood ◽  
Karim A Benhadji ◽  
Diana K S Nelson ◽  
Lisa M Grobeck ◽  
...  
Keyword(s):  
Growth Factor ◽  
Human Plasma ◽  
Patient Selection ◽  
Transforming Growth Factor ◽  
Sandwich Elisa ◽  
Transforming Growth Factor Β ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Individuals ◽  
Tgf Β1 ◽  
Apparently Healthy

Abstract Background The transforming growth factor β (TGF-β)–signaling pathway has emerged as a promising therapeutic target for many disease states including hepatocellular carcinoma (HCC). Because of the pleiotropic effects of this pathway, patient selection and monitoring may be important. TGF-β1 is the most prevalent isoform, and an assay to measure plasma levels of TGF-β1 would provide a rational biomarker to assist with patient selection. Therefore, the objective of this study was to analytically validate a colorimetric ELISA for the quantification of TGF-β1 in human plasma. Methods A colorimetric sandwich ELISA for TGF-β1 was analytically validated per Clinical and Laboratory Standards Institute protocols by assessment of precision, linearity, interfering substances, and stability. A reference range for plasma TGF-β1 was established for apparently healthy individuals and potential applicability was demonstrated in HCC patients. Results Precision was assessed for samples ranging from 633 to 10822 pg/mL, with total variance ranging from 28.4% to 7.2%. The assay was linear across the entire measuring range, and no interference of common blood components or similar molecules was observed. For apparently healthy individuals, the average TGF-β1 level was 1985 ± 1488 pg/mL compared to 4243 ± 2003 pg/mL for HCC patients. Additionally, the TGF-β1 level in plasma samples was demonstrated to be stable across all conditions tested, including multiple freeze–thaw cycles. Conclusions The ELISA described in this report is suitable for the quantification of TGF-β1 in human plasma and for investigational use in an approved clinical study.

scholarly journals Fetuin-a and secondary osteoporosis in patients with rheumatoid arthritis

2019 ◽  
Vol 27 (3) ◽  
pp. 360-366
Author(s):  
Evgeniy V. Papichev ◽  
Boris V. Zavodovsky ◽  
Larisa E. Seewordova ◽  
Yuriy R. Akhverdyan ◽  
Yuliya V. Polyakova
Keyword(s):  
Rheumatoid Arthritis ◽  
Proximal Femur ◽  
Type I Collagen ◽  
Osteoporotic Fractures ◽  
Type I ◽  
Mineral Density ◽  
Correlation Relationship ◽  
Fetuin A ◽  
The Mean ◽  
Total Alkaline

Aim. To study the level of fetuin-A (FA), bone mineral density (BMD) and certain markers of bone remodeling in patients with rheumatoid arthritis (RA). Materials and Methods. 110 Patients with RA and 30 conventionally healthy patients were examined. In both groups the levels of FA and BMD were determined. In the group of patients with RA the levels of C-telopeptide of type I collagen, N-terminal propeptide of procollagen I, 25-hydroxy-cholecalciferol, total alkaline phosphatase, total calcium of blood were determined. Results. The mean concentration of FA in blood serum of patients with RA was 765.67±120.66 µg/mL, which was below the respective parameter in donors – 812.95±76.21 µg/mL (p=0.0437). In the group of patients with RA with osteoporosis (n=52) the mean level of FA was 733.65±135.84 µg/mL, and in the group without osteoporosis – 794.37±97.7 µg/mL (p=0.0044). The level of FA was also reduced in patients with osteoporotic fractures (n=24) – 694.79±110.47 µg/mL against 785.45±116.43 µg/mL in patients without osteoporotic fractures (n=86; p=0.00091). A positive correlation relationship was found between the level of FA and BMD of L1-L4 (r=0.194; p=0.042), femoral neck (r=0.328; p<0.0001) and proximal femur (r=0.293; p=0.002), and the level of 25-hydroxy-cholecalciferol (r=0.259; p=0.006), and the negative correlation relationship – with C-telopeptide of type I collagen (r=-0.203; p=0.033). Conclusions. Patients with RA with reduced FA level were characterized by a higher detection rate of osteoporosis and of osteoporotic fractures, and by a lower BMD of L2-L4, femoral bone and proximal femur. Besides, a lower level of FA was associated with a lower level of 25-hydroxy-cholecalciferol and with a higher level of C-telopeptide of type I collagen which permits to suggest existence of osteoprotective function in this glycoprotein.

scholarly journals The role of calcification biomarkers in the formation of aortic stenosis

2021 ◽  
Vol 42 (Supplement_1) ◽  
Author(s):  
Y Sibagatullina ◽  
M Pugina ◽  
I Voronkina ◽  
E Zhiduleva ◽  
P Evstigneeva ◽  
...  
Keyword(s):  
Heart Disease ◽  
Aortic Valve ◽  
Aortic Stenosis ◽  
Blood Serum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Fetuin A ◽  
Positive Correlation ◽  
Funding Sources ◽  
Potential Methods

Abstract Background The main mechanism of aortic valve (AV) calcification is not yet known. One of the possibility pathway responsible for AV calcification (AVC) includes osteoprotegerin (OPG), receptor activator of nuclear factor kappa-B ligand (RANKL) – the parts of the RANKL-RANK-OPG system and fetuin-A: cysteine protease inhibitors. Aim Evaluation of OPG, sRANKL, fetuin-A in blood serum and tissues in patients with varying severity of aortic stenosis (AS) to establish potential methods of estimation AVC depending on the presence of congenital heart disease: bicuspid AV (BAV) or its absence. Materials and methods The study involved 285 patients with AS (59.06±6.95 years, m:f – 1.1:1): 163 with (BAV) and 122 with tricuspid AV (TAV). The control group 53 patients (48.31±9.04 years, m:f-1:1) without valvular pathology, connective tissue dysplasia and coronary heart disease. ECHO (Vivid 7, GE, USA) was performed in all patients. The expression of OPG, RANKL, fetuin-A in homogenates of aortic valve were determined by immunoblotting. Serum concentration of OPG, RANKL, fetuin were performed in all pts by enzyme-linked immunosorbent assay. Results In the control group the concentration of fetuin-A in the blood serum was significantly higher than in TAV and BAV subgroups (446.66 [293.63; 619.19] vs 319.9 [239.6; 400.2] vs 315.6 [245.6; 385.6] pmol/l, p=0.ehab724.1560001). In all groups of patients with AS an increased level of sRANKL in the blood serum was revealed compared to the control group (TAV=0.39 [0.25; 0.53] vs BAV=0.38 [0.21; 0.55] vs control group 0.31 [0.18; 0.44] pmol/l; p&lt;0.005). OPG level in serum was increased in patients with TAV: 8.1 [4.3; 11.9] pmol/l compared to BAV: 6.8 [3.9; 9.7] pmol/l, p=0.003, as well as the control group: 6.15 [3.41; 8.89] pmol/L, p=0.001. RANKL expression in AV tissue was significantly lower in patients in the control group: 2119,06 [1990.94; 2554.11] as compared with AS pts: 4273.03 [2887.620; 4956], p=0.001, and in subgroups with TAV: 4273.08 [2887.620; 5285], p=0.002 or BAV: 4272.99 [2884.430; 4847], p&lt;0.01. In addition, a positive correlation of moderate strength was found between the RANKL in the blood serum and the expression of RANKL in the AV tissue in patients with BAV (r=0.357, p=0.04). OPG expression in the AV tissue was higher in patients in the control group: 3949,953 [1931.88; 6447.67], while significant only in comparison with the BAV subgroup: 2599.28 [1066.38; 4132.18], p=0.02. A positive correlation of moderate strength was found between the OPG in the blood serum and the expression of OPG in the AV tissue in TAV subgroup (r=0.423, p=0.03). Conclusion Different pathogenic mechanisms of AV calcification are accompanied by an increase in various markers of the OPG / RANK / RANKL system: in patients with TAV calcification marker is OPG, in patients with BAV it is RANKL. FUNDunding Acknowledgement Type of funding sources: None.

Antibodies to enzymes of antioxidant system as a pathogenetic component of anemia in rheumatoid arthritis

2019 ◽  
pp. 96-100
Author(s):  
И.П. Гонтарь ◽  
О.И. Емельянова ◽  
О.А. Русанова ◽  
Л.А. Маслакова ◽  
И.А. Зборовская ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Magnetic Material ◽  
Superoxide Dismutase ◽  
Enzymatic Activity ◽  
Blood Serum ◽  
Glutathione Reductase ◽  
Protein Concentration ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Concentration ◽  
Glutaric Aldehyde

Цель исследования - изучение антителообразования к супероксиддисмутазе, глутатионредуктазе, каталазе у больных ревматоидным артритом и выяснение их роли в патогенезе сопутствующей анемии. Методика. Проведено обследование 104 больных ревматоидным артритом с различной степенью активности процесса, формой и характером течения. Диагноз ревматоидного артрита ставился на основании клинико-лабораторного и инструментального обследования больных согласно системе диагностических критериев Американской ревматологической ассоциации, предложенной в 2010 году (ACR / EULAR). Выраженность анемии оценивали по уровню гемоглобина и количеству эритроцитов. Определение уровня антител проводили методом непрямого иммуноферментного анализа [ELISA тест] с использованием иммобилизированных магнитосорбентов, представляющих собой полиакриламидные гранулы, содержащие магнитный материал и перечисленные ферменты в качестве антигена. В качестве антигенов использовались коммерческие отечественные препараты: супероксиддисмутаза из эритроцитов человека (активность 30 Ед/мг), в исследованиях использовали фермент в рабочем разведении по белку - 100 мкг/мл, глутатионредуктаза (активность 340 Ед/мг) - 200 мг/мл по белку, каталаза (активность 380 Ед/мг) - 1,4 мг/мл по белку. Учитывая достаточную относительную молекулярную массу глутатионредуктазы и каталазы, иммобилизацию проводили методом эмульсионной полимеризации в потоке газообразного азота с включением магнитного материала. В связи с небольшой относительной молекулярной массой эритроцитарной фиксации супероксиддисмутазы иммобилизацию фермента проводили путем пришивки его молекулы глютаровым альдегидом к инертной полиакриламидной грануле, содержащей магнитный материал. «Чистые» антитела к ферментам получали с помощью соответствующего антигенного иммуносорбента. Источником специфических иммуноглобулинов служили сыворотки больных ревматоидным артритом с заранее определенным высоким титром антител (экстинция>0,2). После инкубации антигенного иммуносорбента и растворимой формы фермента с полученными антителами был проведен анализ изменения активности энзима. Статистическую обработку результатов проводили с использованием программных пакетов Statistica 6.0, Excel 5.0, Statgraphics 3.0, SPSS 12.0. Результаты. Выявлена зависимость между уровнем антител к супероксиддисмутазе, глутатионредуктазе и каталазе и активностью, течением и формой болезни. Полученные результаты свидетельствуют о повышении антителогенеза к этим ферментам по мере активизации патологического процесса. Выявленное снижение активности энзимов, связанное с выработкой антител к ним, происходит, очевидно, вследствие блокирования специфическими иммуноглобулинами активного центра фермента, являющегося одновременно и антигенной детерминантой. Об участии антител к ферментам в патогенезе анемии у больных ревматоидным артритом свидетельствует обратная корреляция между содержанием антител и уровнем гемоглобина. У больных ревматоидным артритом с умеренно выраженной анемией содержание антител было значимо выше, чем у пациентов без анемии, но ниже, чем у пациентов с тяжелой формой анемии. Заключение. Антитела к супероксиддисмутазе, глутатионредуктазе и каталазе являются одним из патогенетических факторов развития анемии и могут служить критерием тяжести заболевания. The aim was to study formation of antibodies to superoxide dismutase (SOD), glutathione reductase (GR), and catalase (CAT) in patients with rheumatoid arthritis (RA) and to elucidate the role of the antibodies in development of anemia. Methods. The antibodies were determined by enzyme-linked immunosorbent assay (ELISA) using immobilized magnetic sorbents, polyacrylamide granules containing a magnetic substance and the enzymes (SOD, GR, CAT) as antigens. Antibody concentration was expressed in absorbance units. Rheumatoid arthritis was diagnosed based on clinical and instrumental evaluation of patients according to the criteria of the American College of Rheumatology (ACR/EULAR, 2010). Statistical analysis was performed using variation statistics, and results were expressed as mean±SEM. Central tendencies were compared using the Student’s test. Differences were considered statistically significant at p<0.05. ELISA was performed on blood serum from 104 rheumatoid arthritis patients with different disease activity. The following commercial reagents (Russia) were used as antigens: superoxide dismutase from human RBCs (30 U/mg), which was used in the study in a working protein dilution of 100 mcg/ml; glutathione reductase (340 U/mg) which was used in the study at a protein concentration of 200 mg/ml; and catalase (380 U/mg), which was used in the study at a protein concentration of 1.4 mg/ml. Since the relative molecular weights of glutathione reductase and catalase were sufficient, immobilization was performed by emulsion polymerization in a flow of gaseous nitrogen including a magnetic material. Due to a low relative molecular weight of SOD from RBCs this enzyme was immobilized by coupling to an inert polyacrylamide granule containing a magnetic material, using glutaric aldehyde. «Pure» antibodies to the enzymes were obtained using a respective antigen immunosorbent. Specific immunoglobulins were obtained from blood serum of rheumatoid arthritis patients with a known high antibody titer (extinction > 0.2). Enzymatic activity was analyzed following incubation of the antigen immunosorbent and soluble enzyme with the obtained antibodies. Results. Production of antibodies to the studied enzymes increased with increasing severity of the disease. The decrease in enzymatic activity associated with production of respective enzyme antibodies is apparently due to inhibition by specific immunoglobulins of the active center, which is also the antigenic determinant. Conclusion. Antibodies to SOD, GR, and CAT are pathogenetic factors in the development of anemia; they may serve as a criterion for disease severity.

Berberine Inhibits the gene Expression and Production of Proinflammatory Cytokines by Mononuclear cells in Rheumatoid Arthritis and Healthy Individuals

2020 ◽  
Vol 16 ◽  
Author(s):  
Niloofar Ghorbani ◽  
Maryam Sahebari ◽  
Mahmoud Mahmoudi ◽  
Maryam Rastin ◽  
Shahrzad Zamani ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Gene Expression ◽  
Mononuclear Cells ◽  
Inhibitory Effect ◽  
Healthy Individuals ◽  
High Concentration ◽  
Antiinflammatory Property ◽  
Tnf Α ◽  
Anti Inflammatory ◽  
Inflammatory Effect

Objective: Rheumatoid arthritis (RA) is the most prevalent autoimmune arthritis. Berberine is an alkaloid isolated from Berberis vulgaris and its anti-inflammatory effect has been identified. Method: Twenty newly diagnosed RA patients and 20 healthy controls participated. Peripheral mononuclear cells were prepared and stimulated with bacterial lipopolysachharide (LPS,1 µg/ml), exposed to different concentrations of berberine (10 and 50µM) and dexamethasone (10-7 M) as a reference. Toxicity of compounds was evaluated by WST-1 assay. Expression of TNF-α and IL-1β were determined by quantitative real-time PCR. Protein level of secreted TNF-α and IL1β were measured by using ELISA. Result: Berberine did not have any toxic effect on cells, whereas Lipopolysachharide (LPS) stimulation caused a noticeable rise in TNF-α and IL-1β production. Berberine markedly downregulated the expression of both TNF-α and IL1β and inhibits TNF-α and IL-1β secretion from LPS-stimulated PBMCs. Discussion: This study provided molecular basis for anti-inflammatory effect of berberine on human mononuclear cells through the suppression of TNF-a and IL-1secretion. Our findings highlighted the significant inhibitory effect of berberine on proinflammatory responses of mononuclear cells from rheumatoid arthritis individuals, which may be responsible for antiinflammatory property of Barberry. We observed that berberine at high concentration exhibited anti-inflammatory effect in PBMCs of both healthy and patient groups by suppression of TNF-a and IL-1cytokines at both mRNA and protein levels. Conclusions: Berberine may inhibit the gene expression and production of pro-inflammatory cytokines by mononuclear cells in rheumatoid arthritis and healthy individuals without affecting cells viability. Future studies with larger sample size is needed to prove the idea.

scholarly journals AB0327 SERUM IRISIN LEVELS IN HEALTHY WOMEN AND PATIENTS WITH RHEUMATOID ARTHRITIS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1462.1-1462
Author(s):  
A. Yury ◽  
В. Zavodovsky ◽  
P. J ◽  
S. L ◽  
E. Papichev ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Normal Level ◽  
Normal Values ◽  
Reference Interval ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Healthy Individuals ◽  
Average Value ◽  
Healthy Female ◽  
Group 2

Objectives:to study serum irisin levels in healthy females and patients with rheumatoid arthritis.Methods:We examined 110 patients with a reliable diagnosis of rheumatoid arthritis (RA). The age of the examined was from 18 to 69 years, all patients were female. The diagnosis of RA was established on the basis of the 2010 EULAR diagnostic criteria. The group of patients included patients with a diagnosis at least one month before the planned screening. As a control group, as well as to create a representation of the normal values irisina level in the blood serum of healthy persons were examined 60 healthy volunteers (all women). In both groups, the level of serum irisin was determined using the enzyme-linked immunosorbent assay by the commercial Irisin ELISA kit.Results:As a result of measurements in the group of healthy individuals, the average value with the standard deviation used to assess the reliability of the average values was 20.49±4.82 μg/ml (μ±σ). By calculation, a reference interval of 10.85-30.13 μg/ml was determined, defined as μ±2σ. In patients with RA, the level of serum irisin was 14.52±6.99 μg / ml (μ±σ), which is significantly lower than in healthy individuals (p<0.01). Then we divided all patients into 2 groups: group 1 with normal values (66 people), group 2 (44 patients) - with a reduced (less than 10.85 μg/ml) level of irisin. In both groups, the dynamics of the level of serum irisin was studied depending on the duration of the disease. Among patients with a disease RA duration of less than 4 years, 16 (24.24%) patients had a normal level of irisin, and 14 (31.82%) had a reduced level (less than 10.85 μg/ml). Among patients with a disease duration of more than 10 years, 36 (54.55%) patients had a normal level of serum irisin, and a low level was determined in 16 (36.36%) patients (χ2=3.568, p=0.168).Conclusion:According to the data obtained, the normal level of serum irisin in healthy female individuals was 10.85-30.13 μg/ml. In patients with RA, the average level of irisin is significantly lower, however, with an increase in the duration of the disease, it tends to normalize.Disclosure of Interests:None declared

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