Xanthine Oxidase Enzyme Activity Research Articles

Effect of konjac glucomannan on gut microbiota from hyperuricemia subjects in vitro: fermentation characteristics and inhibitory xanthine oxidase activity.

The disorder of uric acid metabolism is closely associated with gut microbiota and short-chain fatty acids (SCFAs) dysregulation, but the biological mechanism is unclear, limiting the development of uric acid-lowering active polysaccharides. Konjac glucomannan (KGM) could attenuate metabolic disturbance of uric acid and modulate the gut microbiota. However, the relationship between uric acid metabolism and gut microbiota is still unknown. In this study, The fecal samples were provided by healthy volunteers and hyperuricemia (HUA) patients. Fecal samples from healthy volunteers was regarded as the NOR group. Similarly, 10% HUA fecal suspension was named as the HUA group. Then, fecal supernatant was inoculated into a growth basal medium containing glucose or KGM, and healthy fecal samples were designated as the NOR-GLU and NOR-KGM groups, while HUA fecal samples were designated as the HUA-GLU and HUA-KGM groups. All samples were cultured in an anaerobic bag system. After fermentation for 24 h, the samples were collected for further analysis of composition of intestinal microbiota, SCFAs concentration and XOD enzyme activity. The results showed that KGM could be utilized and degraded by the gut microbiota from HUA subjects, and it could modulate the composition and structure of their HUA gut microbiota to more closely resemble that of a healthy group. In addition, KGM showed a superior modulated effect on HUA gut microbiota by increasing Megasphaera, Faecalibacterium, Lachnoclostridium, Lachnospiraceae, Anaerostipes, and Ruminococcus levels and decreasing Butyricicoccus, Eisenbergiella, and Enterococcus levels. Furthermore, the fermentation solution of KGM showed an inhibitory effect on xanthine oxidase (XOD) enzyme activity, which might be due to metabolites such as SCFAs. In conclusion, the effect of KGM on hyperuricemia subjects was investigated based on the gut microbiota in vitro. In the present study. It was found that KGM could be metabolized into SCFAs by HUA gut microbiota. Furthermore, KGM could modulate the structure of HUA gut microbiota. At the genus level, KGM could decrease the relative abundances of Butyricicoccus, Eisenbergiella, and Enterococcus, while Lachnoclostridium and Lachnospiraceae in HUA gut microbiota were significantly increased by the addition of KGM. The metabolites of gut microbiota, such as SCFAs, might be responsible for the inhibition of XOD activity. Thus, KGM exhibited a superior probiotic function on the HUA gut microbiota, which is expected as a promising candidate for remodeling the HUA gut microbiota.

ANTIHIPERURESEMIA FLAVONOID DARI EKSTRAK ETANOL PELEPAH DAUN NIPAH (Nypa fruticans.Wurmb)

Hyperuresimia or gout in advanced stages causes joint damage (gout), kidney stones and kidney damage. Flavonoids as antioxidants can inhibit the activity of the enzyme xanthine oxidase which causes uric acid not to form. Nypa fruticans or known as nipah is traditionally used in the treatment of asthma, leprosy, tuberculosis, sore throat, liver disease, snake bites, as a pain reliever, as a sedative and carminative. The content of flavonoid compounds in nipah plants includes chlorogenic acid, protocatechuic acid, kaempferol, gallic acid, hydroxybenzoic acid, rutin, quercetin, cinnamic acid, and anthocyanins. The purpose of this study was to isolate flavonoid compounds that have potential as antihyperuresemia from nipah fronds. The research process began with the extraction of nipah leaves using 96% ethanol by maceration, fractionation using liquid-liquid partition method, isolation using centrifugal chromatography (chromatotron) method and structure elucidation using 1HNMR, 13CNMR, FTIR and GC-MS. Antihyperuresemia testing was carried out in-vitro by inhibiting xanthine oxidase enzyme activity. From the results of the study, two flavonoid compounds were successfully isolated including (-) epicatechin, and chamferol. The results of measuring the inhibitory activity of xanthine oxidase enzyme obtained IC50 values of (-) epicatechin and chamferol of 3.543 ppm and 10.383 ppm, respectively.

P011 Altered Expression and Enzymatic Activity of Xanthine Oxidase in Inflammatory Bowel Disease

Abstract Background Xanthine oxidase (XO) is involved in the last steps of purine metabolism; indeed, it catalyzes the oxidation of hypoxanthine to xanthine and xanthine to uric acid (UA) with the production of superoxide anion. The accumulation of UA has been shown to initiate the inflammatory process through NLRP3 inflammasome and the production of reactive oxygen species (ROS) contributes to inflammation-related tissue damage. In murine models of colitis, XO inhibitors, allopurinol or febuxostat, reduced the expression of proinflammatory cytokines supporting the involvement of XO in the intestinal inflammatory process. This study aims to conduct a thorough investigation of the role XO plays in IBD pathogenesis and to explore the potential of XO inhibition as a therapeutic strategy. Methods Biopsies from the colon of patients with ulcerative colitis (UC) and Crohn’s disease (CD) were collected at University Hospital of Cagliari. Disease activity was assessed using Mayo and SES-CD scores. XO expression levels were evaluated using Real-Time PCR, validated by GEO database (GSE11223, GSE117993). XO enzymatic activity was quantified spectrophotometrically in protein extracts from the biopsies. XO and NLRP3 inflammasome activation pathway, including NLRP3, ASC, and CASPASE1, was investigated using immunohistochemistry (IHC). In vitro, Caco2, HT29, SW480 cells were treated with xanthine, XO inhibitors (allopurinol, febuxostat) to assess XO inhibition on NLRP3 activation, analyzed by immunocytochemistry (ICC). Results Biopsies from the sigmoid colon of 15 HC, 20 UC, and 15 CD patients, as well as from the ileum of 10 HC and 10 CD, were analyzed. Increased XO expression was found in UC and CD sigmoid mucosa (Relative Quantification [RQ]: 1.93 vs. 0.99, p=0.004), and CD ileum (RQ: 2.72 vs. 1.26, p=0.004), confirmed by GEO data. XO activity was higher in UC and CD sigmoid (3.86 nmol UA/min/mg in UC, 4.82 in CD vs. 1.03 in HC, p<0.001) and CD ileum (8.41 vs. 1.83 in HC, p<0.001). IHC confirmed that XO and the NLRP3 inflammasome pathway are more highly expressed in IBD patients than HC, predominantly within epithelial cells. In vitro studies on Caco2, HT29, and SW480 cells demonstrated that suppressing XO activity with XO inhibitors leads to the inhibition of the NLRP3 pathway. Conclusion XO expression was higher in IBD pts than in HC individuals. Alongside this increase in XO activity, there was a concurrent hyperactivation of the NLRP3 inflammasome. The inhibition of XO activity in cell lines resulted in reduced activation of the NLRP3 inflammasome pathway. These findings support the hypothesis that XO plays a critical role in the pathogenesis of IBD and highlight its potential as a therapeutic target.

Content of Phenols, Flavonoids and Antihyperuricemic Activity of Leaves And Rhizomes of Smilax Purhampuy Ruiz

Introduction: Smilax purhampuy is traditionally used for the treatment of chronic gastritis, cystitis, arthritis and inflammation of the prostate; however, it lacks scientific studies that support its traditional use. Objective: To estimate the chemical composition and antihyperuricemic activity of hydroalcoholic extracts from leaves and rhizomes of S. purhampuy. Method: the dry material of leaves and rhizomes of the species was used for the elaboration of the extracts by maceration with 80 % hydroalcoholic solvent. Total phenols were quantified by Folin-Ciocalteu and total flavonoids by the aluminum chloride colorimetric method. Antihyperuricemic activity was evaluated in rats using the potassium oxonate induction model; the extracts were tested at doses of 125, 250 and 400 mg/kg and allopurinol 10 mg/kg was used as a positive control. The uric acid content in blood serum was quantified and the enzymatic activity of xanthine oxidase was determined. Results: in the phytochemical determinations, significant differences were observed in the content of each evaluated metabolite, fundamentally associated with the characteristics of each plant organ. The highest content of phenols (4.08 mg EAG/g) and flavonoids (1.01 mg EQ/g) was found in the leaf extract. The extracts of leaves and rhizomes showed a significant decrease in the levels of uric acid in blood serum (156.31 µmol/L y 167.58 µmol/L, respectively) and in the activity of the enzyme xanthine oxidase at all doses tested, being more marked in the extract of leaves at 400 mg/kg. Conclusions: the results provide the first findings of the antihyperuricemic potential of the species and make an important contribution to the knowledge of its chemical composition

Inhibiting Hepatocyte Uric Acid Synthesis and Reabsorption Ameliorates Acetaminophen-Induced Acute Liver Injury in Mice

Acetaminophen (APAP) overdose is the most common cause of drug-induced liver injury worldwide. Uric acid (UA) is involved in sterile inflammation in many organs, but its role in APAP-induced liver injury remains elusive. We quantified the concentration of UA in the serum and liver tissues of APAP-overdosed mice and explored the changes in proteins involved in UA synthesis, absorption, and degeneration on APAP stimulation. We also examined the effects of inhibiting hepatocyte UA synthesis or reabsorption on APAP-induced liver injury in mice. Furthermore, we explored the process of UA clearance by peripheral macrophages. APAP overdose significantly increased intrahepatic UA contents, which occurred earlier than apparent hepatocyte injury in APAP-overdosed mice. APAP overdose induced significant DNA leakage and may thereby increase the substrate of UA synthesis. APAP overdose also significantly increased the enzymatic activity of xanthine oxidase and urate oxidase and decreased the expression of the UA reabsorption transporter GLUT9 in hepatocytes. Inhibiting hepatocyte UA synthesis by febuxostat or reabsorption by hepatic-specific knockout of GLUT9 alleviated APAP-induced liver injury. Further experiments showed that monosodium urate but not soluble UA may be a major form of UA mediating hepatocyte injury. Additionally, monosodium urate further recruited circulating macrophages into the liver and then aggravated inflammation by increasing the levels of inflammatory factors and reactive oxygen species. Deletion of macrophages significantly ameliorated APAP-induced liver injury in mice. APAP overdose induces excessive UA production and leads to local high concentrations in the liver, which further injures cells and induces liver inflammation. Inhibiting the production of UA may be a potential therapeutic option for treating APAP-induced liver injury.

ANTIHYPERURESEMIA ACTIVITY OF ETHANOL EXTRACTS AND FRACTIONS OF NIPAH LEAF FRONDS (NYPA FRUTICANS. WURMB)

Hyperuresemia is a metabolite syndrome characterised by high levels of uric acid in the blood. Hyperuresmia in advanced stages can cause gout or pyrexia (a disease that affects the joints and causes joint damage), kidney stones, kidney damage, and hypertension. The nipah plant (Nypa fruticans. Wurmb) is traditionally used in the treatment of stomach pain, diabetes, fever, toothache, uric acid reduction and headache medicine. In general, the content of secondary metabolite compounds from nipah plants includes flavonoids, phenols, tannins, terpenois, steroids, alkaloids and saponins. The purpose of this study was to determine the xanthine oxidase inhibitory activity of ethanol extracts and fractions of nipah leaves (Nypa fruticans. Wurmb). Extraction process using maceration method with 96% ethanol solvent, fractionation using liquid-liquid partition method with separating funnel. . Testing the inhibition of xanthine oxidase enzyme activity in vitro using UV/VIS spectrophotometer. The results showed that ethanol extract, water fraction, ethyl acetate fraction and n-hexane fraction showed inhibitory activity of xanthine oxidase enzyme with IC50 values of 47.558 ± 2.862; 18.755 ± 3.821; 3.543 ± 1.440 and 110.213 ± 1.577 ppm, respectively. From the results of the study it can be concluded that ethanol extracts and fractions of nipah leaf fronds (Nypa fruticans.Wurmb) can inhibit the activity of xanthine oxidase enzyme with ethyl acetate fraction giving the highest inhibitory activity with IC50 value of 3,543±1,440 ppm.

Effect of kaffir lime (Cytrus hystrix) leaf extract on xanthine oxidase inhibition

The xanthine oxidase enzyme is an enzyme that catalyzes purine metabolic reactions, the oxidation of hypoxanthine to uric acid through xanthine compounds. High uric acid causes hyperuricemia. One treatment is to inhibit the xanthine oxidase enzyme. It can be conducted with a natural remedy, for example kaffir lime leaf extract. Research showed that kaffir lime leaves contain flavonoids that can inhibit the activity of the xanthine oxidase. This research aimed to examine kaffir lime leaf extract as an inhibitor of xanthine oxidase enzyme activity. Isolation of kaffir lime leaves was conducted by various maceration times of 24, 48, and 72 hours. The method of determining the inhibition value (IC50) of kaffir lime leaf extract to the xanthine oxidase enzyme was UV-Vis spectrophotometer at λ 291,5 nm. Maceration results of kaffir lime leaf extract with phytochemical tests for 48 hours showed that there were phenolic, flavonoids, steroids, alkaloids, tannins, and triterpenoids compounds. FT-IR test showed that there were C=O, C=C aromatic, O-H, and CH-aliphatic groups, which indicated the structure of the flavonoids. Maceration of kaffir lime leaf extract for 48 hours was a weak inhibitor with an IC50 value of 285,82 ppm.

Hydro-alcoholic extract of Ochradenus baccatus exhibits anti-oxidative and anti-inflammatory properties and inhibits enzyme of uric acid metabolism: Implications for bioactive molecules

Traditional medicines have been an accessible, affordable, and culturally acceptable model of healthcare that is trusted acrossthe globe in terms of disease management, symptoms relief and patient satisfaction. The natural products derived from plant sources arethe fundamental ingredients of traditional and folk medicine and serve as lead compounds or pharmacophore for most effective drugs inmodern pharmacology for treatment of infectious and chronic diseases. Ochradenus baccatus, which belongs to family Resedaceae, is aperennial shrub, widely distributed in the arid regions of Arabian Peninsula and finds mention in the traditional medicine for reproductivehealth, inflammation and infection. Results: As a proof-of-concept, this study has investigated the various bioactive properties of the plantvis-à-vis anti-oxidative, anti-inflammatory and inhibition of xanthine oxidase using standard in vitro experimental models. The bioactivemolecules found in the extract included flavonoids, phenolics, tannins, saponins and alkaloids in varying quantities, which are known tobe associated with a multitude of pharmacological properties. Our results show that the extract caused a progressive inhibition of DPPHradical and neutralizes superoxide anions in a dose-dependent manner. The extract demonstrated its redox-active potential by means of itsability to reduce Cu (II) to Cu (I) that is detected using bathocuproine, a Cu (I)-specific sequestering agent. Progressive doses of the extractsinhibited the denaturation of protein (a factor associated with inflammation), with maximum inhibition of 70.6% observed at concentrationof 600 μg/ml, after which the effect plateaued for further doses. Moreover, the extract displayed a progressive inhibitory property againstthe enzymatic activity of xanthine oxidase with a maximum inhibition of 66% observed at a dose of 300 μg/ml. Conclusion: Inhibition ofxanthine oxidase reduces the hyperuricemia in patients with gout and nephrolithiasis as it alleviates the tendency of salt accumulation inthe joints and kidney. The interest in the natural products pharmacology is escalating and exploring novel bioactive properties in plantsof interest is a significant stride from the traditional knowledge to evidence-based practice

L-Citrulline Supplementation Reduces Blood Pressure and Myocardial Infarct Size under Chronic Intermittent Hypoxia, a Major Feature of Sleep Apnea Syndrome.

Intermittent hypoxia (IH) is a landmark of obstructive sleep apnea (OSA) at the core of the cardiovascular consequences of OSA. IH triggers oxidative stress, a major underlying mechanism for elevated blood pressure (BP) and increased infarct size. L-citrulline is an amino acid that has been demonstrated to be protective of the cardiovascular system and exert pleiotropic effects. Therefore, we tested the impact of citrulline supplementation on IH-induced increase in BP and infarct size. Four groups of rats exposed to normoxia (N) or IH [14 days (d), 8 h/day, 30 s-O2 21%/30 s-O2 5%] and were supplemented or not with citrulline (1 g·kg-1·d-1). After 14 d, BP was measured, and hearts were submitted to global ischemia-reperfusion to measure infarct size. Histological and biochemical analyses were conducted on hearts and aorta to assess oxidative stress. Citrulline significantly reduced BP (-9.92%) and infarct size (-18.22%) under IH only. In the aorta, citrulline supplementation significantly decreased superoxide anion and nitrotyrosine levels under IH and abolished the IH-induced decrease in nitrite. Citrulline supplementation significantly decreased myocardial superoxide anion levels and xanthine oxidase enzyme activity under IH. Citrulline shows a cardioprotective capacity by limiting IH-induced pro-oxidant activity. Our results suggest that citrulline might represent a new pharmacological strategy in OSA patients with high cardiovascular risk.

Drug-Drug Interaction of Aldehyde Oxidase Inhibitor and Xanthine Oxidase Inhibitor with Favipiravir

Aim: Favipiravir is an effective antiviral used in the treatment of COVID-19. It is metabolized by aldehyde oxidase (AO) and xanthine oxidase (XO). This study investigated drug-drug interactions between favipiravir with both AO substrate and XO enzyme inhibitor, allopurinol, and an XO inhibitor, verapamil. 
 Material and Methods: 25 Sprague-Dawley female rats, 250-300 g, were divided into five equal groups. Blood samples were taken from the jugular vein at the end of 0, 15, 30, and 45 minutes, and at the end of the 1st, 2nd, 4th, 6th, and 8th hours after the drugs were administered. The drug-blood concentration was determined in the HPLC-UV device using plasma. The ELISA method measured AO and XO enzyme activities in rat liver tissue.
 Results: Allopurinol prolonged the time taken for favipiravir to reach Cmax (Tmax), decreased maximum serum concentration (Cmax), elimination half-life (T1/2), area under the curve (AUC), and mean residence time (MRT). Allopurinol significantly reduced clearance per unit time (Cl/f) when co-administered with favipiravir. Verapamil accelerated the elimination of favipiravir, significantly reducing T1/2, MRT, and AUC. On the other hand, Favipiravir decreased the absorption of verapamil and slowed its elimination. Cmax, AUC, and Cl values of verapamil decreased. In addition, T1/2, MRT, and volume of distribution (Vd) increased. 
 Conclusion: In conclusion, the concomitant use of favipiravir with other drugs that affect AO and/or XO enzyme activities may cause changes in the pharmacokinetic profiles of drugs and the levels of enzymes that metabolize drugs.

Characterization of xanthine oxidase inhibitory activities of phenols from pickled radish with molecular simulation

Pickled radish is a general source of natural bioactive compounds that include phenols. Here, we used molecular docking, fluorescence quenching, circular dichroism spectroscopy and molecular dynamics simulations to identify potential inhibitors against xanthine oxidase from a library of pickled radish compounds. The most effective compounds were selected for validation through in vitro experiments including enzyme activity inhibition tests, and cell-based assays. Molecular docking results revealed that 2,6-Dihydroxyacetophenone, 4-Hydroxyphenethyl alcohol, and 4-Hydroxybenzaldehyde exhibited significant effects on xanthine oxidase inhibition. Three phenols have varying degrees of inhibition on xanthine oxidase, which is driven by hydrophobic interactions and hydrogen bonds and affects the secondary structure and hydrophobic homeostasis of xanthine oxidase. The stability of xanthine oxidase inhibition by three phenols was analyzed by molecular dynamics simulation. Finally, cellular experiments confirmed that three phenols reduced uric acid levels by inhibiting the xanthine oxidase enzyme activity of BRL 3A cells.

Antioxidant and Xanthine Oxidase Inhibitory Potential of Aqueous Extract of Ferula Asafoetida

Background: Ferula asafoetida is a dried latex that is exuded from rhizome or taproot species. Organosulfides are primarily responsible for flavour and odour of asafoetida.Ferula asafoetida is a natural medicine good for asthma and bronchitis. is also used to relieve stomach gas, digestive issues. It is usually added while cooking to hormonise the sweet, sour, salty, spicy taste of the food. Increased activity of xanthine oxidase is involved in the medical condition known as gout, which is characterized by hyperuricemia that leads to deposition of uric acid in the joints resulting in painful inflammation.
 Aim: To analyse the anti-oxidant and xanthine oxidase inhibitory potential of aqueous extract of Ferula asafoetida.
 Materials and Methods: Preparation of the aqueous seed extract of Ferula asafoetida done by hot percolation method. Phytochemical screening, in vitro antioxidant activity and xanthine oxidase inhibitory potential was done by standard procedures. The data were analyzed statistically by a one - way analysis of variance (ANOVA) followed by Duncan’s multiple range test was used to see the statistical significance among the groups. The results with the p<0.05 level were considered to be statistically significant.
 Results: The phytochemical screening revealed that the extract is rich in phytoconstituents. DPPH radical scavenging activity established the potent in vitro antioxidant activity (p<0.05) of Ferula asafoetida extract. The extract was also efficient in inhibiting the activity of xanthine oxidase enzyme (p<0.05) in a concentration dependent manner.
 Discussion: The extract has potent antioxidant and xanthine oxidase inhibitory potential, although the activities are less compared to the standard drug.
 Conclusion: The Ferula asafoetida extract can be used to treat gout and to combat various other disorders associated with xanthine oxidase activity.

Vaccinium virgatum Aiton Leaves Extract Suppressed Lipid Accumulation and Uric Acid Production in 3T3-L1 Adipocytes.

Blueberry (Vaccinium virgatum Aiton; Kinisato 35 Gou) leaves have recently attracted increasing attention as a useful material for the prevention of lifestyle diseases. Here, we examined the effects of the hot water extract of blueberry leaves (BLEx) on lipogenesis and uric acid production in 3T3-L1 adipocytes. The results showed that BLEx suppressed lipid accumulation and the mRNA expression of differentiation markers in 3T3-L1 adipocytes. A fractionation study showed that the highly polymerized proanthocyanidin-rich fraction was responsible for this effect. Upon maturation to adipocytes, 3T3-L1 cells produced uric acid and tumor necrosis factor-α, and hypoxia stimulated the production of uric acid and xanthine oxidoreductase activity. BLEx suppressed the production of uric acid under these conditions. Although BLEx inhibited the enzymatic activity of xanthine oxidase, this activity was observed in several fractions containing catechin, epicatechin, chlorogenic acid, rutin, and low molecular weight proanthocyanidins. Taken together, these results indicate that BLEx contains various compounds with the ability to suppress lipid accumulation and uric acid production in adipocytes.

Therapeutic antigout and antioxidant activity of Piper betle L. in gout-induced broilers

ABSTRACT 1. The following trial investigated the antigout activity and probable mechanism of Piper betle L. herb in gout-induced broiler chickens. The antioxidant and production performance modulating activity of P. betle L. was compared against the standard antigout drug Allopurinol. 2. One hundred and sixty, one-day-old female chicks were randomly divided into five treatment groups (control, gout challenged, Allopurinol, P. betle 20 g/kg and P. betle 25 g/kg of feed) with eight birds per group (four replicates) and fed over six weeks. Gout was induced using sodium bicarbonate in water (20 g/l). The clinical signs of gout and production performance were recorded and gross and histopathology was conducted on dead birds. Serum uric acid and creatinine were estimated (on d 10, 14, 17, 19, 21 and 42) and antioxidant and xanthine oxidase enzyme activities were measured from blood samples. 3. Uric acid progressively reduced after treatment with P. betle 20 g/kg from d 17 (19.4 ± 0.62 mg/dl) to d 21 (9.81 ± 0.3 mg/dl) and xanthine oxidase activity was likewise suppressed (7.80 ± 0.04 U/mg protein), in a similar manner to Allopurinol (7.75 ± 0.05 U/mg protein), which authenticated the mechanism of antigout activity. Better feed conversion ratios (1.83 ± 0.001) and the restoration of the antioxidants superoxide dismutase, catalase and glutathione to normal levels were observed from birds fed P. betle 20 g/kg than with Allopurinol. 4. The data showed that P. betle can be an effective treatment for gout in broiler chicken, as an alternative to Allopurinol.

Features of disorders of uric acid metabolism in male patients with hypertension and atrial fibrillation

Abstract Background In recent decades, a significant number of epidemiological studies showed the association between asymptomatic hyperuricemia and development of arterial hypertension (HTN) and atrial fibrillation (AF). The key enzyme participating in disintegration purines to uric acid is xanthine oxidase (XO). The accumulated data suggest the existence of a direct pathophysiological role of increased level of serum uric acid (SUA) and XO activity with a risk of HTN and AF. Purpose To evaluate the features of disorders of UA metabolism in patients with HTN and AF. Materials and methods The study included male patients 27 to 65 years, with a body mass index (BMI) less than 30 kg/m2 and more than 18.5 kg/m2. All patients were investigated of the level of a SUA, hypoxanthine/xanthine, inosine/adenosine, XO enzyme activity. Depending on the presence of AF, all patients were divided into 2 groups: group 1 (n=16) patients with HTN, group 2 (n=16) patients with HTN and AF. Results Table 1 shows the indicators of purine metabolism in the compared groups. The analysis of the results demonstrates that in group 1 the smallest level of a SUA in comparison with group 2 (0.35 [0.31; 0.41] vs 0.41 [0.39; 0.40](μmol/l), p=0.003), hypoxanthine 3.02 [1.80; 5.63] vs 7.35 [4.56; 8.25](μmol/l), p=0.008; inosine 1.59 [1.25; 4.48] vs 7.41 [1.49; 17.24](μmol/l), p=0.002. An increase level of a SUA is noted in 31.2% of patients from group 1 (5 patients out of 16) and 68.7% of patients from group 2 (11 patients out of 16). As a result of Spearman's correlation analysis in the group of patients with HTN and AF, a direct correlation was found between the experience of AF and BMI (r=0.681), SBP (r=0.487); level of hypoxanthine and inosine (r=0.808) and the inverse correlation between the level of xanthine and inosine (r=−0.547), as well as the association between the level of XO and SUA (r=0.796), XO and BMI (r=0.812). In the group of patients with HTN was determined association between the level of hypoxanthine and inosine (r=0.604), xanthine and adenosine (r=0.539), inosine and adenosine (r=0.657), a direct association between the level of XO and adenosine (r=0.642), inosine (r=0.502), BMI (r=0.791). Conclusions As a result of the study of patients with isolated HTN in combination with AF at the age of 27 to 65 years and a BMI of less than 30 kg/m2 and more than 18.5 kg/m2, an increase level of SUA and intermediate metabolites of UA synthesis such as inosine and hypoxanthine is observed compared with group with isolated HTN. An increase in level of XO is associated with an increase level of SUA and BMI. In the group of patients with HTN, an increase in level of inosine is associated with an increase level of hypoxanthine, but an increased level of inosine is associated with a decrease in level of xanthine. The XO level in this group is associated with an increase in level of adenosine and inosine, BMI. Funding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): Grodno State Medical University, Grodno, Belarus Table 1. Comparative characteristics of indicators in groups 1 and 2

ANTIHYPERURICEMIA ACTIVITY OF ETHANOL EXTRACT AND FRACTIONS OF AZADIRACHTA INDICA LEAF IN VIVO AND MECHANISM OF ACTION OF ACTIVE FRACTIONS IN VITRO

Azadirachta indica , A.,Juss is a medicinal plant that is used traditionally for some disease, especially its leaves to treat a rheumatic diseases and lower blood uric acid levels. This study was carried out to examine antihyperuricemia activity of ethanol extract, water fraction, ethyl acetat fraction and n-hexane fraction of the A. Indica leaves in male mice of Swiss-Webster strain. Extract and fraction doses used were 250 and 500 mg/kg of body weight and the doses of allopurinol as a standard drug was 13 mg/kg of body weight. The tests were conducted on mice suffering from hyperuricemia induced by potassium oxonat at adose of 300 mg/kg of body weight intraperitoneally and chicken liver juice orally. Measurement of blood uric acid levels were performed using an Easy Touch® every hour for 4 hours after being given test preparations. The results showed that the ethanol extract and the fractions lowered blood uric acid levels in the same way as allopurinol did. The n-hexane fractions at the all doses showed the highest activity, followed by ethanol fraction, and water fraction at the dose of 250 mg/kg at the 4th hour. These results illustrated that the A. Indica leaves might be potential to be used as antihyperuricemia. The active compounds which possibly reduce blood uric acid levels in mice are believed to be flavonoid or polyphenolic compounds because they are reported to be able to inhibit the action of the xanthine oxidase enzyme that converts purines into uric acid. Then, the most active fraction, n-hexane fraction, was tested for its inhibitory activity on xanthine oxidase enzyme to determine its mechanism of action. The results showed that the n-hexane fraction, like allopurinol, inhibited uric acid biosynthesis through inhibiting the activity of xanthine oxidase enzyme with the IC 50 value of 132 μg/mL lower than that of allopurinol IC 50 58.35 μg/mL. Key words: A. indica leaves, antihyperuricemia, chicken liver juice, potassium oxonat, xanthine oxidase.

Assessment of Antigout, Antioxidant and Production Performance Augmenting Activity of Piper betle L. in Gout Induced Broiler Chicken

Background: The gout is commonly reported in birds and causes severe economic losses to the farmers. Hence to prevent the gout occurrence P.betle L. herb was selected and its antigout potential was studied in comparison with Allopurinol.Methods: The biological experiment was conducted with 120 broiler chicks and was divided into 5 treatment groups of 8 birds each with triplicate study for six weeks. The groups were control (T1), gout control (T2), Allopurinol (T3), P.betle L. - 10 g/kg (T4) and P.betle L. - 12.5 g/kg of feed (T5). The gout was induced using toxic dose of sodium bicarbonate in water (20 g/litre). The clinical signs and production performance were recorded and gross and histopathology studies were conducted. Serum biochemical parameters viz., uric acid and creatinine were periodically estimated (day 10, 15, 18 and 42 of trial) and anti-oxidant and xanthine oxidase enzyme activity were measured.Result: The production performances and the antioxidant activity were improved in P.betle L. groups and even better than Allopurinol. Also progressive reduction of uric acid and equivalent suppression of xanthine oxidase activity as that of Allopurinol were observed. Hence P.betle L. herb can be used effectively in the prevention of gout in birds.

Anti-inflammatory effects of functional beverage from a mixture of moringa leaves, pandanus leaves, and red ginger in mice induced with monosodium urate crystal

Production of uric acid that exceeds normal limits in the blood (hyperuricemia) can cause the formation of urate crystals and inflammatory reactions. In various studies, moringa leaves, pandanus leaves, and red ginger are known to contain phytochemical components (phenols and flavonoids) that are useful as antioxidants, anti-inflammatory, inhibiting xanthine oxidase enzyme activity, overcoming rheumatism, and antidiabetic. Functional drinks from the mixture of moringa leaves, pandanus leaves, and red ginger were optimised using the response surface method (RSM) in previous studies. The effect of this functional beverage product was tested for its effect as an anti-inflammatory in vivo. The testing process of the anti-inflammatory effect was using intradermally monosodium urate (MSU)-induced mice. Mice were measured for changes in edema, and after two days of treatment, the spleen was taken for flow cytometry testing of inflammatory cytokine expression of CD11b+ TNFα+, CD11b+ IL6+, CD11b+ IL10+, and blood was taken to test the expression of cytokine CD11b+ TNFα+. The results showed functional beverage products could potentially be used as an alternative anti-inflammatory agent in gout because it significantly inhibited edema, significantly inhibited the expression of proinflammatory cytokines CD11b+ TNFα+ and CD11b+ IL6+ and increased the expression of anti-inflammatory cytokines CD11b+ IL10+, where the anti-inflammatory effect was not significantly different from the control of indomethacin drugs.

Xanthine oxidase enzyme activity in keratoconic corneal epithelium.

To assess the activity of xanthine oxidase (XO) enzyme in keratoconic corneal epithelium and to evaluate its relationship with the keratoconus (KC) severity. This prospective and randomized study included 66 eyes of 54 KC patients who received corneal collagen cross-linking treatment and 43 eyes of 32 patients who underwent photorefractive keratectomy due to their refractive error. During surgical procedures, the corneal epithelium was mechanically scraped and gathered to analyze the XO enzyme activity spectrophotometrically. The KC group was subdivided into three groups (stages 1, 2, and 3) according to the Amsler-Krumeich classification. The results were compared between the KC and the control group and in between KC subgroups. No significant differences in age and gender were found between the KC and control groups (p = 0.064 and p = 0.296, respectively). The mean XO activity levels of the KC and control groups were 173.57 ± 87.61 and 223.70 ± 99.52mIU/mg, respectively (p < 0.001). In KC group, 33 eyes were at stage 1, 19 were at stage 2, and 14 were at stage 3. No significant difference was observed between KC subgroups regarding XO activity levels (p = 0.681). In this study, our findings revealed that ultraviolet-related pro-oxidant XO enzyme may have a role in the etiopathogenesis of KC. Further studies are needed to support our result. When we started this study in 2018, we did not have a "Clinical Trials Registration." However, we have ethics committee approval (date: 21. 02. 2018/No: 22).

Вплив лазерного опромінення на ксантиноксидазну активність та генерацію супероксидного радикала в цитозольній фракції печінки щурів-пухлиноносіїв

The effect of tumor growth in the body and laser irradiation on the enzymatic activity of xanthine oxidase, in particular its D- and O-forms, and also the rate of generation of the superoxide radical (O2–) and the level of protein sulfhydryl groups in the liver rat cytosolic fraction has been investigated. It has been found that in the cytosolic fraction of rats with transplanted Guerin’s carcinoma decreases the enzymatic activity of the D-form of xanthine oxidase with a simultaneous increase in its O-form during the period of intensive (14 days, which corresponds to the logarithmic phase of on cogenesis) and the period of final tumor growth (21 days, which corresponds to the stationary phase of oncogenesis). The increase in the enzymatic activity of the O-form of xanthine oxidase was accompanied by an increase the rate of superoxide radical generation and a decrease in the level of protein SH-groups in the liver cytosolic fraction of tumor-bearing rats. Daily directed action of laser irradiation on the area of growth of Guerin’s carcinoma leads to less destructive changes in the liver. Thus, there is an increase in the enzymatic activity of the D-form of xanthine oxidase, a decrease the rate of superoxide radical formation and an increase the content of protein SH-groups in the cytosolic fraction of the liver of experimental animals compared with non-irradiated tumor-bearing rats.