The Effects of Prolonged Treatment with Cemtirestat on Bone Parameters Reflecting Bone Quality in Non-Diabetic and Streptozotocin-Induced Diabetic Rats.

Cemtirestat, a bifunctional drug acting as an aldose reductase inhibitor with antioxidant ability, is considered a promising candidate for the treatment of diabetic neuropathy. Our study firstly examined the effects of prolonged cemtirestat treatment on bone parameters reflecting bone quality in non-diabetic rats and rats with streptozotocin (STZ)-induced diabetes. Experimental animals were assigned to four groups: non-diabetic rats, non-diabetic rats treated with cemtirestat, diabetic rats, and diabetic rats treated with cemtirestat. Higher levels of plasma glucose, triglycerides, cholesterol, glycated hemoglobin, magnesium, reduced femoral weight and length, bone mineral density and content, parameters characterizing trabecular bone mass and microarchitecture, cortical microarchitecture and geometry, and bone mechanical properties were determined in STZ-induced diabetic versus non-diabetic rats. Treatment with cemtirestat did not affect all aforementioned parameters in non-diabetic animals, suggesting that this drug is safe. In diabetic rats, cemtirestat supplementation reduced plasma triglyceride levels, increased the Haversian canal area and slightly, but insignificantly, improved bone mineral content. Nevertheless, the insufficient effect of cemtirestat treatment on diabetic bone disease does not support its use in the therapy of this complication of type 1 diabetes mellitus.

High-fructose intake-induced dyslipidemia and oxidative stress accompanied by hippocampal dysfunctions in hypertensive but not hypertriacylglycerolemic rats.

A high-fructose intake is metabolically analogous to a high-fat diet. The impact of highfructose intake was investigated in spontaneously hypertensive (SHR) and hypertriacylglycerolemic (HTG) rats to find out the impact of which risk factor of metabolic syndrome - hypertension or hypertriacylglycerolemia - will cause more complications. Rats were fed a standard or a fructose diet (F60) with 60% of added fructose for 5 weeks. The F60 diet increased the total serum cholesterol content of both HTG-F60 and SHR-F60 rats. Further, in SHR-F60 it increased serum triacylglycerols, TBARS in the liver, a specific activity of NAGA in the kidney, aggravated glucose tolerance, deteriorated synaptic plasticity, and reduced somatic and dendritic responses in the hippocampus. SHR rats were more sensitive to the F60 diet, suggesting that hypertension along with a high-fructose intake result in a more pronounced disorder compared to hypertriacylglycerolemia. This work wants to draw attention to fructose-induced health risks associated with hypertension.

Combination Therapy of Carnosic Acid and Methotrexate Effectively Suppressed the Inflammatory Markers and Oxidative Stress in Experimental Arthritis.

BACKGROUND: Combination therapy with methotrexate (MTX) is the most common therapeutic strategy used for the treatment of patients with rheumatoid arthritis (RA). In this study, we combined the natural compound carnosic acid (CA) with MTX to reduce inflammation and oxidative stress in adjuvant arthritis (AA). METHODS: AA was induced in 6-8 rats per group. MTX was administrated twice a week at a dose of 0.3 mg/kg b.w., while CA was administered daily at a dose of 100 mg/kg both in monotherapy and in combination with MTX. Plasma samples were collected on the 14th, 21st, and 28th day. Body weight and hind paw volume were measured once a week. RESULTS: We found that, mainly, the CA + MTX combination significantly reduced the hind paw swelling, the levels of IL-17A, MMP-9, and MCP-1 in plasma, and GGT activity in joint homogenates. The mRNA expression of HO-1, catalase, and IL-1ß in the liver were significantly improved by CA + MTX only. Our results indicate that adding CA to MTX treatment could be a good therapeutic option for patients suffering from RA. CONCLUSIONS: The addition of CA to methotrexate treatment significantly improved its efficacy in decreasing the development of AA by inhibiting the markers of inflammation and oxidative stress.

Enhanced Anti-Inflammatory Effect of the Combination of Lactiplantibacillus plantarum LS/07 with Methotrexate Compared to Their Monotherapies Studied in Experimental Arthritis.

The gut microbiome (GM) of rheumatic arthritis (RA) patients is often altered in composition and function. Moreover, methotrexate (MTX), one of the most frequently used disease-modifying antirheumatic drugs, is known to negatively affect GM composition. The modulation of immune system activity is one of the therapeutic benefits of probiotics. The aim of the current investigation was to determine the impact of MTX therapy combined with one of the Lactobacillus strains, Lactoplantibacillus plantarum LS/07 (LB), on adjuvant arthritis (AA) in rats. Methods focused on biometric and inflammatory parameters in AA, particularly on plasmatic levels of IL-17A, MMP-9, and MCP-1, and the activities of gamma-glutamyl transferase in the spleen and joints were applied. Enhancing the effect of MTX, LB positively influenced all biometric and inflammatory parameters. The findings of the present study may be of help in proposing novel therapeutic strategies for RA patients.

Bioflavonoid Robinin from Astragalus falcatus Lam. Mildly Improves the Effect of Metothrexate in Rats with Adjuvant Arthritis.

Anti-inflammatory potential of orally administrated bioflavonoid-robinin, active sub-stance of original drug Flaroninum™ (FL), was investigated in the combination with methotrexate (MTX) and in monotherapy in rats suffering from adjuvant-induced arthritis (AA). Robinin (kaempferol-3-O-robinoside-7-O-rhamnoside) was isolated from the aerial parts of Astragalus falcatus Lam. The monotherapy with robinin was not efficient in alleviating symptoms of AA. The combination of MTX with robinin was similarly active as MTX alone in reducing the hind paw volume and change of body weight during the whole experiment. The combination, however, reduced plasma levels of Interleukin-17Aand activity of gamma-glutamyl transferase in joint more efficiently then MTX alone. Our results demonstrate that the novel combination of robinin and MTX mildly improved the reduction of inflammation in experimental arthritis.

The effects of eggshell calcium (Biomin H® ) and its combinations with alfacalcidol (1α-hydroxyvitamin D3) and menaquinone-7 (vitamin K2) on ovariectomy-induced bone loss in a rat model of osteoporosis.

The purpose of this study was to investigate the impact of eggshell calcium (Biomin H® dietary supplement) and its combinations with alfacalcidol (1α-hydroxyvitamin D3 ) and menaquinone-7 (vitamin K2 ) on ovariectomy-induced bone loss in rats. Adult female rats (n = 48) were divided into 6 groups of 8 individuals each: sham-operated rats (SHAM); ovariectomized (OVX) rats untreated; OVX rats treated with Biomin H® (BIO); OVX rats simultaneously receiving Biomin H® , vitamin D3 (BIO + D3 ); OVX rats simultaneously treated with Biomin H® , vitamin K2 (BIO + K2 ) and OVX rats treated with Biomin H® , vitamin D3 , vitamin K2 (BIO + D3  + K2 ) during 8 weeks. Biochemical parameters, bone mineral density (BMD), bone mineral content (BMC) and femoral bone microstructure were determined. Plasma calcium and phosphate were increased in BIO + D3 and BIO + D3  + K2 groups as compared to OVX. Alkaline phosphatase was elevated in OVX, BIO versus SHAM, BIO + D3  + K2 groups. When compared to OVX group, decreased urine deoxypyridinoline was observed in all treated groups and femoral BMD, BMC were higher in BIO, BIO + D3 , BIO + D3  + K2 groups. The BIO + K2 rats had similar densitometrical values than OVX individuals. Microcomputed tomography revealed increased trabecular relative bone volume (due to an increase in trabecular number) in BIO + D3 , BIO + D3  + K2 as compared to OVX. The higher relative bone volume in BIO + D3 , BIO + D3  + K2 groups was also accompanied by an increase in bone surface. In the cortical bone, an enhanced periosteal bone apposition was identified in BIO, BIO + D3 , BIO + K2 , BIO + D3  + K2 groups. The rats from BIO + D3  + K2 group had a higher area of primary osteon's vascular canals. In BIO + D3 , BIO + K2 , BIO + D3  + K2 groups, an increased area of secondary osteons was determined in comparison with OVX. Our results indicate the beneficial effect of triple application of Biomin H® , vitamin D3 , vitamin K2 , as well as simultaneous administration of Biomin H® , vitamin D3 on the inhibition of ovariectomy-induced bone loss in a rat model of osteoporosis.

Skin wound healing with composite biomembranes loaded by tiopronin or captopril.

Novel wound dressings composed of chitosan (Ch) and hyaluronan (HA) loaded with tiopronin or captopril as antiinflammatory drugs were prepared. Composite biomembranes were examined in skin wounds of ischemic rabbits with the aim to accelerate the process of healing. The results proved that the biomembranes composed of Ch/HA/tiopronin or Ch/HA/captopril facilitated healing of skin wounds compared to untreated animals and animals treated with Ch/HA membranes. These results were confirmed by histology. Cu(II) ions and ascorbate-induced high-molar-mass HA degradation by means of rotational viscometry was performed and the ability of the both drugs to scavenge reactive oxygen species was evaluated. The results showed that captopril as well as tiopronin decreased the rate of HA degradation exclusively at higher concentrations.

Impact of Ergothioneine, Hercynine, and Histidine on Oxidative Degradation of Hyaluronan and Wound Healing.

A high-molecular weight hyaluronan is oxidatively degraded by Cu(II) ions and ascorbate-the so called Weissberger biogenic oxidative system-which is one of the most potent generators of reactive oxygen species, namely •OH radicals. Ergothioneine, hercynine, or histidine were loaded into chitosan/hyaluronan composite membranes to examine their effect on skin wound healing in ischemic rabbits. We also explored the ability of ergothioneine, hercynine, or histidine to inhibit hyaluronan degradation. Rotational viscometry showed that ergothioneine decreased the degree of hyaluronan radical degradation in a dose-dependent manner. While histidine was shown to be potent in scavenging •OH radicals, however, hercynine was ineffective. In vivo results showed that the addition of each investigated agent to chitosan/hyaluronan membranes contributed to a more potent treatment of ischemic skin wounds in rabbits compared to untreated animals and animals treated only with chitosan/hyaluronan membranes.

Triglyceride-lowering effect of the aldose reductase inhibitor cemtirestat-another factor that may contribute to attenuation of symptoms of peripheral neuropathy in STZ-diabetic rats.

Hyperglycemia is considered a key risk factor for development of diabetic complications including neuropathy. There is strong scientific evidence showing a primary role of aldose reductase, the first enzyme of the polyol pathway, in the cascade of metabolic imbalances responsible for the detrimental effects of hyperglycemia. Aldose reductase is thus considered a significant drug target. We investigated the effects of cemtirestat, a novel aldose reductase inhibitor, in the streptozotocin-induced rat model of uncontrolled type 1 diabetes in a 4-month experiment. Markedly increased sorbitol levels were recorded in the erythrocytes and the sciatic nerve of diabetic animals. Osmotic fragility of red blood cells was increased in diabetic animals. Indices of thermal hypoalgesia were significantly increased in diabetic rats. Tactile allodynia, recorded in diabetic animals in the early stages, turned to mechanical hypoalgesia by the end of the experiment. Treatment of diabetic animals with cemtirestat (i) reduced plasma triglycerides and TBAR levels; (ii) did not affect the values of HbA1c and body weights; (iii) reversed erythrocyte sorbitol accumulation to near control values, while sorbitol in the sciatic nerve was not affected; (iv) ameliorated indices of the erythrocyte osmotic fragility; and (v) attenuated the symptoms of peripheral neuropathy more significantly in the middle of the experiment than at the end of the treatment. Taking into account the lipid metabolism as an interesting molecular target for prevention or treatment of diabetic peripheral neuropathy, the triglyceride-lowering effect of cemtirestat should be considered in future studies. The most feasible mechanisms of triglyceride-lowering action of cemtirestat were suggested.

Natural substance rutin versus standard drug atorvastatin in a treatment of metabolic syndrome-like condition.

BACKGROUND: Metabolic syndrome is a cluster of metabolic risk factors. The clear causes of its development are not known yet and there is no comprehensive treatment of this disease. There is a trend to use natural substances in the treatment of various diseases, but their effects need to be well explored. We decided to test effect of rutin compared to the effect of the standard drug atorvastatin. METHODS: As a model of metabolic syndrome we used males of hypertriacylglycerolemic rats in combination with high-fat-high-fructose diet. Rutin (100 mg/kg) and atorvastatin (50 mg/kg) were administered orally daily for 5 weeks. RESULTS: We determined biochemical parameters from blood: HDL-cholesterol, LDL-cholesterol, total cholesterol, triacylglycerols. Relaxation and contraction response of aorta was measured to determine vessel dysfunctions and possible predisposition to cardiovascular disease. The negative influence on cognitive functions could be associated with the development of metabolic cognitive syndrome. Therefore we aimed to monitor spatial memory by Morris water maze test. Both rutin and atorvastatin had a tendency to decrease levels of serum triacylglycerols, but only atorvastatin significantly reduced levels od LDL-cholesterol and increased HDL-cholesterol levels. Both compounds significantly reduced the phenylephrine-induced contractile response of the aorta and improved the relaxation response. Further, treated animals learned better compared to untreated rats in the Morris water maze. CONCLUSION: Based on our results we can assume that atorvastatin and rutin had positive effect on spatial memory and vessel reactivity. Atorvastatin optimized lipid profile of blood serum.

Cornelian cherry fruit improves glycaemia and manifestations of diabetes in obese Zucker diabetic fatty rats.

Cornelian cherry (Cornus mas L.) was in the past frequently used in Slovak Republic; meanwhile fell into oblivion despite the fact that it is known as antidiabetic supplement. However, there is no research investigated its effect on animal model of Diabetes mellitus (DM) 2 type as it is Zucker diabetic fatty (ZDF) rats. Thus, the aim of this study was to determine the effect of C. mas fruit given orally on the development of DM symptoms in ZDF rats. In the experiment male ZDF rats (fa/fa) and their age-matched non-diabetic lean controls (fa/+) were used aged 12 weeks. Male ZDF rats were administered C. mas in two doses (500 and 1000 mg/kg body weight) using a gastric gavage for 10 weeks. One group of diabetic animals served as positive control and received only distilled water. We found significant decrease of glucose level after oral administration of C. mas in dose of 1000 mg/kg bw in pre-diabetic state of animals (until 7th week of the experiment) and significant restriction of water intake in both C. mas groups against the diabetic control. We presume that the higher dose of Cornelian cherry could be beneficial and helpful in prevention of diabetic symptoms when consumed regularly in young animals.

3-Mercapto-5H-1,2,4-Triazino[5,6-b]Indole-5-Acetic Acid (Cemtirestat) Alleviates Symptoms of Peripheral Diabetic Neuropathy in Zucker Diabetic Fatty (ZDF) Rats: A Role of Aldose Reductase.

Peripheral neuropathy is the most prevalent chronic complication of diabetes mellitus. Good glycemic control can delay the appearance of neuropathic symptoms in diabetic patients but it is not sufficient to prevent or cure the disease. Therefore therapeutic approaches should focus on attenuation of pathogenetic mechanisms responsible for the nerve injury. Considering the role of polyol pathway in the etiology of diabetic neuropathy, we evaluated the effect of a novel efficient and selective aldose reductase inhibitor, 3-mercapto-5H-1,2,4-triazino[5,6-b]indole-5-acetic acid (cemtirestat), on symptoms of diabetic peripheral neuropathy in Zucker Diabetic Fatty (ZDF) rats. Since the age of 5 months, male ZDF rats were orally administered cemtirestat, 2.5 and 7.5 mg/kg/day, for two following months. Thermal hypoalgesia was evaluated by tail flick and hot plate tests. Tactile allodynia was determined by a von Frey flexible filament test. Two-month treatment of ZDF rats with cemtirestat (i) did not affect physical and glycemic status of the animals; (ii) partially inhibited sorbitol accumulation in red blood cells and the sciatic nerve; (iii) markedly decreased plasma levels of thiobarbituric acid reactive substances; (iv) normalized symptoms of peripheral neuropathy with high significance. The presented findings indicate that inhibition of aldose reductase by cemtirestat is not solely responsible for the recorded improvement of the behavioral responses. In future studies, potential effects of cemtirestat on consequences of diabetes that are not exclusively dependent on glucose metabolism via polyol pathway should be taken into consideration.

MitoQ Loaded Chitosan-Hyaluronan Composite Membranes for Wound Healing.

Two self-associating biopolymers, namely chitosan (Ch) and a high-molar-mass hyaluronan (HA), were used to prepare membranes with the aim to protect and to enhance the healing of injured skin. A mitochondrially-targeted antioxidant-MitoQ-was incorporated into the mixture of biopolymers prior to their self-association. These three-component membranes were evaluated in detail utilising surface roughness measurements, contact angle measurements, hemocompatibility, and thrombogenicity analyses. Furthermore, in vivo application of Ch/HA/MitoQ membranes was assessed on injured rabbit and rat skin utilizing histological methods. The results showed that the prepared thrombogenic Ch/HA/MitoQ membranes had higher roughness, which allowed for greater surface area for tissue membrane interaction during the healing processes, and lower cytotoxicity levels than controls. MitoQ-loaded composite membranes displayed superior healing properties in these animal models compared to control membranes.

Ferulaldehyde Improves the Effect of Methotrexate in Experimental Arthritis.

Methotrexate (MTX) is still the gold standard for treatment of rheumatoid arthritis (RA). The therapeutic efficacy of low-dose of MTX can be increased by its combination with a natural substance, ferulaldehyde (FRA). The aim of this study was to evaluate the effect FRA and MTX administered alone or in combination in adjuvant arthritis. The disease was induced to Lewis male rats by intradermal injection, which contains a suspension of heat-inactivated Mycobacterium butyricum in incomplete Freund's adjuvant. The experiment of 28 days included: healthy animals, arthritic animals, arthritic animals with administration of FRA at the oral daily dose of 15 mg/kg, arthritic animals with administration of MTX at the oral dose of 0.3 mg/kg twice a week, and arthritic animals administered with FRA and MTX. FRA in monotherapy decreased significantly only the level of interleukin-1ß (IL-1ß) and matrix metalloproteinase-9 in plasma. Combination of FRA and low-dose MTX was more effective than MTX alone when comparing body weight, hind paw volume, arthritic score, plasmatic levels of IL-1ß, activity of γ-glutamyl transferase, and relative mRNA expression of IL-1ß in the spleen. Therefore, the combination treatment was the most effective. The obtained results are interesting for future possible innovative therapy of patients with RA.

Cryptococcus laurentii extracellular biopolymer production for application in wound management.

Cryptococcus laurentii growth and extracellular polysaccharide (EPS) production in bioreactor were studied. Biomass yield 14.3 g/L and EPS synthesis 4.3 g/L in 144 h of submerged cultivation were achieved. EPS synthesis and cell growth had different optima. For EPS formation, pH 3, 25 °C and low aeration (1 % < pO2 < 10 %) were advantageous, while cell growth optimum was at pH 6, 20 °C, and high aeration (pO2 > 30 %). As medium pH changed from pH 3 to pH 6, glucuronic acid (GluAc) content in EPS increased, while galactose, xylose, and glucose decreased. Twenty-five degrees Celsius was optimal for GluAc containing polysaccharide synthesis, while lower temperature (15 °C) increased glucose content in EPS. Aeration intensity and time of cultivation had little effect on EPS composition. Molecular mass distribution of raw C. laurentii EPS was determined by SEC-MALS as 1.352. The row EPS was composed of acidic glucuronoxylomannan for more than 85 %. In the in vivo experiments, EPS significantly improved excisional wound healing in healthy rats. The results suggest that C. laurentii EPS is a promising biotechnological product and an advanced material for application in wound management.

Chondrogenic differentiation of mesenchymal stem cells in a hydrogel system based on an enzymatically crosslinked tyramine derivative of hyaluronan.

Hyaluronan-based tissue substitutes are promising materials in cartilage reconstruction surgery. Herein, the chondrogenesis of human mesenchymal stem cells (MSC) in a hydrogel based on a tyramine derivative of hyaluronan crosslinked by hydrogen peroxidase (HA-TA) was evaluated. Human MSC seeded in the scaffold were incubated in standard chondrogenic medium and medium enriched with bone morphogenetic protein-6 (BMP6). Cell viability, the gene expression of selected markers (collagen type II, aggrecan, SOX9, collagen type X, and osteopontin), and the histological characteristics were examined during three weeks of in vitro cultivation. The tissue reaction of both unseeded and MSC seeded HA-TA scaffolds were tested in vivo after subcutaneous application in rats for 12 weeks. The data showed that cells resisted the process of crosslinking and remained viable for the whole time while exhibiting changes in cell organization. Human MSC cultivated in HA-TA hydrogel expressed genes of both chondrogenic and osteogenic differentiation and the addition of BMP6 revealed a tendency to potentiate both processes. Histological analysis of HA-TA in vivo implants did not reveal a chronic inflammatory reaction. In both cases, in vivo HA-TA implants were continuously degraded and MSC-seeded hydrogels tended to form clusters similar to in vitro samples. In conclusion, MSC chondrogenic differentiation may proceed in a HA-TA scaffold that is biocompatible. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 3523-3530, 2014.

Preclinical evaluation of wound treatment with hyaluronan-iodine hydrogel.

UNLABELLED: A mixture of hyaluronan and iodine complex KI3 (Hyiodine®) has been developed to support wound healing. In this study, the effect of hyaluronan, KI3, and their combination on progression of wound healing in excision skin wounds in rats was determined. METHODS: To evaluate the possible toxic effects of iodine after Hyiodine application for wound healing, iodine systemic absorption from the Hyiodine-treat- ed wounds and its distribution profile were quantitatively described and compared after intravenous iodide administration using 131I as a radiolabel. RESULTS: Treatment of rats with Hyiodine resulted in an enhancement of wound healing, proved by greater degrees of wound contraction and reduction in mean wound healing time compared to other treated rats on hours 1, 2, 24, and 216. Even if wound therapy with Hyiodine resulted in systemic iodine uptake, the estimated iodine absorbed dose in human therapy is tolerable, and the theoretical risk of its systemic toxic effects is minimal. CONCLUSION: A hyaluronan-iodine hydrogel has a great potential for effective treatment of wounds. .

Treatment of rat adjuvant arthritis with flavonoid (Detralex), methotrexate, and their combination.

In both adjuvant arthritis and rheumatoid arthritis, edema and inflammation appear in synovial joints. Edema or effusion reflects an imbalance in lymph dynamics. Purified micronized flavonoid fraction is mainly used in the treatment of chronic venous insufficiency. This compound improves lymphatic drainage with a significant increase in lymphatic flow and lymphatic pulsality. It is suggested that the beneficial effect of purified micronized flavonoid fraction may be involved in the treatment of adjuvant arthritis in rats. In this study treatment of adjuvant arthritis in rats with Detralex, methotrexate, and their combination were evaluated. Groups of rats with adjuvant arthritis were treated with methotrexate (0.6 mg/kg/week), Detralex (20 mg/kg/day), and their combination for 50 days from adjuvant application. Hind paw swelling, arthrogram scores, serum albumin level, serum nitrite/nitrate concentrations, and whole-body mineral density were evaluated as markers of inflammation and destructive changes associated with arthritis. Long-term prophylactic treatment with low-dose methotrexate significantly inhibited the markers of both inflammation and arthritis. Detralex administered alone slightly decreased both the hind paw swelling and the arthritic score. Other inflammatory and arthritic markers were not significantly influenced. However, Detralex combined with methotrexate markedly potentiated the beneficial effects of methotrexate, which resulted in a more significant reduction in hind paw swelling, arthritic scores, and serum concentrations of nitrite/nitrate. Interestingly, the arthritis-induced decrease of bone mineral density in AA rats was significantly lower only in the group treated with the combination of Detralex and methotrexate. Our results indicate that Detralex increased the therapeutic efficacy of methotrexate basal treatment in AA. We suggest that this may be related to the beneficial effect of Detralex on microcirculation, especially on venules and lymphatic vessels.

Methotrexate treatment ameliorated testicular suppression and anorexia related leptin reduction in rats with adjuvant arthritis.

Methotrexate (MTX) has been frequently used in the treatment of rheumatoid arthritis (RA). However, its action on arthritis associated male hypogonadism, or anorexia related low leptin production has not yet been studied. The well-established model of human RA is rat adjuvant-induced arthritis (AA). In the present series we aimed at the evaluation of the effects of MTX on AA induced inflammatory parameters, testosterone suppression, and anorexia associated lowered leptin release. AA was induced in male Lewis rats by intradermal injection of heat killed Mycobacterium butyricum in incomplete Freund's adjuvant in the base of the tail. Arthritic rats were treated with two doses of MTX: 0.3 and 0.5 mg/kg twice a week orally for the period of 28 days. The evaluated parameters were body mass, hind-paw swelling, arthrogram scores, serum albumin, total testosterone and leptin on days 14, 21 and 28 of AA. MTX treatment ameliorated all parameters studied dose dependently. Higher dose of MTX induced a significant reduction in the hind-paw swelling, arthritic score, and an increase in serum albumin in all examined time intervals of AA. This dose also significantly improved the suppressed testosterone and leptin levels found in arthritic rats. Prophylactic MTX treatment of rats with AA improved all inflammatory and arthritic parameters studied indicating its clear anti-inflammatory effects. The significant improvement of testosterone and leptin shows beneficial effects of MTX on reproduction and anorexia related leptin reduction during chronic AA.

Effects of purified micronized flavonoid fraction (Detralex) on prophylactic treatment of adjuvant arthritis with methotrexate in rats.

BACKGROUND: In both adjuvant arthritis and rheumatoid arthritis, edema and inflammation appear in synovial joints. Edema or effusion reflects an imbalance in lymph dynamics. Purified micronized flavonoid fraction is mainly used in the treatment of chronic venous insufficiency. This compound improves lymphatic drainage with a signicant increase in lymphatic flow and lymphatic pulsality. It is suggested that the beneficial effect of purified micronized flavonoid fraction may be involved in the treatment of adjuvant arthritis in rats. OBJECTIVES: To evaluate the effect of Detralex on methotrexate prophylactic treatment of adjuvant arthritis in rats. METHODS: Groups of rats with adjuvant arthritis were treated with methotrexate (0.6 mg/kg/week), Detralex (20 mg/kg/day) and their combination for 50 days from adjuvant application. Hind paw swelling, arthrogram scores, serum albumin level, serum nitrite/nitrate concentrations and whole body mineral density were evaluated as markers of inflammation and destructive changes associated with arthritis. RESULTS: Long-term prophylactic treatment with low dose methotrexate significantly inhibited the markers of both inflammation and arthritis. Detralex administered alone slightly decreased both the hind paw swelling and the arthritic score. Other inflammatory and arthritic markers were not significantly influenced. However, Detralex combined with methotrexate markedly potentiated the beneficial effects of methotrexate, which resulted in a more significant reduction in hind paw swelling, arthritic scores, and serum concentrations of nitrite/nitrate. Interestingly, the arthritis-induced decrease of BMD in AA rats was significantly lower only in the group treated with the combination of Detralex+methotrexate. CONCLUSION: Detralex increased the therapeutic efficacy of methotrexate basal treatment in AA. We suggest that this may be related to the beneficial effect of Detralex on microcirculation, especially on venules and lymphatic vessels.