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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.

Bee products are used in therapy of various diseases many years. As diabetes mellitus type 2 (DM2T) became a pandemic disease the searching for its treatment is necessary. In the literature there is evidence about beneficial properties of bee products in treatment of DM, but not about a specific bee product – bee bread. The aim of this study was to analyse the effect of bee bread on DM complications in the suitable animal model for research in diabetes mellitus type 2 Zucker diabetic fatty (ZDF) rats. Male ZDF rats (diabetic, n = 20) and their lean controls (non-diabetic, n = 10) in the age of 3 months were used in the experiment. Animals were provided with water and diet (complete mixture for rats - KKZ-P/M, 10 MJ kg−1) on ad libitum base. Rats were divided into three groups as follows: lean untreated rats (Lean), obese diabetic rats without any additives (control, C, given distilled water daily using sterile oral rodent gavage) and bee bread group (bee bread in a dose 700 mg kg−1 of body weight daily using sterile oral rodent gavage). Treatment of the diabetic animals with bee bred caused significant decrease of glucose from the 3rd till the 9th week of the experiment when compared to the diabetic control. We found also significantly lower intake of water in bee bread group from the 4th till the 6th week of the experiment against diabetic control. In conclusion, bee bread beneficially affected glucose metabolism and water intake in diabetic rats, thus shows the potential for the treatment of hyperglycaemia and other disorders in diabetes.

The purpose of this study was to investigate the impact of eggshell calcium (Biomin H

The appropriate animal model of diabetes mellitus type 2 is Zucker diabetic fatty (ZDF) rats. The goal of this study was to analyse the effect of chronic high-energy diet on diabetes mellitus (DM) complications in ZDF rats. Male ZDF rats (n = 20) and their lean controls (non-diabetic, n = 10) in the age of 3 months were involved in the experiment. Rats were provided with water and diet on ad libitum base. Animals were divided into three groups as follows: lean untreated rats (C) fed by KKZ-P/M (10 MJ/kg), obese rats fed by KKZ-P/M (10 MJ/kg, E1) and obese rats fed by enriched high energy diet (E2, enriched KKZ-P/M, 20 MJ/kg). Glucose, ketones levels, the consumption of feed, water and the live weight was measured weekly during the whole experiment. At the end of the experiment rats were anesthetized and selected haematological parameters were measured. ZDF rats in E1 and E2 group developed obesity, hyperglycaemia, non-insulin dependent diabetes, aggravations in haematological parameters and accumulation of sorbitol in sciatic nerve and lens of rats. High-energy diet immediately induced hyperglycaemia followed by accelerating the secondary symptoms of diabetes complications expressed by disturbed haematology parameters. High-energy diet caused ketoacidosis what meant two cases of death. Extended research on diabetes is needed.

Combination of methotrexate (MTX) with hydroxychloroquine (HCQ) is one of the most effective therapies used for treatment of rheumatoid arthritis. The aim of the study was to more complexly examine the effect of combination HCQ and MTX on disease progression in arthritic rats focusing on inflammatory markers. Adjuvant arthritis (AA) was induced by intradermal injection of Mycobacterium butyricum in incomplete Freund´s adjuvant in Lewis rats and the disease was monitored 28 days. The experiment included healthy animals, control AA animals, AA animals with administration of HCQ (40 mg/kg b.w. daily, p.o.), AA animals with administration of MTX (0.4 mg/kg b.w. twice a week, p.o.), and AA animals treated with the combination of HCQ + MTX. HCQ in monotherapy reduced only the plasmatic level of IL-1β (day 28). On day 14, MTX + HCQ significantly decreased the plasmatic levels of IL-1β, IL-17 and combination was more effective than MTX in reduction of IL-17. On day 28, combination significantly decreased the plasmatic levels of CRP and IL-1β and it was more effective than MTX in reduction of IL-1β. Expression of IL-1β and inducible NO-synthase total mRNA in lung were most effectively reduced by MTX + HCQ. Combination of MTX + HCQ reduced hind paw volume, arthritic score and increased body mass of animals very effectively (days 14, 21, and 28). In summary, MTX + HCQ were much better than MTX in correcting inflammatory and clinical markers. This study showed in more details that the anti-arthritic effect of MTX + HCQ is particularly due to reduction of the levels of inflammatory cytokines.

Background Green tea catechins are widely employed as dietary supplements to restore the oxidative state in adjuvant-induced arthritic rats, a model for human rheumatoid arthritis. Methotrexate is the first line antirheumatic drug. The present study aimed to investigate if a decaffeinated extract (GreenSelect®, GS) of tea polyphenols can ameliorate methotrexate treatment in rat adjuvant arthritis. Methods The study lasted 28 days and included healthy animals administered with GS (daily dose of 200 mg/kg), untreated arthritic rats and arthritic rats treated with: GS, methotrexate in single treatment or in combination with GS. Arthritic score and changes in body weigh were measured during the treatment while inflammatory markers (monocyte chemotactic protein-1, IL-17 and inducible NO-synthase mRNA) and biochemical parameters (gamma-glutamyltransferase and heme oxygenase-1) at the end of the treatment. Results The association between GS and methotrexate was less efficient in ameliorating the arthritic score compared to methotrexate alone. GS did not improve the inflammatory and biochemical markers except for monocyte chemotactic protein-1 and negatively affected the body weight. GS did not increase the plasma antioxidant capacity, suggesting a pro-oxidant effect. Conclusions The results suggest that long-term administration of GS may inhibit the therapeutic action of methotrexate in arthritic rats.

In the development of rheumatoid arthritis (RA) and its experimental model rat adjuvant arthritis (AA), immunological and inflammatory pathological changes are involved. Although a number of synthetic medications are used to treat RA, none of the current regimens are completely safe. Dietary phytochemicals, mainly polyphenols, are some of the most promising natural sources for developing novel functional foods and drugs with improved efficacy and safety. Several important clinical and preclinical in vivo and in vitro studies support the use of natural compounds as complementary intervention to help manage RA. Results from our three experiments with phytochemicals—pinosylvin, N-feruloylserotonin, and ferulaldehyde—administered alone or in combination with methotrexate in the AA model have shown promise for development of novel agents for RA treatment. The latest findings from the literature about the antiinflammatory activity of polyphenols as well as from our experiments are summarized in this chapter.

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.

Novel therapies for rheumatoid arthritis also include the use of naturally occurring compounds possessing antioxidant properties. In the present work, the effects of oral administration of quercetin were investigated in a rat model of adjuvant arthritis. Arthritis was induced by a single intradermal injection of heat-inactivated Mycobacterium butyricum in incomplete Freund's adjuvant. The experimental groups were treated with an oral daily dose of 150 mg/kg b.w. of quercetin for 28 days. Results indicated that quercetin was able to ameliorate all markers of inflammation and oxidative stress measured. Quercetin lowered levels of interleukin-1β, C-reactive protein, and monocyte chemotactic protein-1 and restored plasma antioxidant capacity. In addition, quercetin inhibited the enzymatic activity of pro-inflammatory 12/15-lipoxygenase in lung and liver and increased the expression of heme oxygenase-1 in joint and lung of arthritic rats. Finally, quercetin inhibited the 2-fold increase of NF-қB activity observed in lung, liver and joint after induction of 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-1β (IL-1β) 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-1β, activity of γ-glutamyl transferase, and relative mRNA expression of IL-1β 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.