Royal jelly (RJ) is a gelatinous substance that bees produce to feed bees and queen bees. It’s frequently sold as a dietary supplement to treat a variety of physical ailments and chronic diseases. While it has long been used in traditional medicine, its applications in Western medicine remain controversial. The inhibitory effect of royal jelly on osteoarthritis was investigated in primary cultured rat cartilage cells and monosodium-iodoacetate (MIA)-induced arthritis rat model 10-hydroxy-2-decenoic acid (10-HAD) is the main fatty acid present in RJ. Among the criteria for RJ quality analysis, 10-HAD content has been proposed as a freshness parameter. We investigated the effect of RJ on the improvement of osteoarthritis on SD rats and they were divided into five groups. In this study, we examined the effect of enzymatic royal jelly (ERJ) administration on osteoarthritis. To determine the antiinflammatory effects of RJ, tumor necrosis factor alpha (TNF-α) and Interleukin-6 (IL-6) expression were measured after lipopolysaccharide (LPS) activation in RAW 264.7 cells. In in vivo animal study, osteoarthritis was induced by intra-articular injection of MIA into knee joints of rats. As a results, ERJ showed that TNF-α and IL-6 levels were decreased by ERJ treatment in a dosedependent manner. In conclusion, ERJ extract was able to inhibit articular cartilage degeneration by preventing extracellular matrix degradation and cartilage cell damage. It was considered that ERJ extract may be a potential therapeutic treatment for degenerative osteoarthritis.
The purpose of this study was to examine the characteristics of acetaminophen (APAP)-induced liver damage, using fluorescence bioimaging, serum biochemistry, and histopathology. At six weeks of age, eighteen mice were divided into three groups as group 1 (G1) as control, group 2 (G2) as fluorescence probe control and group 3 (G3) as APAP-treated. G3 mice were orally treated with APAP (800 mg/kg b.w.), while G1 and G2 mice were treated with 0.9% saline. Twenty-two hours after APAP treatment, G2 and G3 mice were intravenously treated with Annexin-Vivo 750 as probe, while G1 mice were treated with saline. Fluorescence bioimaging was performed at two hours after probe treatment. The mice were sacrificed and serum levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and lactate dehydrogenase were analyzed. Liver damage was examined by hematoxylin and eosin (H&E) staining and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. In vivo bioimaging, fluorescence intensity of the region of interest (ROI) was significantly increased in the livers of G2 and G3 mice compared with those in G1 mice (p<0.05 and p<0.01). In addition, ex vivo bioimaging confirmed that the fluorescence intensity of the ROI was significantly increased in the livers of G2 and G3 mice compared with those in G1 mice (p<0.05 and p<0.01). All examined serum parameters of G3 were significantly increased compared with G1 and G2 (p<0.05 and p<0.01). H&E examination showed acute hepatic cell necrosis in the livers of G3 mice, while there was no cell death in the livers of G1 and G2 mice. TUNEL staining also showed many cell death features in G3 mice, whereas no pathological findings were shown in G1 or G2 mice. In summary, fluorescence bioimaging showed the possibility of cell death detection in the livers of mice treated with APAP, and this was corroborated by serum chemistry and histopathological examination.
Lysyl oxidase-variant 2 (LOX-v2) is a novel variant of lysyl oxidase (LOX) that functions as an amine oxidase for the formation of lysine-mediated crosslinks found in collagen and elastin fibrils. In addition to the amine oxidase activity in the extracellular matrix, several novel functions, such as tumor suppression, tumor progression, chemotaxis, cellular senescence, and modification of histones, have been assigned to LOX. In recent years, it has been reported that LOX is also present in nuclear locations, suggesting a novel functional role of LOX in the nucleus. To test the amine oxidase activity of LOX and LOX-v2 to nuclear histone proteins, we expressed and purified LOX and LOX-v2 as recombinant forms and then assessed the amine oxidase activity toward histone H2A in in vitro peroxidase-coupled fluorometric assays. Both LOX and LOX-v2 proteins showed significant levels of amine oxidase activity toward histone H2A in a β -aminopropionitrile-inhibitable manner. In immunofluorescence staining after ectopic expression in cultured cells, LOX was observed in the perinuclear, cytoplasmic, and extracellular areas, whereas LOX-v2 was predominantly detected in the nucleoplasm with a punctuate pattern. These findings suggest that LOX-v2 may play a novel functional role in the nucleus through the amine oxidase activity to the nuclear histone proteins. Elucidation of the specific functional roles of LOX-v2, such as substrate specificity toward different types of nuclear proteins and detailed analysis on subnuclear localization, will provide a significant clue in understanding the diverse functional roles currently assigned to a single enzyme, LOX.
In previous studies, we found the production of antibodies against cross-reactive bovine serum albumin (BSA) in D-galactose (D-gal) induced aging mouse models. We performed immunoblot analysis with mouse tissue lysates to investigate the changes in the overall autoantibody production in this animal model. And we were able to see the possibility of altering the activity of mouse natural antibodies in this process. In this study, we examined changes in existing natural antibodies in a D-gal-induced aging mouse model. Serum samples were collected from 3-week-old mice (3w), 13-week-old mice (13w), and 13-week-old mice that were treated with D-gal for 6 weeks (13wDG), beginning at the age of 8 weeks. Levels of immunoglobulins (IgM, IgG, and IgA) were quantitatively analyzed in serum samples. Tissue samples were obtained from skin, spleen, and ovary for Western blotting analyses. Natural antibody activity was examined by enzyme-linked immunosorbent assay analyses of anti- double-stranded DNA (dsDNA) antibody. Western blotting analyses using mouse tissue lysates showed that several protein bands detected by serum antibodies from 3w mice became increasingly thicker when detection was performed with serum samples from 13w and 13wDG mice, indicating quantitative increases in levels of natural antibodies. Relative amounts of total IgG, IgM, and IgA immunoglobulins sequentially increased in serum samples from 3w, 13w, and 13wDG mice. A similar tendency was observed regarding the levels of IgM and IgG antibodies against dsDNA. These results indicate increased levels of natural antibodies in the D-gal-induced aging mouse model. Therefore, this animal model could be useful for future natural antibody research.
Artemisia annua (AA) is a well-known as a source of antimalarial drug (artemisinin), which also has been traditionally used as an antipyretic and hemostatic agent in Korea and China. In preclinical effective study, a water extract of Artemisia annua (WEAA) ameliorated weight gain and hepatic lipid accumulation in high-fat diet-fed mice. The plasma levels of triglyceride, AST, and ALT were reduced in the WEAA-treated group. Based on these results, the safety of WEAA as a functional ingredient for liver health was evaluated in this repeated dose oral toxicity study before the clinical trial. Sprague- Dawley (SD) rats were treated by gavage with 20 times (1,000 mg/kg) more than the effective dose for 13 weeks. All rats had survived at the end of the study, and there were no changes indicating obviously abnormal clinical sign and behavior. The treatment of WEAA were also observed no obvious toxicities in the body weights, urine, hematological, serum biochemical, ophthalmic and histopathological examinations. Based on the results of this study, the NOAEL (no-observed-adverse-effect level) of WEAA in SD rats was estimated to be 1,000 mg/kg. In conclusion, WEAA could be used as a safe functional ingredient for the improvement of liver health in individuals with hepatic diseases including nonalcoholic steatohepatitis.