Harry Jung

Anti-diabetic Effect of the Soybean Extract Fermented by Bacillus subtilis MORI in db/db Mice

This study investigated the effects of soy bean extract fermented by Bacillus subtilis MORI (BTD-1) on blood glucose, hemoglobin A1c (HbA1c), plasma insulin, and pancreatic β islets in db/db mice. The BTD-1 (500 mg/kg) group showed significantly lower fasting blood glucose level (p<0.01) and postprandial 2 h blood glucose level (p<0.01) compared with the db control group. The BTD-1 (500 mg/kg) group showed significantly lower HbA1c level compared with the db control group (p<0.01). After 8 weeks of BTD-1 administration, the pancreatic islet architecture was preserved and the immunofluorescent intensities of insulin in BTD-1 (500mg/kg) group apparently increased compared to in the db control group. Plasma insulin levels were found to be significantly higher in the BTD-1 (500 mg/kg) group than in the db control group (p<0.05). In summary, our results suggest that BTD-1 has an anti-diabetes effect in a non-insulin dependent diabetes mellitus mouse model.

Subcellular localization of the transmembrane inner ear (Tmie) protein in a stable Tmie-expressing cell line

Mutations in the transmembrane inner ear (Tmie) gene, which encodes the Tmie protein, have been attributed to deafness autosomal recessive 6 (DFNB6), an autosomal nonsyndromic recessive hearing loss disorder. Although the Tmie gene was identified a few years ago, little is known about subcellular localization of the Tmie protein. In order to address this, we developed a stable cell line expressing Tmie protein. The expression of Myc-tagged Tmie protein was confirmed by Western blot analysis using an anti-Myc antibody and localization of the Tmie protein was confirmed by immunostaining, using the anti-Myc antibody as well as the anti-tmie antibody. Our study demonstrates that the Tmie protein is localized mostly in the cellular membrane and to a lesser extent in cytoplasm. These results suggest that our Tmie expressing stable cell line provides a suitable in vitro model to explore Tmie synthesis and functions.

Anti-hyperlipidemic effect of soybean extract fermented by Bacillus subtilis MORI in db/db mice

The purpose of this study was to investigate the anti-hyperlipidemic effect of soy bean extract solution fermented by Bacillus subtilis MORI (BTD-1E) in obese db/db mice. Eight-week-old male db/db mice were administered 33.3 mg/kg BTD-1E solution orally once a day for four weeks. The BTD-1E group showed significantly lower body weight compared with the db control group (P<0.05). The BTD-1E group showed significantly lower serum total cholesterol and LDL cholesterol levels compared with the db control group, respectively (P<0.05, P<0.01). The BTD-1E group showed significantly decreased liver weight relative to final body weight compared with the db control group (P<0.01). After four weeks of BTD-1E administration, lipid droplets in the liver were apparently decreased in the BTD-1E group compared to the db control group. In summary, our results suggest that BTD-1E has an anti-hyperlipidemic effect in the obese mouse model.

Rapid and efficient identification of the mouse leptin receptor mutation (C57BL/KsJ-db/db) by tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) analysis

The C57BLKS/J-Leprdb mouse has a point mutation in the leptin receptor gene and is one of the most useful animal model for non-insulin dependent diabetes mellitus in human. Since the homozygote of C57BLKS/J-Leprdb mouse is infertile, detection of point mutation in the leptin receptor gene is important for efficient maintaining strains as well as mass production of homozygotes. To develop a rapid and efficient genotyping method for C57BLKS/J-Leprdb mouse, the tetra-primer amplification-refractory mutation system polymerase chain reaction (ARMS-PCR) was used. The 407 and 199 bp PCR products were amplified from normal (+/+) mice; while the 407 and 268 bp PCR products were amplified from homozygotes (db/db) mice; and the 407, 268, and 199 bp PCR products were amplified from heterozygotes (db/+) mice. This result showed that the tetra-primer ARMS-PCR analysis by us is suitable to detect point mutation of the leptin receptor gene. Taken together, our method will dramatically reduce animal use for maintenance of strains as well as production of homozygote in the C57BLKS/J-Leprdb strains.

Anti-obesity Effects of the Stem Bark of Japanese Horse Chestnut (Aesculus turbinate) in 3T3-L1 Preadipocytes

The inhibitory effects of lipid accumulation on ethanol extract from stem bark of Japanese horse chestnut (JHC) were evaluated. Exposure to JHC extract (10–100 μg/mL) for a 72 h incubation period did not alter cell viability compared to the untreated control. JHC extract, with concentrations of 25, 50, and 100 μg/mL, inhibited lipid accumulation in 3T3-L1 adipocytes in a dose dependent manner. The expression of PPARγ and C/EBPα, important adpogenic key markers was significantly reduced when JHC extract was added to cells for 8 days compared with the untreated control group. These results suggest that JHC extract might be a potential therapeutic agent as a natural anti-obesity material.

Anti-adipogenic effects in 3T3-L1 cells of acetone extracts and fractions from Styrax japonica fruit

Reduction in reactive oxygen species (ROS) production facilitated by Styrax japonica (SJ) extracts and fractions for inhibition of obesity development through inhibition of adipogenesis was investigated. Treatment of cells with SJ extracts and fractions did not result in changes in cell viability. SJ extracts and fractions inhibited lipid accumulation in a dose-dependent manner during adipogenesis. Expressions of PPARγ and C/EBPα, key adipogenic markers, were decreased in cells treated with SJ extracts and fractions. Inhibitory effects of SJ extracts and fractions were the most powerful in the early stages of differentiation (days 0-2). Intracellular ROS production was decreased in cells treated with SJ extracts and fractions. SJ extracts and fractions can inhibit adipogenesis via reduced ROS levels, which involves down-regulation of adipogenic transcription factors.

Fagopyritol, a Derivative of D-chiro-inositol, Induces GLUT4 Translocation via Actin Filament Remodeling in L6-GLUT4myc Skeletal Muscle Cells

Insulin induces glucose transporter 4 (GLUT4) translocation to the muscle cell surface. As fagopyritol has insulin-like effects, the effects of fagopyritol on GLUT4 translocation and filamentous (F) actin remodeling in L6-GLUT4myc skeletal muscle cells were investigated. Fagopyritol significantly increased plasma membrane GLUT4 levels compared with the basal control in L6-GLUT4myc myoblast cells. Phosphatidylinositol (PI) 3-kinase inhibitor (LY294002) treatment prevented GLUT4 translocation to the plasma membrane in the myoblasts. Fagopyritol treatment apparently stimulates F-actin remodeling in myoblasts. In addition, fagopyritol treatment induced GLUT4 translocation and F-actin remodeling in myotubes. Taken together, these results suggest that fagopyritol promotes GLUT4 translocation and F-actin remodeling by activating the PI 3-kinase-dependent signaling pathway.

Expression of deafness protein Tmie in postnatal developmental stages of C57BL/6J mice

Loss-of function mutations in the transmembrane inner ear expressed (Tmie/TMIE) gene have been shown to cause deafness in mice and humans (DFNB6). Previous studies report that the circling mouse can be an animal model for DFNB6. However, the expression pattern of Tmie protein in postnatal developmental stages has not been clearly revealed. In this study we tried to investigate the expression of Tmie protein in the liver, spleen, kidney, and lung, as well as in the cochlea. We examined various tissue samples from five different age groups of C57BL/6J animals. Using western blotting analysis, the expression of Tmie protein in these organs has been identified. The results show that Tmie protein expression in the cochlea has been increased in postnatal developmental stages, indicating that Tmie plays an important role in not only the development and also in the function of the cochlea. The expression pattern of Tmie in adult mouse organs such as the liver, spleen, kidney, and spleen significantly vary in adult rats. The order of Tmie expression level in mice (63 days after birth) was spleen, liver, lung, cochlea, and kidney, whereas in the adult rat it was liver, cochlea, lung, spleen, and kidney.

Over-expression of myosin7A in cochlear hair cells of circling mice

Circling mouse (C57BL/6J-cir/cir) deleted the transmembrane inner ear (Tmie) gene is an animal model for human non-syndromic recessive deafness, DFNB6. In circling mouse, hair cells in the cochlea have degenerated and hair bundles have become irregularity as time goes on. Tmie protein carries out a function of the mechanoelectrical transduction channel in cochlear hair cells. Myosin7a (MYO7A) protein has key roles in development of the cochlear hair bundles as well as in the function of cochlear hair cells. To find whether Tmie protein interacts with MYO7A proteins in the cochlea postnatal developmental stage, we investigated expression of the MYO7A proteins in the cochlear hair cells of circling mice by western blot analysis and whole mount immunofluorescence at postnatal day 5 (P5). The expression of MYO7A showed statistically significant increase in the cochlea of C57BL/6J-+/cir and C57BL/6J-cir/cir mice than that of C57BL/6J-+/+ mice. The MYO7A intensity of the cochlear hair cells also increased in C57BL/6J-+/cir and C57BL/6J-cir/cir mice compared with those of C57BL/6J-+/+ mice. Taken together, the results indicate that Tmie protein may have an important role with MYO7A protein in the development and maintenance of the stereociliary bundles during postnatal developmental stage of the cochlea.