• Journal of the Korean Society of Food Science and Nutrition
• /
• v.41 no.10
• /
• pp.1378-1387
• /
• 2012

To investigate the pharmacological activity of chaga mushroom (Inonotus obliquus) on extraction conditions, chaga was extracted using water (reflux at $50^{\circ}C$, decoction over $90^{\circ}C$, pressure at $121^{\circ}C$) or ethanol (reflux at 50, 70, or $90^{\circ}C$). When water extract was further fractionated into crude polysaccharide (IO-CP), yields of IO-CP (4.8~16.8%) were higher than those of ethanolic extracts (IO-E, 1.9~2.7%) at increased temperature. For antioxidant activity, crude polysaccharide (IO-CP-121) obtained by pressurized extraction showed the highest polyphenolic and flavonoid contents (35.10 mg TAE/g and 18.48 mg QE/g, respectively) as well as DPPH and ABTS free radical scavenging activities (26.08 and 27.99 mg AEAC/100 mg, respectively). Meanwhile, IO-CP-D (decoction) and IO-CP-50 (reflux) had more potent mitogenic effects (2.10- and 1.95-fold of saline control at 100 ${\mu}g/mL$) as well as intestinal immune system modulating activities (6.30- and 5.74-fold) compared to IO-CP-121, whereas ethanolic extracts showed no activity. Although no IO-CP showed cytotoxicity against RAW 264.7 cells at 0.1 mg/mL, IO-CP-121 significantly inhibited TNF-${\alpha}$ and NO production as pro-inflammatory factors in LPS-stimulated RAW 264.7 cells (29.2 and 63.5%, respectively). Ethanolic extracts also showed no cytotoxicity at 0.1 mg/mL, whereas inhibition of TNF-${\alpha}$ and NO production was significantly low compared to that of IO-CP-121. In addition, active IO-CP-D was further fractionated into an unadsorbed (IO-CP-I) and seven adsorbed fractions (IO-CP-II~VIII) by DEAE-Sepharose CL-6B column chromatography in order to isolate immunostimulating polysaccharide. IO-CP-II showed the most potent mitogenic effect and macrophage stimulating activity (4.51- and 1.64-fold, respectively). IO-CP-II mainly contained neutral sugars (61.86%) in addition to a small amount of uronic acid (2.96%), and component sugar analysis showed that IO-CP-II consisted mainly of Glc, Gal, and Man (molar ratio of 1.00:0.55:0.31). Therefore, extraction conditions affect the physiological activity of chaga, and immunostimulating polysaccharide fractionated from chaga by decoction is composed mainly of neutral sugars.

• Korean Journal of Veterinary Research
• /
• v.34 no.3
• /
• pp.457-463
• /
• 1994

Insulin-like growth factor-I(IGF-I) plays an important role in the regulation of mammalian and poultry growth. IGF-I has many actions in different tissues, which include metabolic, mitogenic, and differentiative actions. IGF-I induces insulin-like effects - such as increased cell glucose uptake and glycogen sysnthesis, however several physiological actions of IGF-I may not have been identified yet. In order to investigate the effect on growth in broiler chicken treated with exogenous insulin-like growth factor-I, 30 chickens were injected $50{\mu}g$ reconbinant human IGF- I (rhIGF- I ) per kg body weight as experimental group and 30 ckickens saline subcutanously as control, 3 times according to ages from 2 to 35 days. We established radioimmunoassay method by which we can measure chicken IGF- I (cIGF- I ) as in rhIGF- I assay. The results obtained were as follows; 1) The dilution curve showed in parallelism between rhIGF- I and cIGF- I in the Sep-pak $C_{18}$ cartridge plasma extracts. 2) The body weight of broiler chicken were significantly increased at 31 days($1,176.50{\pm}99.79g$) and 35 days($1,252.84{\pm}125.21g$) of age in treatment groups, compared with control group($1,011.88{\pm}40.22g,\;1,111.32{\pm}153.67g$). The liver and kidney weights on 35 days$(35.24{\pm}5.18g,\;11.05{\pm}1.47g)$ were significantly higher in rhIGF- I treated group than control group($30.95{\pm}4.04g,\;10.01{\pm}1.60g$) 3) The plasma concentration of IGF- l and total protein in rhIGF- I treated group were $58.17{\pm}1.69ng/ml$, $3.75{\pm}0.62g/dl$ respectively compared with control group $45.70{\pm}1.64ng/ml$, $2.32{\pm}0.53g/dl$. The results suggest that exogenous rhIGF- I increased total body weight, liver and kidney weights in broiler chicken, and it may increase IGF- I and total protein concentration in serum.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.34 no.5
• /
• pp.550-555
• /
• 2001

Insulin-like growth factor-I (IGF-I) is a mitogenic peptide with a molecular mass of 7 kDa. It is produced mainly in the liver and has important functions in the regulation of development and somatic growth. Moreover, Serum IGF-I concentration is regulated by the quantity and the nutritional quality of dietary protein. To determine the IGF-I level in Korean rockfish, Sabastes schlegeli, were fed four experiment diets that contained different protein quantities, namely $30\%,\;40\%,\;50\%\;and\;60\%$ for 70 days. Weight gain of the fish increased depending dietary protein quantity, Also, IGF-I concentrations increased according to dietary protein quantity, Feeding experiments were conducted to examine the effects of dietary protein sources on the serum IGF-I level in Korean rockfish, Fish meal (CO), soybean meal (SM), corn-gluten meal (CGM), meat meal (MM) and feather meal (FM) were used as variable protein sources of the formulated diet. IGF-I concentrations of the CO and MM groups ($277.7\pm23.2,\;291.5\pm41.2\;ng/mL$) were higher than those of the CGM and FM groups ($208.9\pm21.3,\;217.2\pm38.2\;ng/mL$). And IGFBP-3 levels by western blot analysis increased in good protein diets such as in the CO and MM groups. In conclusion, IGF-I may be a sensitive indicator the protein metabolism in fish as well as mammalian.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.18 no.4
• /
• pp.1021-1027
• /
• 2004

Panax ginseng has been used as a typical tonic medicine in Asian countries, such as Korea, China, and Japan. It has been reported that ginsenoside Rg1 in Panax ginseng increases the proportion of T helper cells in the whole T cells and promotes IL-2 gene expression in murine splenocytes. These studies imply that ginsenoside Rg1 increases the immune activity of CD4+ T cell, however the exact mechanism of ginsenoside Rg1 on helper T cell remains to be verified. The present study tried to elucidate the direct effect of Rg1 on helper T cell s activities and its Th1/Th2 lineage development. The results demonstrated that ginsenoside Rg1 had not mitogenic effects on the unstimulated CD4+ T cell, but augmented CD4+ T cell proliferation upon activating with anti-CD3/anti-CD28 antibodies in a dose dependent manner. Rg1 also enhanced the expression of cell surface protein CD69 on CD4+ T cell. In Th0 condition, ginsenoside Rg1 increases the expression of IL-2 mRNA, and enhances the expression of IL-4 mRNA on CD4+ T cells, suggesting Rg1 prefer to induce Th2 lineage development. In addition, ginsenoside Rg1 increases IL-4 secreting CD4+ T cell under Th2 skewed condition, while decreases IFN-γ secreting cell in Th1 polarizing condition. Thus, Rg1 enhances Th2 lineage development from naive CD4+ T cell both by increasing Th2 specific cytokine secretion and by repressing Th1 specific cytokine production. Therefore, these results suggest that ginsenoside Rg1 might be desirable agent for enhancing CD4+ T cell's activity, as well as the correction of Th1 dominant pathological disorders.

• Journal of Korean Neurosurgical Society
• /
• v.44 no.6
• /
• pp.375-381
• /
• 2008

Objective : The effects on neural proliferation and differentiation of neural stem cells (NSC) of basic fibroblast growth factor-2 (bFGF). insulin growth factor-I (IGF-I). brain-derived neurotrophic factor (BDNF). and nerve growth factor (NGF) were assessed. Also, following combinations of various factors were investigated : bFGF+IGF-I, bFGF+BDNF, bFGF+NGF, IGF-I+BDNF, IGF-I+NGF, and BDNF+NGF. Methods : Isolated NSC of Fisher 344 rats were cultured with individual growth factors, combinations of factors, and no growth factor (control) for 14 days. A proportion of neurons was analyzed using $\beta$-tubulin III and NeuN as neural markers. Results : Neural differentiations in the presence of individual growth factors for $\beta$-tubulin III-positive cells were : BDNF, 35.3%; IGF-I, 30.9%; bFGF, 18.1%; and NGF, 15.1%, and for NeuN-positive cells was : BDNF, 34.3%; bFGF, 32.2%; IGF-I, 26.6%; and NGF, 24.9%. However, neural differentiations in the absence of growth factor was only 2.6% for $\beta$-tubulin III and 3.1% for NeuN. For $\beta$-tubulin III-positive cells, neural differentiations were evident for the growth factor combinations as follows : bFGF+IGF-I, 73.1 %; bFGF+NGF, 65.4%; bFGF+BDNF, 58.7%; BDNF+IGF-I, 52.2%; NGF+IGF-I, 40.6%; and BDNF+NGF, 40.0%. For NeuN-positive cells : bFGF+IGF-I, 81.9%; bFGF+NGF, 63.5%; bFGF+BDNF, 62.8%; NGF+IGF-I, 62.3%; BDNF+NGF, 56.3%; and BDNF+IGF-I, 46.0%. Significant differences in neural differentiation were evident for single growth factor and combination of growth factors respectively (p<0.05). Conclusion : Combinations of growth factors have an additive effect on neural differentiation. The most prominent neural differentiation results from growth factor combinations involving bFGF and IGF-I. These findings suggest that the combination of a mitogenic action of bFGF and post-mitotic differentiation action of IGF-I synergistically affects neural proliferation and NSC differentiation.

• Journal of Periodontal and Implant Science
• /
• v.32 no.3
• /
• pp.457-473
• /
• 2002

The purposes of this study is to evaluate the combination effects of TGF-${\beta}_1$ and PDGF-BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/100% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Authors measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis according to the concentration of TGF-${\beta}_1$,(1,5ng/ml) and PDGF-BB (1,10 ng/ml) in combination. To explore further this delayed effect of TGF-${\beta}_1$, we preincubated human periodontal ligament cells with TGF-${\beta}_1$ for 4 or 24 hours before PDGF-BB stimulation. The results were as follows: The DNA synthetic activity was increased dose dependently by TGF-${\beta}_1$, PDGF-BB. The combination of TGF-${\beta}_1$ and PDGF-BB consistently enhanced the DNA synthetic activity to PDGF-BB alone. The ability of TGF-${\beta}_1$ to enhance DNA synthetic activity in PDGF-BB treated periodontal ligament cells was dose dependent. The maximum mitogenic effect was at the 5ng/ml of TGF-${\beta}_1$ and l0ng/ml of PDGF-BB. Preincubation of cell with TGF-${\beta}_1$ resulted in significantly greater response to PDGF-BB at all TGF-${\beta}_1$ concentration studied, and may be useful for clinical application in periodontal regenerative procedures. The total protein, collagen and noncollagen synthesis was increased dose pendently by TGF-${\beta}_1$, PDGF-BB. The % of collagen was slightly decreased according to the concentration of TGF-${\beta}_1$, PDGF-BB. The effect of TGF-${\beta}_1$, PDGF-BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. This study demonstrates that PDGF-BB is major mitogens for human periodontal ligament cells in vitro, and supports a role for TGF-${\beta}_1$ as a regulation of the mitogenic and total protein formation to PDGF-BB in these cells.

• Journal of Periodontal and Implant Science
• /
• v.24 no.2
• /
• pp.303-320
• /
• 1994

Current acceptable methods For promotin gperiodontal regeneration are base on removal of diseased soft tissue, root treatment, guided tissue regeneration, inteoduction of new graft materials and biological mediators. Platelet-derived growth factor(PDGF) is one of polypeptide growth factor. PDGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purposes of this study is to evaluate the effects of PDGF-AA, BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/10% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Author measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis and alkaline phosphatase activity according to the concentration of PDGF-AA and BB(0, 0.1, 1, 10, 100ng/ml). The results were as follows : The DNA synthetic activity was increased dose dependently by PDGF-AA and BB. The maximum mitogenic effect was at the 100ng/ml of PDGF-AA and 10ng/ml of PDGF-BB. The total protein, collagen and noncollagen systhesis was increased dose dependently by PDGF-AA and BB. The % of collagen was slightly decresed according to the concentration of PDGF-AA and BB. The effect of PDGF-AA and BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. The effect of PDGF-AA and BB on alkaline phosphatase activity did not show any significant, meanwhile the alkaline phosphatase activity of 14 days group showed significnat increase. In conclusion, PDGF-AA and BB may have important roles in stimulation of DNA synthesis in human periodontal ligament cells, which means an increase in collagen-synthesizing cells, and may be useful for clinical application in periodontal regenerative procedures.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.40 no.9
• /
• pp.1333-1339
• /
• 2011

After mulberry (Morus alba) leaves were fermented with Hericium erinaceum mycelium by solid-state culture to enhance physiological activity, fermented mulberry leaves (MA-HE) was extracted by hot-water (MA-HEHW) and ethanol (MA-HE-E). MA-HE-HW showed enhanced mitogenic and intestinal immune system modulating activities (1.41 and 1.52 fold of saline control, respectively) compared to hot-water extracts of non-fermented mulberry leaves (MA-HW) and H. erinaceum mycelium (HE-HW) at $100\;{\mu}g$/mL. Meanwhile, when we tested the inhibitory effects of extracts on nitric oxide (NO), tumor necrosis factor (TNF)-${\alpha}$, and interleukin (IL)-$1{\beta}$ and IL-6 production, MA-HE-E significantly inhibited these pro-inflammatory mediators in LPS-stimulated RAW 264.7 cells (45.1, 41.3, 70.2, and 55.7% inhibition of LPS control at $1,000\;{\mu}g$/mL). In addition, MA-HE-HW and MA-HE-E did not show any cytotoxicity on RAW 264.7 cells at $1,000\;{\mu}g$/mL whereas HE-E and MA-E indicated cytotoxicity (80.1 and 30.7% cell viability of saline control). These results suggest that mulberry leaves fermented with H. erinaceum by solid-state culture might have enhanced immunomodulatory and anti-inflammatory effects compared to non-fermented mulberry leaves, resulting in ingredients biotransformed for fermentation with H. erinaceum mycelium.

• Journal of Periodontal and Implant Science
• /
• v.26 no.2
• /
• pp.469-490
• /
• 1996

The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the effect on the cell activity than $TGF-{\beta}1$ only tested group and the tested group of minocycline inhibited cell activity. This results indicate that minocycline was effective on cell attachment in early stage, but it is harmful to cell activity, that inhibitory effect of minocycline was compensated with stimulatory effect of $TGF-{\beta}1$.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.38 no.9
• /
• pp.1145-1152
• /
• 2009

For the utilization of Korean ginseng (Panax ginseng C.A. Meyer) in the functional drink, we prepared the fermented Korean ginseng with mushroom mycelia (Ganoderma lucidum; WG-GL, Hericium erinaceum; WG-HE and Phellinus linteus; WG-PL) by solid culture. A proximate analysis showed that the fermented Korean ginseng contained significantly more crude fat (4.66$\sim$12.02%) than Korean ginseng (WG, 1.61%) whereas crude protein content of WG (13.64%) was higher value than those of the ferments (7.60$\sim$12.57%). When we also evaluated effects of the fermented Korean ginseng on the mitogenic activity, hot-water extract from WG-PL was significantly higher than those of WG or mycelia only fermentation (GL, HE and PL) as analyzed by IL-2 production (1.64-fold of the saline control) and proliferation of splenocytes (1.47-fold). In addition, the lysosomal phosphatase activity (WG-HE; 1.32-fold) and NO/TNF-$\alpha$ production (WG-HE; 2.27-fold of the saline control at 50 ${\mu}g$/mL, WG-PL; 3.56-fold, respectively) from macrophage in the presence of the fermented Korean ginseng were higher than those of WG or mycelia fermentation. These results indicate that hot-water extracts from the fermented Korean ginseng with mushroom mycelia by solid culture contain chemical ingredients different from the Korean ginseng, and that it might provide beneficial immunostimulating activity.