• Journal of Nutrition and Health
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• v.48 no.2
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• pp.157-166
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• 2015

Purpose: The effects of water extract and distillate from the mixture of black goat meat and medicinal herb on MG-63 osteoblast proliferation and mouse bone marrow derived osteoclast formation were investigated. Methods: Proximate composition, volatile basic nitrogen (VBN), mineral content, free amino acid composition and free fatty acid composition in black goat meat were determined. Water extract and distillate were prepared with three groups; goat meat only (BG-E, BG-D), six herbs added group (BG-E6, BG-D6), and eight herbs added group (BG-E8, BG-D8). Osteoblast proliferation, mineralization and calcium uptake activity of MG-63 cells were measured and tartrate resistant acid phosphatase activity of osteoclasts was analyzed. Results: Black goat meat had remarkably low fat and high level of calcium. Glutamic acid was the most abundant amino acid. Herbs added extract groups (BG-E6 and BG-E8) showed increased MG-63 cell proliferation in a concentration dependent manner, while all the distillates did not show the effect. All extracts and distillates showed significantly increased osteoblast mineralization depending on the concentration. In particular, herb added extract, BG-E6, increased 170.3% of control and the distillate of BG-D and BG-D6 increased up to 168.5% and 159.8%, respectively. Calcium uptake activities of all water extracts showed remarkable increase of BG-E6 and BG-E8 up to 615.5% and 628.1% of control, respectively. Ditillates had no effect except BG-D6. All water extracts significantly reduced the activity of tartrate-resistant acid phosphatase (TRAP) in osteoclasts derived from mouse bone marrow. Conclusion: Combination of black goat meat and medicinal herb increased the MG-63 cell proliferation and effectively inhibited osteoclast differentiation in both water extracts and distillate of them, which implies that they could be used as potent functional food materials for bone health.

• Molecules and Cells
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• v.39 no.12
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• pp.855-861
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• 2016

Ginsenosides, which are the active materials of ginseng, have biological functions that include anti-osteoporotic effects. Aqueous ginseng extract inhibits osteoclast differentiation induced by receptor activator of NF-${\kappa}B$ ligand (RANKL). Aqueous ginseng extract produces chromatography peaks characteristic of ginsenosides. Among these peaks, ginsenoside Re is a major component. However, the preventive effects of ginsenoside Re against osteoclast differentiation are not known. We studied the effect of ginsenoside Re on osteoclast differentiation, RANKL-induced tartrate-resistant acid phosphatase (TRAP) activity, and formation of multinucleated osteoclasts in vitro. Ginsenoside Re hampered osteoclast differentiation in a dose-dependent manner. In an in vivo zebrafish model, aqueous ginseng extract and ginsenoside Re had anti-osteoclastogenesis effects. These findings suggest that both aqueous ginseng extract and ginsenoside Re prevent bone resorption by inhibiting osteoclast differentiation. Ginsenoside Re could be important for promoting bone health.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.9
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• pp.1476-1485
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• 2004

Of solvent-extracts prepared from the 90 kinds of Korean traditional tea and rice gruel plants, cold-water extract from peels of Citrus unshiu (CUI-0) showed the most potent intestinal immune system modulating activity through Peyer’s patch whereas other extracts did not have the activity except for cold-water extracts of Laminaria japonica, Polygonatum japonicum, Poncirus trifoliata, and hot-water extracts of Gardenia jasminoides, Lycium chinense having intermediate activity. CUI-0 was further fractionated into MeOH-soluble fraction (CUI-1), MeOH insoluble and EtOH-soluble fraction (CUI-2), and crude polysaccharide fraction (CUI-3). Among these fractions, CUI-3 showed the most potent stimulating activity for the proliferation of bone marrow cells mediated by Peyer’s patch cells, and contained arabinose, galacturonic acid, galactose, glucose, glucuronic acid and rhamnose (molar ratio; 1.00:0.53:0.45:0.28:0.28:0.19) as the major sugars, and a small quantity of protein (9.4%). In treatments of CUI-3 with pronase and periodate (NaIO₄), the intestinal immune system modulating activity of CUI-3 was significantly reduced, and the activity of CUI-3 was affected by periodate oxidation particularly. The potently active carbohydrate-rich fraction, CUI-3IIb-3-2 was further purified by anion-exchange chromatography on DEAE-Sepharose FF, Sepharose CL-6B and Sephacryl S-200. CUI-3IIb-3-2 was eluted as a single peak on HPLC and its molecular weight was estimated to be 18,000 Da. CUI-3IIb-3-2 was consisted mainly of arabinose, galactose, rhamnose, galacturonic acid and glucuronic acid (molar ratio;1.00:0.54:0.28:1.45:0.63) in addition to a small amount of proteins (3.2%). In addition, CUI-3IIb-3-2 showed the activity only through Peyer’s patch cells, but this fraction did not directly stimulate proliferation of bone marrow cells. It may be concluded that intestinal immune system modulating activity of peels from C. unshiu is caused by pectic polysaccharides having a polygalacturonan moiety with neutral sugars such as arabinose and galactose.

• Biomolecules & Therapeutics
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• v.29 no.2
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• pp.166-174
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• 2021

Multiple myeloma is a malignant cancer of plasma cells. Despite recent progress with immunomodulatory drugs and proteasome inhibitors, it remains an incurable disease that requires other strategies to overcome its recurrence and non-response. Based on the high expression levels of programmed death-ligand 1 (PD-L1) in human multiple myeloma isolated from bone marrow and the murine myeloma cell lines, NS-1 and MOPC-315, we propose PD-L1 molecule as a target of anti-multiple myeloma therapy. We developed a novel anti-PD-L1 antibody containing a murine immunoglobulin G subclass 2a (IgG2a) fragment crystallizable (Fc) domain that can induce antibody-dependent cellular cytotoxicity. The newly developed anti-PD-L1 antibody showed significant antitumor effects against multiple myeloma in mice subcutaneously, intraperitoneally, or intravenously inoculated with NS-1 and MOPC-315 cells. The anti-PD-L1 effects on multiple myeloma may be related to a decrease in the immunosuppressive myeloid-derived suppressor cells (MDSCs), but there were no changes in the splenic MDSCs after combined treatment with lenalidomide and the anti-PD-L1 antibody. Interestingly, the newly developed anti-PD-L1 antibody can induce antibody-dependent cellular cytotoxicity in the myeloma cells, which differs from the existing anti-PD-L1 antibodies. Collectively, we have developed a new anti-PD-L1 antibody that binds to mouse and human PD-L1 and demonstrated the antitumor effects of the antibody in several syngeneic murine myeloma models. Thus, PD-L1 is a promising target to treat multiple myeloma, and the novel anti-PD-L1 antibody may be an effective anti-myeloma drug via antibody-dependent cellular cytotoxicity effects.

• KSBB Journal
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• v.22 no.1
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• pp.22-29
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• 2007

The water-soluble materials extracted from fruit bodies and mycelium of H. erinaceum were prepared. In-vitro anticancer activities on cancer cells and In-vivo proliferation effect on mouse peritoneal exudate cell and spleen cell of samples were investigated. Also, nitric oxide (NO) generation of peritoneal exudate cell, IL-2 production capacity of spleen cells and phagocytic activity of peritoneal macrophages were examined. The water extracts of H. erinaceum suppressed the proliferation of cancer cell (HeLa, Raw264.7, Jurkat, KATO3, EL4, LyD9) with concentration-dependent. The water extract from fruit body showed better suppression effect than that from mycelium in most of cancer cells used. The anticancer effect of water extract of fruits body in the range of 0.01 and 10 mg/ml for Raw 264.7 and EL4 cell lines were the same as the Taxol with one thousandth equivalent of fruit body concentration. Water extracts of fruit body and liquid-cultured products of H. erinaceum induced nitric oxide (NO) generation of peritoneal exudate cell and increased NO generation by stimulus of lipopolysaccharide. Water extracts alone did not induce the proliferation and IL-2 production capacity of spleen cells. However, spleen's proliferation and IL-2 production were induced significantly by the addition of lipopolysaccharide and Con A (concanavalin A) or Con A alone, and the effectiveness of mycelium extract with water were more active than those from fruit body.

• IMMUNE NETWORK
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• v.11 no.1
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• pp.79-94
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• 2011

Background: Dendritic cell (DC)-based vaccines are currently being evaluated as a novel strategy for tumor vaccination and immunotherapy. However, inducing long-term regression in established tumor-implanted mice is difficult. Here, we show that deoxypohophyllotoxin (DPT) induces maturation and activation of bone marrow-derived DCs via Toll-like receptor (TLR) 4 activation of MAPK and NF-${\kappa}B$. Methods: The phenotypic and functional maturation of DPT-treated DCs was assessed by flow cytometric analysis and cytokine production, respectively. DPT-treated DCs was also used for mixed leukocyte reaction to evaluate T cell-priming capacity and for tumor regression against melanoma. Results: DPT promoted the activation of $CD8^+$ T cells and the Th1 immune response by inducing IL-12 production in DCs. In a B16F10 melanoma-implanted mouse model, we demonstrated that DPT-treated DCs (DPT-DCs) enhance immune priming and regression of an established tumor in vivo. Furthermore, migration of DPT-DCs to the draining lymph nodes was induced via CCR7 upregulation. Mice that received DPT-DCs displayed enhanced antitumor therapeutic efficacy, which was associated with increased IFN-${\gamma}$ production and induction of cytotoxic T lymphocyte activity. Conclusion: These findings strongly suggest that the adjuvant effect of DPT in DC vaccination is associated with the polarization of T effector cells toward a Th1 phenotype and provides a potential therapeutic antitumor immunity.

• Molecules and Cells
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• v.40 no.2
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• pp.123-132
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• 2017

Insulin signaling is coordinated by insulin receptor substrates (IRSs). Many insulin responses, especially for blood glucose metabolism, are mediated primarily through Irs-1 and Irs-2. Irs-1 knockout mice show growth retardation and insulin signaling defects, which can be compensated by other IRSs in vivo; however, the underlying mechanism is not clear. Here, we presented an Irs-1 truncated mutated mouse ($Irs-1^{-/-}$) with growth retardation and subcutaneous adipocyte atrophy. $Irs-1^{-/-}$ mice exhibited mild insulin resistance, as demonstrated by the insulin tolerance test. Phosphatidylinositol 3-kinase (PI3K) activity and phosphorylated Protein Kinase B (PKB/AKT) expression were elevated in liver, skeletal muscle, and subcutaneous adipocytes in Irs-1 deficiency. In addition, the expression of IRS-2 and its phosphorylated version were clearly elevated in liver and skeletal muscle. With miRNA microarray analysis, we found miR-33 was down-regulated in bone marrow stromal cells (BMSCs) of $Irs-1^{-/-}$ mice, while its target gene Irs-2 was up-regulated in vitro studies. In addition, miR-33 was down-regulated in the presence of Irs-1 and which was up-regulated in fasting status. What's more, miR-33 restored its expression in re-feeding status. Meanwhile, miR-33 levels decreased and Irs-2 levels increased in liver, skeletal muscle, and subcutaneous adipocytes of $Irs-1^{-/-}$ mice. In primary cultured liver cells transfected with an miR-33 inhibitor, the expression of IRS-2, PI3K, and phosphorylated-AKT (p-AKT) increased while the opposite results were observed in the presence of an miR-33 mimic. Therefore, decreased miR-33 levels can up-regulate IRS-2 expression, which appears to compensate for the defects of the insulin signaling pathway in Irs-1 deficient mice.

• BMB Reports
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• v.56 no.6
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• pp.359-364
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• 2023

KAI1/CD82, a membrane tetraspanin protein, can prevent various cancers and retinal disorders through its anti-angiogenic and anti-metastatic capacity. However, little is known about its anti-inflammatory effect and molecular mechanism. Therefore, the present study aimed to inLPSvestigate effect of a recombinant protein of the large extracellular domain of human KAI1 (Gly 111-Leu 228, rhKAI1) on lipopolysaccharides (LPS)-stimulated RAW264.7 macrophage-like cells and mouse bone marrow-derived macrophages (BMDM) and to identify its underlying mechanism. Our data showed that rhKAI1 suppressed expression levels of classically macrophages (M1) phenotype-related surface markers F4/80+CD86+ in LPS-stimulated BMDM and RAW264.7 cells. In addition, LPS markedly increased mRNA expression and release levels of pro-inflammatory cytokines and mediators such as interleukin (IL)-1β, IL-6, tumor necrosis factor-α, cyclooxygenase-2, nitric oxide and prostaglandin E2, whereas these increases were substantially down-regulated by rhKAI1. Furthermore, LPS strongly increased expression of NF-κB p65 in the nuclei and phosphorylation of ERK, JNK, and p38 MAPK. However, nuclear translocation of NF-κB p65 and phosphorylation of JNK were greatly reversed in the presence of rhKAI1. Especially, rhKAI1 markedly suppressed expression of toll-like receptor (TLR4) and prevented binding of LPS with TLR4 through molecular docking predict analysis. Importantly, Glu 214 of rhKAI1 residue strongly interacted with Lys 360 of TLR4 residue, with a binding distance of 2.9 Å. Taken together, these findings suggest that rhKAI1 has an anti-inflammatory effect on LPS-polarized macrophages by interacting with TLR4 and down-regulating the JNK/NF-κB signaling pathway.

• The Korean Journal of Pesticide Science
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• v.8 no.1
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• pp.22-29
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• 2004

To evaluate mutagenicity of Phosphinotricin Acetyltransferase(PAT) which is expressed by the glufosinate-resistance gene pat, in vitro reverse mutation test using Salmonella typhimurium, chromosome aberration test using chinese hamster lung(CHL) cells and in vivo micronucleus test of mice were performed. In the reverse mutation, the PAT did not induce mutagenicity in Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 with and without metabolic activation at $5000{\mu}g/plate$. In the chromosome aberration test, the results showed no incidence of increased structural and numerical chromosome aberrations at any doses tested(100, 10, $1{\mu}g/mL$). In micronucleus test, the ratio of micronuclei was measured in polychromatic erythrocytes of bone marrow of male ICR mice intraperitoneally administrated with PAT(1250, 625, and 313 mg/kg), the results showed no incidence of increased micronucleated polychromatic erythrocytes (MNPCE). These results indicate that PAT might not have mutagenic potential in vitro and vivo systems.

• Journal of Life Science
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• v.25 no.2
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• pp.223-230
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• 2015

Regulator of calcineurin 1 (RCAN1) is an endogenous calcineurin inhibitor that plays an important role in the pathogenesis of diseases related to the calcineurin-NFATc1 signaling pathway. The RCAN1-4 isoform is subject to NFATc1-dependent regulation. During receptor activator of nuclear factor kappa-B ligand (RANKL)-stimulated osteoclastogenesis, the calcineurin-NFATc1 pathway is critical. Because there is little information available on the role of RCAN1 in osteoclast differentiation, this study investigated whether changes in RCAN1 expression are related to the calcineurin-NFATc1 pathway and osteoclast differentiation. Mouse bone marrow monocytes (BMMs) were treated with 50 ng/ml of RANKL and M-CSF. Expression levels of NFATc1, calcineurin, and RCAN1 isoforms were determined using RT-PCR and Western blotting. Osteoclast differentiation was examined using tartrate-resistent acid phosphatase (TRAP) staining. To evaluate the effect of RCAN1 overexpression on osteoclastogenesis, cells were transfected with a mouse RCAN1-4 cDNA plasmid. After RANKL stimulation of BMMs, expression of NFATc1 and RCAN1 was increased at the mRNA and protein level, while calcineurin expression was unchanged. When the RCAN1-4 gene construct was transfected, the expression of RCAN1 protein was not increased despite several-fold increases in RCAN1-4 mRNA expression. Regardless of RANKL stimulation, over-expression of RCAN1-4 tended to reduce NFATc1 expression and knock-down of RCAN1 increase it. While BMMs transfected with the RCAN1-4 vector were differentiated into distinct osteoclasts, their phenotypes did not vary from those of mock controls. These results suggest that RCAN1 has a limited effect on the calcineurin-NFATc1 pathway during RANKL-stimulated osteoclast differentiation.