• Journal of Ginseng Research
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• v.41 no.3
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• pp.386-391
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• 2017

Background: Korean Red Ginseng (KRG) is an ethnopharmacological plant that is traditionally used to improve the body's immune functions and ameliorate the symptoms of various diseases. However, the antitumorigenic effects of KRG and its underlying molecular and cellular mechanisms are not fully understood in terms of its individual components. In this study, in vitro and in vivo antitumorigenic activities of KRG were explored in water extract (WE), saponin fraction (SF), and nonsaponin fraction (NSF). Methods: In vitro antitumorigenic activities of WE, SF, and NSF of KRG were investigated in the C6 glioma cell line using cytotoxicity, migration, and proliferation assays. The underlying molecular mechanisms of KRG fractions were determined by examining the signaling cascades of apoptotic cell death by semiquantitative reverse transcriptase polymerase chain reaction and Western blot analysis. The in vivo antitumorigenic activities of WE, SF, and NSF were investigated in a xenograft mouse model. Results: SF induced apoptotic death of C6 glioma cells and suppressed migration and proliferation of C6 glioma cells, whereas WE and NSF neither induced apoptosis nor suppressed migration of C6 glioma cells. SF downregulated the expression of the anti-apoptotic gene B-cell lymphoma-2 (Bcl-2) and upregulated the expression of the pro-apoptotic gene Bcl-2-associated X protein (BAX) in C6 glioma cells but had no effect on the expression of the p53 tumor-suppressor gene. Moreover, SF treatment resulted in activation of caspase-3 as evidenced by increased levels of cleaved caspase-3. Finally, WE, SF, and NSF exhibited in vivo antitumorigenic activities in the xenograft mouse model by suppressing the growth of grafted CT-26 carcinoma cells without decreasing the animal body weight. Conclusion: These results suggest that WE, SF, and NSF of KRG are able to suppress tumor growth via different molecular and cellular mechanisms, including induction of apoptosis and activation of immune cells.

• 한국유가공학회:학술대회논문집
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• 1996.11a
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• pp.18-29
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• 1996

1. CWPC중의 새로운 생리활성물질의 검색 Mouse 임파세포의 증식효과를 지표로 하는 면역기능을 검토하여 CWPC중의 면역 부활작용을 갖는 새로운 성분의 검색을 실시하였다. CWPC를 여러 가지 분획법으로 분획하여 mouse 임파세포의 증식효과를 지표로 면역 활성성분을 검색하였다. 그 결과 gel filtration, 음이온교환법을 사용하여 분획한 당을 다량 포함한 부분에 강한 면역부활담당세포에 대하여 증식활성을 나타내는 물질을 발견하였다. 이 물질은 SDS-PAGE상에서 분자량이 약 16kDa에 위치하여 Ca, P 및 당쇄를 포함한 물질이며, 이것을 GPP로 하였다. GPP에는 우유케이신의 trypsin분해물이며 Ca와 무기인을 풍부하게 포함하는 ${\beta}$-CPP와 유사한 phosphoserin 영역을 갖는 성분과 갖지 않는 성분의 2종류가 존재하며, 각각의 면역 부활활성이 인정되었다. 각 성분의 아미노산 분석, 당 분석의 결과에서 지금까지 보고된 우유중의 면역 담당세포에 대한 증식활성을 갖는 물질과는 상이한 성분인 것으로 밝혀졌다. 더욱이 이 활성물질 (GPP)은 PP cell에서도 동등한 활성이 있는 것으로 판단되었다. 이러한 결과를 종합하여 보면 CWPC중에서 지금까지 알려지지 않았던 새로운 면역 부활물질이 존재하며, 그 성분에는 CPP와 유사한 phophoserine 영역이 존재하는 성분이 포함되어 있고, N-글리코실 결합의 당쇄가 존재하는 것으로 시사되었다. 이 성분은 전신면역의 지표인 비장세포에 대해서만이 아니고, 장관면역계에 중요한 역할을 담당하는 PP cell에서도 활성이 있는 것으로 보아 전신 및 국부적인 면역기능의 부활성분으로서 응용의 가능성이 시사되었다. 2. GPP의 면역담당세포에 대한 증식활성의 메카니즘의 검토 CWPC중의 GPP의 면역담당세포증식활성의 메카니즘을 해명하기 위해 먼저 이 성분중의 어느 부분이 활성에 관여하는지를 pronase 분해 및 phophoserine 영역을 인식하는 항체를 사용하여 검토하였다. 그 결과 pronase 분해처리에서도 활성의 감소를 나타내지 않았으므로 이러한 활성에는 당이 필수 불가결하다는 점이 시사되었다. 또한 phosphoserine 영역을 인식하는 항체에 의해서도 활성은 감소하지 않는 것으로 보아 phosphoserine 영역이 세포증식활성에 관여하지 않는 것으로 판단되었다. 또한 분획한 면역담당세포에 대한 증식활성을 측정하는 것으로 이 성분의 표적면역담당세포를 동정하여, B세포에 대해서만 특이적으로 증식활성을 나타내는 것으로 밝혀졌다.

• Korean Journal of Acupuncture
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• v.24 no.4
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• pp.111-129
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• 2007

Objectives & Methods : The purpose of this study is to observe the effects of Coicis Semen Herbal-acupuncture solution (CS-HAS) at the Joksamni (ST36) on the collagen-induced arthritis in the DBA/1J mouse. The author performed several experiments to analyze the effects of CS-HAS on arthritis; change of the weight; the spleen size and adhesion rate; serum cytokine levels; serum antibody levels; changes of immunocyte counts; the histological changes of joint. Results : In the Coicis Semen Herbal-acupuncture (CS-HA), arthritis index, the incidence of arthritis, and the degree of joint edema were decreased. In CS-HA, there was no weight loss. The size of the spleen, adhesion rate, and the edema and transformation of joint were lowered. In CS-HA, the levels of IL-1${\beta}$, IL-6, TNF-${\alpha}$, IFN-${\gamma}$, IgG, IgM, and anti-collagen II in serum and the levels of IFN-${\gamma}$, IL-4, IL-10 in spleen were significantly decreased. In CS-HA, the expression ratios of $CD45^+$ to $CD3e^+$ and $CD8^+$ to $CD4^+$ were decreased. Also, the overall $CD4^+/CD8^+$ cell ratio was lowered in spleen. Ratios of the $CD4^+/CD25^+$, $CD45^+/CD69^+$ cells were decreased in lymph nodes. In addition, ratios of the $CD3^+/CD69^+$, $CD11b^+/Gr-1^+$ cells were also decreased in synovium. In the histological study, the cartilage destruction and synovial cell proliferation, and collagen fiber destruction were decreased with CS-HA treated group. Conclusions : From the results mentioned above, it is suggested that CS-HA at the ST36 has several significant effects on the collagen-induced arthritis.

• Clinical and Experimental Reproductive Medicine
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• v.16 no.1
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• pp.9-14
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• 1989

For the cryopreservation of human embryos this study was accomplished as a preliminary experiment. The purpose of this study is to obtain optimal cryoprotectant, addition and dilution method of cryoprotectant and cooling rate for raising survival of frozon and thawed 2-cell mouse embryos. Seeding was done at $-7^{\circ}C$ and the straw contained embryos was plunged at $-30^{\circ}C$ when the slow cooling was ended. Embryos those developed normally to blastocyst after in vitro culture for over 96 hours were regarded as survival ones. The survival was the rate of number of survival embryos against the recovered embryos. The results are followed : 1. The survivals were 6.3, 71.2 and 67.4% respectively, when Glycerol, DMSO and 1,2-Propanediol were used as cryoprotectant. 2. When sucrose was added in freezing solution, the survival was 69.0%. That was higher than the survival of embryos frozen without sucrose in freezing solution. The difference was not significant. 3. Addition and dilution of cryoprotectant by 4 stepwise raised the survival than by direct, but that was not significant. 4. When embryos were frozen by -0.3, -0.5 and $-1^{\circ}C/min$ before plunged into $LN_2$, the survivals were 67.9, 78.0 and 37.0% respectively. The differnce was significant.

• BMB Reports
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• v.39 no.6
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• pp.686-695
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• 2006

Interleukin-2 enhancer binding factor 2 (ILF2) was reported to regulate transcription of interleukin-2 (IL-2), a central cytokine in the regulation of T-cell responses. This property of ILF2 was well characterized in human and mammals, but little is known in bony fish. In this paper, an ILF2 homologue was cloned and well characterized from Tetraodon nigrovirid is for the further investigation of the function of ILF2 in bony fish. The full-length Tetraodon ILF2 cDNA was 1380 bp in size and contained an open reading frame (ORF) of 1164 bp that translates into a 387 amino-acid peptide with a molecular weight of 42.9 kDa, a 5' untranslated region (UTR) of 57 bp, and a 3' UTR of 159 bp containing a poly A tail. The deduced peptide of Tetraodon ILF2 shared an overall identity of 58%~93% with other known ILF2 sequences, and contained two N-glycosylation sites, two N-myristoylation sites, one RGD cell attachment sequence, six protein kinase C phosphorylation sites, one amino-terminal RGG-rich single-stranded RNA-binding domain, and a DZF zinc-finger nucleic acid binding domain, most of which were highly conserved through species compared. Constitutive expression of Tetraodon ILF2 was observed in all tissues examined, including gill, gut, head kidney, spleen, liver, brain and heart. The highest expression was detected in heart, followed by liver, head kidney and brain. Stimulation with LPS did not significantly alter the expression of Tetraodon ILF2. Gene organization analysis showed that the Tetraodon ILF2 gene have fifteen exons, one more than other known ILF2 genes in human and mouse. Genes up- and down-stream from the Tetraodon ILF2 were Rpa12, Peroxin-11b, Smad4, Snapap and Txnip homologue, which were different from that in human and mouse.

• Journal of Haehwa Medicine
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• v.17 no.2
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• pp.167-183
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• 2008

KCSYJT medicines controlled $CD4^+/IFN-\gamma$, and $CD4^+/CD25^+/foxp3^+$ revelation that an experiment that motive allergy immune reponse because an in vitro experiment stimulates T cells of a NC/Nga mouse same time by anti-CD40/rmIL-4, and interleukin-$1{\beta}$, IL-6, TNF-$\alpha$, and TGF-$\beta$ mRNA outturn that bear in T and B cells decreased remarkably by KCSYJT medicines. Intracellular staining of splenocytes anti-CD40/rmIL-4 plus rmIL-4 stimulated as described in a, assessed after 24 h, KCSYJT exerts a mainly immunosuppressive effect that acts at least partially through suppression of the transcription factor GATA3 expression in $CD4^+$ T cells. We found that skin lesions, which were clinically and histologically very similar to human AD, mite antigen-induced dermatitis on the face, neck, ears and dorsal skin of inbred NC/Nga mice. Result that Th1 cell and Th2 cell observe to be shifted by cytokine expression with GATA3 regulation by KCSYJT medicines could know that KCSYJT medicines can use usefully in allergy autoimmnune diease.

• Archives of Pharmacal Research
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• v.19 no.3
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• pp.235-239
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• 1996

Six, heretofore undescribed, $5^I-Methyl-5^I-(5-Substituted uracil-1-ylmethyl)-2^I-oxo-3^I-methylenetetrahydrofurans(F, Cl, Br, l, CH_3, H)(6a-f)$were synthesized and evaluated against three cell lines (FM-3A, P-388 and U-937). For the preparation of .alpha.-methylene-.gamma.-butyrolactone bearing 5-substituted uracils (6a-f), the effcient Reformatsky type reaction was employed which involves the treatment of ethyl .alpha.(bromomethyl) acrylate and zinc with the respective 5-substituted uracil-1-ylacetones (5a-f). The acetone derivatives (5a-f) were directly obtained by the respective alkylation reaction of 5-substituted uracils with chloroacetone in the presence of $K_{2}$$CO_{3}$(or NaH). These lactone compounds 6a-f exhibited moderate to significant activity in all of the three cell lines, and 6b, 6c and 6e showed significant antitumor activities (inhibitory concentrations ($IC_{50}$) ranged from 1.3-3.8 .mu.g/ml.

• Journal of Animal Reproduction and Biotechnology
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• v.34 no.1
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• pp.20-29
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• 2019

Soy isoflavones have been reported to possess many physiological activities such as antioxidant activity and inhibition of cancer cell proliferation. This study investigated the photoprotective effects of soybean extract in human fibroblast cell line and hairless mice model. Human fibroblast was treated with soybean extract before and after ultraviolet B (UVB; 290-302 nm) irradiation. In the soybean extract treated group, the cells showed better resistance to ultraviolet (UV) than control group. The amount of type I collagen recovered from the soybean treated group was higher than the vehicle group exposed to UV-induced damage. Moreover, increased expression of metalloproteinases-1 as a result of UV irradiation was suppressed by the soybean extract. Female mice were orally administered soybean extract and irradiated with UVB light for 8 weeks. The effects of the soybean extract on the skin appearance, collagen deposition and epidermal thickness in the UV-damaged mouse skin were analyzed using histopathological methods. In soybean extract treated group, the skin had a better morphology than that of the control group. Furthermore, the amount of type I collagen was increased and overexpression of MMP-1 was reduced in the soybean extract group compared to vehicle group. Additionally, up-regulation of pro-inflammatory cytokines induced by UV irradiation was suppressed by dietary soybean extract treatment. It appears that soybean extract had a photoprotective effect, including anti-aging and anti-inflammatory effect, from UV-induced damage in not only human fibroblast, but also hairless mice. We confirmed that these effects were possibly due to promotion of collagen synthesis and inhibition of MMP-1 expression.

• Journal of Periodontal and Implant Science
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• v.32 no.4
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• pp.733-744
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• 2002

The fibroblasts are the principal cells in the periodontal ligament of peridontium. As the periodontal ligament fibroblasts (PDLF) show similar phenotype with osteoblasts, the PDLF are thought to play an important role in alveolar bone remodeling. Cell-to-cell contacted signaling is crucial for osteoclast formation. Recently it has been reported that PDLJ enhance the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The aims of this study were to $clarify\;^{1)}$ the mechanism of PDLF-induced osteoclastogenesis $and\;^{2)}$ whether we can use preosteoclast cell line instead of primary hematopoietic preosteoclast cells for studying the mechanism of PDLF-induced osteoclastogenesis. Osteoclastic differentiation of mouse macrophage cell line RAW264.7 was compared with that of mouse bone marrow-derived M-CSF dependent cell (MDBM), a well-known hematopoietic preosteoclast model, by examining, 1) osteoclast-specific gene expression such as calcitonin receptor, M-CSF receptor (c-fms), cathepsin K, receptoractivator nuclear factor kappa B (RANK) ,2) generation of TRAP(+) multinucleated cells (MNCs), and 3) generation of resorption pit on the $OAAS^{TM}$ plate. RAW264.7 cultured in the medium containing of soluble osteoclast differentiation Factor (sODF) showed similar phenotype with MDBM-derived osteoclasts, those are mRNA expression pattern of osteoclast-specific genes, TRAP(+) MNCs generation, and bone resorbing abivity. Formation of resorption pits by osteoclastic MNCs differentiated from sODF-treated RAW264.7, was completely blocked by the addition of osteoprotegerin (OPG), a soluble decoy receptor for ODF, to the sODF-containing culture me야um. The effects of PDLF on differentiation of RAW264.7 into　the TRAP(+) multinucleated osteoclast-like cells were examined using coculture system. PDLF were fxed with paraformaldehyde, followed by coculture with RAW264.7, which induced formation of TRAP(+) MNCs in the absence of additional treatment of sODF. When compared with untreated and fixed PDLF (fPDLF), IL-1 ${\beta}$-treated, or lipopolysaccha-ride-treated and then fixed PDLF showed two-folld increase in the supporting activity of osteoclastogenesis from RAW264.7 coculture system. There were no TRAP(+) MNCs formation in coculture system of RAW264.7 with PDLF of no fixation. These findigs suggested that we can replace the primary hematopoietic preosteoclasts for RAW264. 7 cell line for studying the mechanism of PDLF-induced osteoclastogenesis, and we hypothesize that PDLF control osteoclastogenesis through ODF expression which might be enhanced by inflammatory signals.

• Journal of Society of Preventive Korean Medicine
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• v.27 no.2
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• pp.23-33
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• 2023

Objectives : Sojadodamgangki-tang and its main components are traditional korean medicinal methods for treatment of cough, sputum and dyspnea. Using a respiratory inflammatory model, we intend to reveal the anti-inflammatory effect and its immune mechanism of Sojadodamgangki-tang. Methods : We used a papain-induced respiratory inflammatory mouse model. 8-week-old female BALB/C mice were divided into 3 groups as follows: the following groups: saline control group, papain treated group (vehicle), papain and Sojadodamgangki-tang(200 mg/kg) treated group (n=4). To evaluate the anti-inflammatory effect of Sojadodamgangki-tang extracts, inflammatory cell infiltration was measured in bronchoalveolar lavage fluid (BALF) and nasal lavage fluid (NALF). In addition, the effects of Sojadodamgangki-tang extracts on Th2 cell population in lung were determined by using flow cytometry. Results : Sojadodamgangki-tang extracts administration reduced inflammatory cell infiltration in BALF and NALF, especially of eosinophils. Furthermore, total immunogloblin (Ig)-E levels was reduced in BALF and serum by drug administration. Interestingly, Sojadodamgangki-tang extracts treatment also decreased the Th2 cell (CD4⁺GATA3⁺) population in lung. Conclusions : Our findings indicate Sojadodamgangki-tang extracts have anti-inflammatory effects by mediating Th2 cell and B cell activation.