• 한국발생생물학회지:발생과생식
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• 제2권1호
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• pp.29-37
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• 1998

포유류 배아의 초기발생과정에서 수란관이 하는 역할을 알아보기 위하여 소의 수란관내액이 생쥐 2-세포기 배아의 체외발생에 미치는 영향을 조사하였다. 대조군의 경우 5%의 배아만이 체외배양과정 중 퇴화한데 비해 5% 혹은 그 이상의 소의 수란관내액 (bOF)이 함유된 배양액 내에서 배양된 배아는 48시간이 지났을 때 모두 퇴화하였다. bOF를 65 \circ C에서 30분간 가열한 후 배양액에 첨가한 결과 bOF의 독성은 변화가 없었으나 90 \circ C에서 30분간 가열하였을때에는 독성이 거의 사라져 95% 의 배아가 정상적으로 발생하였다. bOF를 chymotrypsin으로 1시간 혹은 24시간동안 처리한 후 배양액에 첨가한 경우에도 독성이 사라져 각각 95.5%의 배아가 상실배 혹은 포배로 발생을 진행하였다. 산화방지제인 10mM 농도의 glutathione (GSH)을 bOF와 함께 배양액에 첨가한 결과 마찬가지로 bOF의 독성이 사라져 91.0%의 배아가 상실배 이상으로 발생한 반면 산화 방지능력이 없는 산화형 GSH(GSSG)를 bOF와 함께 배양액에 처리한 경우에는 bOF의 독성은 전혀 제거되지 않았다. 또한 mercaptoethanol, dithiothreitol, cysteamine 등의 다른 산화방지제도 GSSG와 마찬가지로 bOF의 독성을 제거하지 못하였다. 이와 같은 실험 결과로 미루어 소의 수란관내액에는 체외에 노출될 경우 독성을 나타내는 단백질 성분으로 된 물질이 있으며 이 물질의 독성은 대기 중의 산소의 산화작용에 의해 활성화되는 것으로 여겨진다.

• Radiation Oncology Journal
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• 제25권3호
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• pp.185-191
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• 2007

목적 : 가시오갈피 뿌리 추출물(Extract of Eleuthrococcus senticosus Maxim root, ESMR)의 면역세포의 활성에 미치는 효과와 마우스 종양세포에서 세포독성 및 방사선 병용효과를 알기위해서 본 연구를 시행하였다. 대상 및 방법: 가시오갈피 뿌리(250 g)를 80% 메탄올로 추출한 다음, 감압농축 및 동결과정을 통해 분획을 얻었다. 세포 주는 섬유육종세포를 이용하였으며, 방사선조사는 저 에너지(250 KV)용 방사선 기기를 사용 하였다. 가시오갈피 추출물의 면역세포에 미치는 활성효과를 알아보기 위해서 마우스 비장 및 흉선세포의 생존율 측정과 비장 및 흉선림프구 아집단을 측정하였다. 세포독성 정도를 알아보기 위하여 clonogenic assay법을 이용하였다. 방사선조사 단독군의 경우 2, 4 및 6 Gy를 마우스 섬유육종세포(FSa II)에 조사하였으며, 가시오갈피와 방사선 병용군의 경우 0.2 mg/ml 농도의 가시오갈피를 방사선조사 1시간 전에 마우스 섬유육종세포에 접촉시켰다. 결 과: 가시오갈피 /뿌리 추출물 각각 10 ${\mu}g/ml$과 $100{\mu}g/ml$을 첨가한 마우스 비장 및 흉선세포 생존율은 다음과 같았다. 마우스 비장세포는 각각 $8.8{\pm}1.7%$와 $37.7{\pm}2.1%$로 증가하였으며, 흉선세포에서는 100 ${\mu}g/ml$에서 $16.7{\pm}1.6%$로 증가하였다. ESMR을 경구 투여한 마우스 비장 및 흉선림프구는 다음과 같았다. 마우스 비장세포의 B cell은 41.5 $({\pm}1.3)$에서 46.0 $({\pm}1.5)%$로, T cell은 16.9 $({\pm}0.9)$에서 22.3 $({\pm}1.2)%$로 증가하였으며, 특히 Helper T (Th) cell은 10.4 10.4 $({\pm}0.7)$에서 14.2 $({\pm}0.8)%$로, Cytotoxic T (Tc) cell은 3.6 $({\pm}0.4)$에서 5.6 $({\pm}0.3)%$로 증가하였다. 그러나 마우스 흉선세포에서 T림프구의 증가율은 거의 보이지 않았다. 마우스 섬유육종 세포에서 ESMR의 세포독성을 나타내는 생존분율은 0.2, 2.0 mg/m에서 각각 $0.36{\pm}0.02$, $0.05{\pm}0.02$으로 나타났다. 마우스 섬유육종 세포를 접종한 마우스에 ESMR 0.2 mg/ml 농도를 1시간 동안 접촉시킨 다음, 방사선조사량 2, 4 및 6 Gy를 조사한 후 얻은 생존분물은 0.39, 0.22 및 0.06이었으며, 방사선조사 단독군의 경우는 0.76, 0.47 및 0.37로 나타났다. 그리고 두 군간의 차이는 통계학적 의미를 나타냈다(p<0.05). 결 론: 가시오갈피 뿌리 추출물은 in vitro 실험에서 마우스 비장세포의 생존율을 증가시켰다. In vivo 실험에서는 마우스 비장 내 B세포 및 T세포를 증가시켰으나, 마우스 흉선세포에 대해서는 증가를 보이지 않았다. ESMR은 마우스 섬유육종 세포에 대해 강한 세포독성 효과를 나타냈다. FSa II를 접종한 마우스에서 방사선 단독 보다 ESMR을 병용한 경우 방사선에 의한 세포손상이 약 50% 이상 증가되었으며, 두 군간의 차이는 통계학적 의미를 나타냈다(p<0.05).

• Journal of Acupuncture Research
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• 제18권1호
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• pp.186-201
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• 2001

Purpose ; In order to study the effect of Houttuyniae herbal acupuncture on growth of melanoma B 16 and immunity in mice, the Control group with normal saline acupunchure after subcutaneous inoculation of B16BL6 cells, the Sample I with Houttuyniae herbal acupuncture manufactured by distilled water extraction method after subcutaneous inoculation of B16BL6 cells and the Sample II with Houttuyniae herbal acupuncture manufactured by alcohol extraction method after subcutaneous inoculation of B16BL6 cells were divided. Methods ; To evauluate the effect of Houttuynias herbal acupuncture on growth of melanoma B16 in mice, the weight of mouse melanoma, body weight, spleen index( $\sqrt(Weight of spleen/Body weight){\times}100$), lymphocytes numbers in mouse peripheral blood and spleen tissue, the percentage of CD4+ T cells, CD8+ and the ratio of CD4+/CD8+ in mouse peripheral blood and spleen tissue are measured. Results ; In study of the weight of mouse melanoma, Houttuyniae herbal acupuncture groups(Sample I, Sample II) showed statistically significant inhibitory effect. And in study of effect of reduction of change in body weight of mouse, Sample I showed statistically significant inhibitory effect, too. In study of reduction of spleen index, lymphocytes numbers in mouse spleen tissue, and the percentage of CD4+ and the ratio of CD4+/CD8+ T cells in mouse peripheral blood, Houttuyniae herbal acupuncture groups(Sample I, Sample II ) showed with statistically significant inhibitory effect. Also, Sample I showed inhibitory effect of reduction of the percentage of CD8+ T cells in mouse peripheral blood and spleen tissue and Sample II showed inhibitory effect of reduction of the percentage of CD4+ and the ratio of CD4+/CD8+ T cells in mouse spleen tissue with statistical significance. Conclusions ; The inhibitory effect of reduction of lymphocytes numbers in mouse peripheral blood and IL-2 productivity, Houttuyniae herbal acupuncture groups(Sample I, Sample II) didn't show significant effect.

• 대한한방소아과학회지
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• 제29권4호
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• pp.39-66
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• 2015

Objectives The purpose of this study is to investigate the effects of Sesamum indicum extracted (SEI) on atopic dermatitis in an in-vitro and in-vivo experiment using a MC/9 murine mast cells and a NC/Nga mouse. Methods In-vitro experiment, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF mRNA expression were evaluated by Real-time PCR, IL-13, MIP-$1{\alpha}$ production by ELISA and manifestations of NFAT-1, NFAT-2, c-jun, c-fos, NF-${\kappa}B$ p65 transcription factors by western blotting. In-vivo experiment, we measured WBC, Eosinophil, Neutrophil, and serum IL-5, IL-13 in NC/Nga atopic dermatitis mouse, IL-5, IL-13, IFN-${\gamma}$, IL-4 in the spleenocyte culture supernatant by ELISA, the absolute cell numbers of CD4+, CD8+, +Gr-1+CD11b, B220+CD23+ in the axillary lymph node (ALN), peripheral blood mononuclear cells (PBMCs) and dorsal skin tissue, IL-5, IL-13 by Real-time PCR, the distribution of tissue inflammation and cellular infiltration by H&E and toluidine blue. Results SEI decreased IL-4, IL-5, IL-6, IL-13, GM-CSF, TNF-${\alpha}$ mRNA expression, IL-13, MIP-$1{\alpha}$ production and the expression of transcription factors including NFAT-1, c-jun, NF-${\kappa}B$ p65 in MC/9 murine mast cells. SEI orally administration decreased cell number of WBC, Eosinophil, the level of serum IgE, total cell number of ALN and dorsal skin tissue, absolute cell number of CD4+, CD8+, B220+CD23+ in the ALN. SEI orally administration also increased absolute cell number of CD8+/CD3+ and decreased Gr-1+/CD11b+ in PBMCs, decreased CD4+ in dorsal skin tissue, inhibited IL-5, IL-13 mRNA expression. Infiltration levels of inflammatory immune cells, mast cells and thickness of epidermis decreased in dorsal skin tissue. Conclusions SEI can regulate allergic inflammatory response suppressed the gene expression and production of cytokines that mediate allergic reactions, and will be able to be effectively utilized in the treatment of atopic dermatitis future.

• International Journal of Industrial Entomology and Biomaterials
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• 제7권2호
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• pp.197-201
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• 2003

The recombinant plasmids harboring a heterologous gene coding mouse endostatin were transfected and expressed stably in Trichoplusia ni Tn 5B1-4 cells and Bombyx mori BmN cells, respectively. Recombinant endostatin expressed in the stably transformed Tn 5B1-4 and BmN cells was secreted into the medium. BmN cells are relatively lower in maximum cell growth and recombinant endostatin production than Tn 5B 1-4 cells. Recombinant endostatin was also purified to homogeneity using a simple one-step ${Ni^2+}$ affinity fractionation method. Purified recombinant endostatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition $({ED_50})$ for recombinant endostatin was approximately 0.35 ${\mu}g$/ml.

• 대한한방소아과학회지
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• 제30권4호
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• pp.29-59
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• 2016

Objectives PRAL (Prunus armeniaca Linne Var) has been known to suppress allergic reaction. However, the cellular target and its mechanism of action were unclear. This study was designed to investigate the effect of PRAL on RBL-2H3 mast cell, which is PMA-Ionomycin-induced activated in vitro and the effect of PRAL on the MNC/Nga mice that are DNCB-induced activated in vivo. Methods In this study, IL-4, IL-13 production were examined by ELISA analysis; IL-4, IL-13, IL-31, IL-31Ra, $TNF-{\alpha}$ and GM-CSF mRNA expression were examined by Real-time PCR; manifestations of AP-1 and MAPKs transcription factors were examined by western blotting in vitro. Then skin rashes have been evaluated and verified the distribution of mast cells by H&E and toluidine blue. Also, WBC, eosinophil and neutrophil, IgE level in serum, $IFN-{\gamma}$, IL-4, IL-5 in the splenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$, $CD3^+CD69^+$ in the Axillary Lymph Node (ALN), PBMCs and dorsal skin and IL-5, IL-13, IL-31, IL-31Ra in the dorsal skin by Real-time PCR were all evaluated from the NC/BNga mice. Results As a result of this study, the mRNA expression of IL-4, IL-13, IL-31, IL-31Ra and $TNF-{\alpha}$ and IL-4, IL-13 production, shown in ELISA analysis, were suppressed by PRAL. Results from the western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including AP-1 and p-JNK, p-ERK. Histological examination showed that infiltration levels of immune cells in the skin of the AD-induced NC/Nga mice were improved by PRAL orally adminstration. Orally- administered PRAL group also showred decreased level of IgE in the serum. This group has shown decreased the level of IL-4, IL-5, but shown elevated $IFN-{\gamma}$ level in the splenocyte culture supernatant. The same group also has shown decreased cell numbers of $CD4^+$, $CD8^+$, $CD3^+CD69^+$ in the ALN, and $CD4^+$, $Gr-1^+CD11b^+$ in the dorsal skin. PRAL oral adminstration increased cell numbers of $CD4^+$, but decreased cell numbers of $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$ in the PBMCs. Conclusions Obtained results suggest that PRAL can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis (AD) induced in the NC/Nga mice. This may play an important role in recovering AD symptoms and suppressing pruritus.

• 환경위생공학
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• 제20권3호
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• pp.10-16
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• 2005

Chestnut bark extract by methanol repressed the expression of tyrosinase gene of B16 mouse melanoma cell containing tyrosinase promoter. $10{\mu}g/ml{\ell}\;100{\mu}g/m{\ell}$, $1mg/m{\ell}$ of the extract repressed expression of tyrosinase gene about $38\%,\;47\%,\;and\;78\%$, respectively, compared with control. In the MTT assay, the same extract exhibited very low cytotoxicity at $1{\mu}g/m{\ell},\;10{\mu}g/m{\ell},\;100{\mu}g/m{\ell},\;and\;1mg/m{\ell}$, respectively. The fractions of Methylene chloride and ethyl acetate did not showed the repressive effect on the expression of tyrosinase gene, but the fraction of butyl alcohol repressed highly at $10{\mu}g/m{\ell}\;and\;100{\mu}g/m{\ell}$.

• IMMUNE NETWORK
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• 제12권3호
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• pp.84-88
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• 2012

B cell-activating factor belonging to the TNF family (BAFF) is primarily expressed by macrophages and stimulates B cell proliferation, differentiation, survival, and Ig production. In this study, we explored the effect of lactoferrin (LF) on BAFF expression by murine macrophages. We determined the level of BAFF expression at the transcriptional and protein levels using RT-PCR and ELISA, respectively. LF markedly enhanced BAFF expression in mouse macrophages at both the transcriptional and protein levels. Overexpression of Smad3/4 further increased LF-induced BAFF transcription while DN-Smad3 abolished the LF-induced BAFF expression. These results demonstrate that LF can enhance BAFF expression through Smad3/4 pathway.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.260.1-260.1
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• 2003

Magnoliae Flos(‘shin-j’). the flower buds of Magnolia kobus, is acrid to taste with a ‘warm’ property. It is a ‘wind-cold’ discutient and nasal decongestant and is principally used in the treatment of nasal congestion with headache, sinusitis and allergic rhinitis. By screening inhibitory activities on the melanin polymer biosysthesis in B-16 mouse melanoma cell lines, methylene chloride extract of Magnoliae Flos was found to have inhibitory effect on melanin polymer biosynthesis. (omitted)

• 대한한방내과학회지
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• 제21권1호
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• pp.156-161
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• 2000

The unripe fruit of Poncirus trifoliata Raf has been used for the treatment of allergic disease. Recently it was reported that the fruit inhibits passive cutaneous anaphylaxis and histamine release at mast cell. Type I immediate hypersensitivity of anaphylactic type is caused by released mediate chemical at mast cell. Histamine is also known as one of potent mediate chemical. Also release of mediate chemical is affected by specific stimulation of IgE combined with mast cell. Activation of mast cell is known to be stimulated by compound 48/80 and inhibited by increase of cAMP. In this experiment, the effect of water extract of Poncirus trifoliata Raf fruit (PT) on a histamine release, cAMP concentration and IgE production was measured. Compound 48/80 was administrated to the mouse peritoneal cell which was pretreated with PT. PT dosedependently inhibited histamine release at peritoneal mast cell and the serum level of histamine induced by compound 48/80. PT also instantly increased cAMP level of peritoneal mast cell right after it was added and the level gradually decreased. Production of IgE induced by antigens at mouse peritoneal cell was inhibited by PT. The IgE synthesis is induced by IL-4 and it is known that lipopolysaccharide(LPS) plus IL-4 cause an increase in IgE secretion by murine B cells. The effects of PT inhibited the production of IgE activated by LPS plus IL-4 at human U266B1 cells. These results indicate that PT has antiallergic activity by Inhibition of IgE production from B cells and histamine release by increase of cAMP.