• 대한본초학회지
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• 제23권2호
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• pp.87-97
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• 2008

Objectives : The present study was carried out to investigate the effect of Herba Polygalae Japonica on the proliferation and activation of eosinophils which were prepared from lung cells of asthma-induced mice by ovalbumin (OVA) treatment. Methods : C57BL/6 mouse was exposed to OVA three times a week for 6 weeks. The mouse lung tissues were dissected out, chopped and dessiciated with collagenase (1 ${\mu}g$/ml). Eosinophils were activated by rIL-3/rmIL-5 co-treatments. The lung cells were treated with extract of Herba Polygalae Japonica (EPJ), incubated for 48 hr at $37^{\circ}C$, and analyzed by flow cytometer, ELISA, RT-PCR and immunocytochemistry stain. Results : A significant cytotoxicity by drug treatment was not observed. The cell number ratio of granulocyte, CD3e-/CCR3+, CD3e+CD69+, CD4+, CD23+/B220+ cells was increased in rmIL-5/rIL-3 treated control group compared to the normal group. Cells numbers in the experimental animal group treated with EPJ was all decreased. In ELISA analysis, IL-4, IL-5, IL-13 levels and histamine release level were increased in the control group compared to the normal animal group, then significantly decreased in the experimental group with 100 ${\mu}g$/ml of EPJ treatment. In RT-PCR analysis, mRNA expressions of IL-4, IL-5, IL-13, CCR3 and eotaxin were increased in the control group compared to the normal animal group, then decreased in the experimental group with 100 ${\mu}g$/ml of EPJ treatment. And eosinophil proliferation levels were 18847${\pm}$,1527 (cpm) in the control group, 4676${\pm}$972 (cpm) in the positive control group, and 7709 ${\pm}$ 549 (cpm), 16839 ${\pm}$ 1403 (cpm), 16385 ${\pm}$ 1723 (cpm) in the experimental group with 100 ${\mu}g$/ml, 10 ${\mu}g$/ml, 1 ${\mu}g$/ml of EPJ treatment. Conclusions : The present data suggested that Herba polygalae japonica may have an effects on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of EPJ.

• Mycobiology
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• 제40권4호
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• pp.236-243
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• 2012

Pleurotus nebrodensis is an edible and commercially available mushroom in Korea. This study was conducted in order to evaluate the anticancer and immunopotentiating activities of crude polysaccharides, extracted in methanol, neutral saline, and hot water (hereafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of P. nebrodensis. ${\beta}$-Glucan and protein contents in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. nebrodensis ranged from 23.79~36.63 g/100 g and 4.45~6.12 g/100 g, respectively. Crude polysaccharides were not cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at a range of $10{\sim}2,000{\mu}g/mL$. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 11.76~27.06% in mice previously inoculated with sarcoma 180. Treatment with Fr. NaCl resulted in an increase in the numbers of spleen cells by 1.49 fold at the concentration of $50{\mu}g/mL$, compared with control. Fr. HW improved the immuno-potentiating activity of B lymphocytes through an increase in alkaline phosphatase activity by 1.65 fold, compared with control at $200{\mu}g/mL$. Maximum production of nitric oxide ($14.3{\mu}M$) was recorded in the Fr. NaCl fraction at $200{\mu}g/mL$. Production of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and interleukin-6 (IL-6) was significantly higher, compared to control, and IL-6 production was highest, in contrast to TNF-${\alpha}$, IL-$1{\beta}$, and positive control, concanavalin at the tested concentration of the various fractions. Results of the current study suggest that polysaccharides extracted from P. nebrodensis have a strong anticancer effect and may be useful as an ingredient of biopharmaceutical products for treatment of cancer.

• 한방안이비인후피부과학회지
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• 제18권1호
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• pp.116-134
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• 2005

Although the parallel prescription of Sopoongsangagambang (SG) administration along with external treatment such as spraying or ointment application on the skin is clinically used for the treatment of atopic dermatitis (AD), molecular mechanism underlying its effectiveness is unknown. Thus in the present study, diverse immune responses in terms of chemical mediators related to AD were investigated using an atopic mouse model NC/Nga after SG administration and external treatment (ET), and major findings are summarized as follows. 1. The clinical severities in 16 and 20 week old NC/Nga mice with SG and ET treatment were decreased to 72.2% and 62.3% respectively compared to the control NC/Nga mice with no drug treatment. 2. IgE, IL-4, IL-5, IL-6, IL-13, IgM, IgG1 and IgG2a levels in the serum of SG and ET treated NC/Nga mouse group were significantly decreased compared to the untreated control mice. In contrast, $IFN-{\gamma}$ showed a significant increase in the experimental group compared to the untreated control group. 3. The spleen weight of SG and ET treated NC/Nga mice was significantly decreased compared to the untreated control group. 4. The B/T ratio in the lymph node of SG and ET treated NC/Nga mice was increased compared to the untreated control group. $CD4^+\;and\;CD8^+$ cell numbers in the lymph node of SG and ET treated NC/Nga mice were significantly increased compared to the untreated control group, but $CD69^+\;and\;CD11a^+$ cells were significantly decreased. 5. mRNA expression levels of IL-4, IL-5, and CCR3 in the skin tissues of SG and ET treated NC/Nga mice were significantly decreased, and expression levels of IL-6, IL-13, $CD69^+/CD3{\varepsilon}^+\;and\;CD19^+/CD44^+$ in the skin tissues of SG and ET treated NC/Mga mice were significantly decreased compared to the untreated control group. $IFN-{\gamma}$ mRNA expression levels were increased compared to the untreated control group. 6. Histological observation of the ear and neck skin tissues showed that the extents of inflammation and infiltrated immune cells in the epidermis and dermis of SG and ET treated NC/Nga mice were highly reduced compared to the untreated control group. 7. Lymphokine assay showed a significant decrease in IL-4 levels in SG and ET treated NC/Nga mice compared to the untreated control group, but the levels of $IFN-{\gamma}$ secretion were significantly increased drug treated NC/Nga mice.

• 한국축산식품학회지
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• 제31권5호
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• pp.701-709
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• 2011

This study was performed to establish an effective extraction method of pig placenta extract that could be used for a putative functional food supplement with immunomodulatory effects. In the present study, we used different temperatures (4, 37, 60, 80, and $100^{\circ}C$) and different solvents (chloroform, NaOH, and phosphate buffered saline [PBS]) to extract the pig placenta. Among the different placenta extracts yielded by the different extraction methods, placenta extract (PE) in PBS at $80^{\circ}C$ for 30 min (referred to as PE-PBS80) showed a significant increase of nitric oxide production of up to 22.97 ${\mu}M/10^5$ cells at a 1 mg/mL dose (p<0.05 ) in J774A.1 cells than other extracts and control tested. Using PE-PBS80, further animal challenges were performed to identify the immune-enhanced effects. As a result, orally administered PE-PBS80 showed a significant increase in blood T and B cell activities and immunoglobulin (IgG and IgM) production. IgG and IgM levels increased to 41.53 mg/mL at a 20 mg dose on day 7 and to 27.38 mg/mL at a 10 mg dose on day 14, respectively (p<0.05). Furthermore, PE-PBS80 was also able to significantly enhance the immune modulator cytokine levels (p<0.05) compared to the control and vehicle treatments. Among the evaluated cytokines, the tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) level increased to 28.89 pg/mL at extract doses of 20 and 50 mg, the interleukin-$1{\beta}$ (IL-$1{\beta}$) level increased to 21.52 pg/mL at extract doses of 10, 20, 50 and 75 mg and the interferon (IFN)-${\gamma}$ level increased to 18.24 pg/mL at extract doses of 10, 20, and 50 mg. Therefore, this study presents an effective method for extracting pig placenta extracts and also demonstrates that pig placenta extracts had significant immunomodulatory effects not only at the cellular level but also in a mouse model, suggesting that this material could be used as an excellent candidate functional food supplement.

• 대한미생물학회지
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• 제13권1호
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• pp.55-62
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• 1978

항암제(抗癌劑)로 잘 알려진 cyclophosphamide(CY)를 성숙(成熟)한 마우스에 체중(體重) kg당(當) 300mg을 복강내(腹腔內)에 투여(投與)하여 CY가 체중(體重), 비장중량(脾臟重量) 및 말초임파조직(末梢淋巴組織)에 미치는 영향(影響)을 실험(實驗)하였다. CY는 마우스의 체중(體重)을 약간(若干) 감소(減少)시켰으나 체중(體重)은 곧 회복(回復)되었다. CY가 비장중량(脾臟重量)에 미치는 영향(影響)은 현저(顯著)하였는데 초기(初期)엔 비장중량(脾臟重量)은 감소(減少)되었으나 곧 회복(回復)되고 그 후(後) 정상(正常)보다 더 증가(增加)되었는데 그 증가(增加)는 CY투여(投與) 20일(日)이 지나서야 정상(正常)으로 되돌아왔다. 비장(脾臟) 및 임파절(淋巴節)의 조직학적(組織學的) 형태(形態)에 미치는 CY의 영향(影響)은 다양(多樣)하였다. 즉(卽), 흉선의존세포(胸線依存細胞)가 위치(位置)하는 비장(脾臟)의 periarterial lymphatic sheath와 임파절(淋巴節)의 paracortex는 골수유래세포(骨髓由來細胞)가 위치(位置)하는 follicles보다 1일(日) 내지(乃至) 2일(日) 늦게 소실(消失)되었으며 비장(脾臟)의 비대(肥大)가 최고(最高)에 달(達죠)하였을때 비장(脾臟)이나 임파절(淋巴節)의 구조(構造)는 동일(同一)한 임파양세포(淋巴樣細胞)로 구성(構成)된 interstitial tissue로 대치(代置)되어 있었다. 비장(脾臟)의 중량(重量)이 정상화(正常化)되어감에 따라 비장(脾臟)이나 임파절(淋巴節)의 구조(構造)가 정상화(正常化) 되어갔다. 이와 같은 본(本) 실험(實驗)의 소견(所見)은 CY는 골수유래세포(骨髓由來細胞) 뿐만 아니라 흉선의존세포(胸線依存細胞)에도 영향(影響)을 미친다고 사료(思料)된다. 저자(著者)들은 전보(前報)에서 CY에 의(依)한 지연성과민반응(遲延性過敏反應)의 항진(亢進)은 suppressor T cell의 제거(除去)에 기인(基因)한 것이라고 시사(示唆)하였는데 본(本) 실험결과(實驗結果)는 이를 뒷받침해준다고 사료(思料)되었다.

• 대한본초학회지
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• 제23권1호
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• pp.75-83
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• 2008

Objectives : The present study was carried out to investigate the effect of Hyperici Japonici Herba on the proliferation and activation of eosinophils which were prepared from lung cells of asthma-induced mice by ovalbumin(OVA) treatment. Methods : C57BL/6 mouse was exposed to OVA three times a week for 6 weeks. The mouse lung tissues were dissected out, chopped and dessiciated with collagenase(1${\mu}g$/ml). Eosinophils were activated by rIL-3/rmIL-5 co-treatments. The lung cells were treated with extract of Hyperici Japonici Herba(EHH), incubated for 48 hr at $37^{\circ}C$, and analyzed by flow cytometer. ELBA, RT-PCR, immunocytochemistry stain. Results : The cell number ratio of granulocyte, $CD3e^-$/$CCR3^+$, $CD3e^+$/$CD69^+$, $CD4^+$, $CD23^+$/$B220^+$ cells was increased in rmIL-5/rIL-3 treated control group compared to the normal group. Cells numbers in the experimental animal group treated with EHH was all decreased. In ELISA analysis, IL-4, IL-5, IL-13 protein levels and histamine release level were greatly increased in the control group compared to the normal animal group, then significantly decreased in the experimental group with 100 ${\mu}g$/ml of EHH treatment. In RT-PCR analysis, the HT value of IL-4, IL-5, IL-13, CCR3, Eotaxin were increased in the control group compared to the normal animal group, then decreased in the experimental group with 100 ${\mu}g$/ml of EHH treatment. And eosinophil proliferation levels were 18847${\pm}$1527(cpm) in the control group, 4676${\pm}$972(cpm) in the positive control group, and 8675${\pm}$159(cpm), 11352${\pm}$1005(cpm), 14325${\pm}$677(cpm) in the experimental group with 100 ${\mu}g$/ml, 10 ${\mu}g$/ml, 1 ${\mu}g$/ml of EHH treatment. Conclusions : The present data suggested that Hyperici Japonici Herba may have an effects on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of EHH.

• Archives of Pharmacal Research
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• 제29권10호
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• pp.911-920
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• 2006

Phenolic antioxidant butylated hydroxyanisole (BHA) is a commonly used food preservative with broad biological activities, including protection against chemical-induced carcinogenesis, acute toxicity of chemicals, modulation of macromolecule synthesis and immune response, induction of phase II detoxifying enzymes, as well as its undesirable potential tumor-promoting activities. Understanding the molecular basis underlying these diverse biological actions of BHA is thus of great importance. Here we studied the pharmacokinetics, activation of signaling kinases and induction of phase II/III drug metabolizing enzymes/transporter gene expression by BHA in the mice. The peak plasma concentration of BHA achieved in our current study after oral administration of 200 mg/kg BHA was around $10\;{\mu}M$. This in vivo concentration might offer some insights for the many in vitro cell culture studies on signal transduction and induction of phase II genes using similar concentrations. The oral bioavailability (F) of BHA was about 43% in the mice. In the mouse liver, BHA induced the expression of phase II genes including NQO-1, HO-1, ${\gamma}-GCS$, GST-pi and UGT 1A6, as well as some of the phase III transporter genes, such as MRP1 and Slco1b2. In addition, BHA activated distinct mitogen-activated protein kinases (MAPKs), c-Jun N-terminal kinase (JNK), extracellular signal-regulated protein kinase (ERK), as well as p38, suggesting that the MAPK pathways may play an important role in early signaling events leading to the regulation of gene expression including phase II drug metabolizing and some phase III drug transporter genes. This is the first study to demonstrate the in vivo pharmacokinetics of BHA, the in vivo activation of MAPK signaling proteins, as well as the in vivo induction of Phase II/III drug metabolizing enzymes/transporters in the mouse livers.

• 한국동물학회지
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• 제39권4호
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• pp.378-386
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• 1996

우리는 두 가지의 새로운 luciferase reporter plasmid를 개발하였는데 그 하나는 이 plasmid에 promoter조각을 삽입한 다음에 그 promoter의 활성을 측정하는 용도로 사용 될수 있고, 다른 하나는 매우 낮은 basal promoter 활성을 갖고 있기 때문에 진핵생물의 전사 조절인자의 연구에 도움이 될 수 있다. 후자의 reporter plasmid에는 17염기쌍의 initator 와 Spl,GAL4그리고 일부 Drosophila homeodomain protein의 결합우뷔에 해당하는 cis elements등을 들어 있다.그리고 tranciption termination을 촉진할 수 있는 signal을 initiator앞서 삽입하여 이런 reporter plasmid에 존재할 수 있는 cryptic promoter에서 시작된 transcript가 luciferase reporter cDNA로 진행되는 것을 방지하는 개선된 reporter plasmid를 제조하여 promoter활성을 Drosophila Schneider line 2 cells을 이용한 transient transfection assay방법으로 측정하였다. 여기에 사용한 termination 촉진 signal은 SV40 polyadenylation signal의 3연속 조각(AAA)과 tenscription termination signal이 포함된 것으로 믿어지는 mouse c-mos 유전자의 일부조각 (UMS)이다. 기대한으로 이 두가지의 signal을 삽입하였을 때 basal promoter활성이 최대한으로 감소하였으며 이 두 가지 signal을 삽입된 reporter plasmid를 사용하여 promoter의 활성을 보다 sensitive하게 측정하였다. 이 reporter plasmid는 Droiophlla melanogaiter뿐만 아니라 포유동물을 포함한 고등생물의 전사 조절인자 연구의 한 도구로 사용될 수 있을 것이다.

• 대한예방한의학회지
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• 제6권2호
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• pp.156-167
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• 2002

This experimental study was carried out to evaluate the effects of Kimchi intake of Cordyceps sinensis extract (CDSE) supplementation on cytokine-induction and immune response in mice. To study in experiments using male Balb/c mice fed Kimchi and Kimchi of CDSE supplementation (addition of 2% of total Kimchi weight) containing fed experimental diet during 2 weeks. Experimental mice were fed control diet or diet containing freeze-dried Kimchi at the level of 5%(w/w) or 5% freeze-dried Kimchi with 2% CDSE supplementation. The main ingredient of Kimchi was Korean cabbage and fermentation was carried out at $4^{\circ}C$ for three weeks. Freeze-dried 2% CDSE supplementation was added to Kimchi at the beginning of fennentation. In order to investigate the effect of Kimchi intake of CDSE supplementation (5%Kimchi-2%CDSE), the following was performed; body weight, food intake, hematological parameter, serum level of mouse interleukin-4 (mIL-4) and mouse $interferon-{\gamma}$ $(mIFN-{\gamma})$, and, the percentage of CD3+/CD4+, CD3+/CD8+, B220+ in splenic cells. The results of final body weight, and food diet intake of two Kimchi groups were lower than those of the control group (not supplemented experimental diet). The hematology change obtained from the level of WBC (white blood cell) and platelet were not affected by feeding different dietary regiments, but the level of RBC (red blood cells) HB (hemoglobin), and spleen weight of two Kimchi groups were increased significantly than those of the control group. The serum level of IL-4 and $IFN-{\gamma}$ of two Kimchi groups were increased significantly than those of the control group, also enhanced the percentages of the CD3+/CD4+ and CD3+/CD8+ by 5% freeze-dried Kimchi, and 5%Kimchi-2%CDSE group were 43.9 and 60.1%, and 96.0 and 174% than those of the control group, respectively. From these results, it can be concluded that Kimchi itself has an immuno-stimulatory effect and Kimchi contaning 2% CDSE supplementation has the more pronounced effect in vivo system.

• Laboraroty Animal Research
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• 제34권4호
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• pp.166-175
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• 2018

Recombination activating gene-2 (RAG-2) plays a crucial role in the development of lymphocytes by mediating recombination of T cell receptors and immunoglobulins, and loss of RAG-2 causes severe combined immunodeficiency (SCID) in humans. Rag-2 knockout mice created using homologous recombination in ES cells have served as a valuable immunodeficient platform, but concerns have persisted on the specificity of Rag-2-related phenotypes in these animals due to the limitations associated with the genome engineering method used. To precisely investigate the function of Rag-2, we recently established a new Rag-2 knockout FVB mouse line ($Rag-2^{-/-}$) manifesting lymphopenia by employing a CRISPR/Cas9 system at Center for Mouse Models of Human Disease. In this study, we further characterized their phenotypes focusing on histopathological analysis of lymphoid organs. $Rag-2^{-/-}$ mice showed no abnormality in development compared to their WT littermates for 26 weeks. At necropsy, gross examination revealed significantly smaller spleens and thymuses in $Rag-2^{-/-}$ mice, while histopathological investigation revealed hypoplastic white pulps with intact red pulps in the spleen, severe atrophy of the thymic cortex and disappearance of follicles in lymph nodes. However, no perceivable change was observed in the bone marrow. Moreover, our analyses showed a specific reduction of lymphocytes with a complete loss of mature T cells and B cells in the lymphoid organs, while natural killer cells and splenic megakaryocytes were increased in $Rag-2^{-/-}$ mice. These findings indicate that our $Rag-2^{-/-}$ mice show systemic lymphopenia with the relevant histopathological changes in the lymphoid organs, suggesting them as an improved Rag-2-related immunodeficient model.