• 대한본초학회지
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• 제31권4호
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• pp.79-85
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• 2016

Objectives : The aim of this study is to investigate anti-atopic dermatitis effect using ato-tang.Methods : Ato-tang was external treatment to NC/Nga mice for 4 weeks, where atopic dermatitis was induced by DNCB at 1% and 0.4% for 3 weeks. Atopic dermatitis index score was measured using eye observation and picture evaluation. The histopathological change of dorsal skin was observed by hematoxylin and eosin (H&E) staining. Cytokines including IL-4, IL-5 and IL-13 were measured by Luminex or reverse transcriptase polymerase chain reaction (RT-PCR) analysis and immunoglobulin E (IgE) was measured by ELISA reader.Results : The dorsal skin of Ato-tang group showed decrease in erythema, pruritus, dry skin, edema, excoriation, erosion and lichenification level through naked eye observations. Immunoglobulin cell infiltration and the thickness of epidermis were significantly decreased in the dorsal skin compared to control. Production of Th2 cytokines (IL-4, IL-5, IL-13) and IgE level in serum were all significantly decreased, in comparison with control. In addition, mRNA expression level of Th2 cytokines (IL-4, IL-5, IL-13) in spleen was decreased, in comparison with control.Conclusion : The results indicated that external treatment of ato was improved skin barrier function in the symptoms of atopic dermatitis disease. Also, atopic dermatitis factors where cytokine as well as immunoglobulin E in serum and mRNA expression were decreased, respectively, in comparison with control. Therefore, we suggest that ato could be effectively used as a external therapeutic drug based on atopic dermatitis factors.

• 동의생리병리학회지
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• 제19권4호
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• pp.922-927
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• 2005

In the present study, baicalin, baicalein, and wogonin, a major flavone isolated from Scutellaria Radix were examined for their effects on PMA-induced Interlukin-6 (IL-6), $interferon-\gamma(IFN-\gamma)$, tumor necrosis factor $(TNF)-\alpha$, IL-4, IL-10, and IL-13 productions in the PMA-stimulated $CD4^+$ Jurkat T cells. These three compounds inhibited PMA-induced Th1 cytokine $(IL-6,\;IFN-\gamma,\;TNF-\alpha)$ and Th2 cytokine (IL-4 and IL-13) productions in a concentration-dependent manner. But wogonin, but not baicalin baicalein, increased PMA-induced IL-10 production. These results suggest that baicalin, baicalein, and wogonin, a major flavone modulate Th1 and Th2 cytokine productions in $CD4^+$ Jurkat T cells and these properties may contribute to the anti-atopic dermatitis activity of Scutellaria Radix.

• KSBB Journal
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• 제24권4호
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• pp.383-392
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• 2009

아토피피부염 (AD)은 천식, 음식 알레르기, 비염 같은 전신아토피질환을 동반하는 만성재발성 피부염증질환이다. 아토피피부염과 관련된 IL-17의 임상적 역할은 다양한 조건에서 보고되고 있으며, 또한 건선 피부 상태에 깊숙이 관여하고 있다. IL-17은 각질세포 (keratinocytes)에서 과잉으로 생산되며, 아토피피부염의 말초임파구에서도 다량 생성됨을 세포내염색을 통하여 확인된 바 있다. 본 연구에서는 창이자 추출물 (XS-E와 XS-FL)이 NC/Nga 생쥐의 $CD4^+$ T 세포에서 유도된 Th17 세포의 분화억제 및 IL-17의 생산량 감소 효과에 대한 실험을 하였다. 그 결과 XS-E와 XS-FL을 처리한 섬유아세포에서 세포독성은 나타나지 않았고, 4일간 XS-E와 XS-FL에 동시배양 한 $CD4^+$ T 세포의 IL-17 생산량을 FACS로 분석한 결과 $100\;{\mu}g$/mL XS-E 처리군의 IL-17 생산량은 32.3%로 대조군에 비하여 2배 이상의 감소를 나타내었으며, $20\;{\mu}g$/mL XS-30% AFL (acetone XS-FL) 처리군의 Th17 세포는 19.6%으로 대조군에 비하여 3.5배 억제되었다. 또한 real-time PCR을 이용하여 IL-17A와 IL-22 mRNA의 유전자 발현량을 비교 분석한 결과, IL-17A와 IL-22 mRNA의 유전자발현의 RQ값은 XS-E와 XS-30% AFL를 처리한 실험군이 대조군에 비하여 유의성 있는 감소를 나타내었다(p < 0.01, p < 0.001). ELISA로 측정한 IL-17A 생산량은 XS-E와 XS-30% AFL를 처리한 실험군이 대조군에 비하여 현저히 감소 (p < 0.05, p < 0.001)하였다. Th17 세포의 증식을 알아보기 위하여, rIL-6와 TGF-$\beta$로 분화시킨 Th17 세포를 CFSE로 표지한 후 rIL-23 처리를 하여 4일간 배양하여 증식을 유도시켰다. 대조군의 Th17 세포 분열은4일 동안 4번에 걸쳐 비슷한 세포수의 증식이 일어나는 것을 CFSE를 통하여 확인하였고, XS-30% AFL 처리군은 CFSE의 형광 분포가 점점 감소하여 Th17 세포의 증식이 억제됨을 알 수 있었다.

• BMB Reports
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• 제50권12호
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• pp.640-646
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• 2017

Regulatory B cells, also well-known as IL-10-producing B cells, play a role in the suppression of inflammatory responses. However, the epigenetic modulation of regulatory B cells is largely unknown. Recent studies showed that the bromodomain and extra-terminal domain (BET) protein inhibitor JQ1 controls the expression of various genes involving cell proliferation and cell cycle. However, the role of BET proteins on development of regulatory B cells is not reported. In this study, JQ1 potently suppressed IL-10 expression and secretion in murine splenic and peritoneal B cells. While bromodomain-containing protein 4 (BRD4) was associated with $NF-{\kappa}B$ on IL-10 promoter region by LPS stimulation, JQ1 interfered the interaction of BRD4 with $NF-{\kappa}B$ on IL-10 promoter. In summary, BRD4 is essential for toll like receptor 4 (TLR4)-mediated IL-10 expression, suggesting JQ1 could be a potential candidate in regulating IL-10-producing regulatory B cells in cancer.

• IMMUNE NETWORK
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• 제5권1호
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• pp.16-22
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• 2005

Background: IL-18 was originally cloned as a IFN-${\gamma}$ inducing factor in primed T cells. In synergy with IL-12, IL-18 has been shown to induce strikingly high levels of IFN-${\gamma}$ production by T cells and to enhance Th1 development. Also this cytokine exerts induction of Th2 development through IL-4 induction. Methods: Resting $CD4^+$ T cells were sorted by negative selection and activated by anti-CD3 plus anti-CD28 Ab. Expression of IL-12 binding sites, IL-18 binding sites, IL-18R ${\alpha}$, and GATA-3 mRNA were analysed by FACS and RT-PCR, respectively. Results: Resting $CD4^+$ T cells expressed IL-18R ${\alpha}$ chain but not IL-18 binding sites, suggesting a lack of IL-18R ${\beta}$ expression. IL-18R ${\alpha}$ was maintained on the Th1 and Th2 committed cells. IL-18 binding sites were induced on the Th1 but not Th2 cells. Exposure of these cells to IL-18 led to up-regulation of GATA-3 mRNA expression only in Th2 committed cells. To elucidate the relationship between IL-18R ${\alpha}$ expression and GATA-3 induction by IL-18, Th1 and Th2 committed cells were further cultured in medium with or without IL-12 for 2 days. IL-12 binding sites were maintained on the Th1 and Th2 cells regardless of IL-12 treatment, but IL-18R a expression was rapidly down-regulated on the IL12-untreated Th2 cells which did not induce GATA-3 mRNA expression followed by IL-18 stimulation. Conclusion: IL-12 supports expression of IL-18R ${\alpha}$ and GATA-3 mRNA expression was induced by IL-18 through IL-18R ${\alpha}$ without expression of IL-18 binding site in Th2 cells.

• 대한한방내과학회지
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• 제25권3호
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• pp.408-417
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• 2004

Objective : The aim is to identify the effects of Glycyrrhiza uralensis Fisch on immunocyte and cytokine production in asthmatic laboratory mice. This experiment was designed to investigate the antiallergic and antiinflamatary the effects of Glycyrrhiza uralensis Fisch on asthma. Materials and Methods : We measured the eosinophil, IL-4, IL-5, IL-13, $IFN-{\gamma}$, CD4, CDS, CD69, CCR3, CD11b, Gr-1 in bronchoalveolar lavage fluid of ovalbumin induced asthmatic mice. Results : Glycyrrhiza uralensis Fisch increased the proliferation of eosinophils, IL-4, IL-5, IL-13, IgE, granulocyte, CCR3, CD4, IgE, CD69. Glycyrrhiza uralensis Fisch increased the proliferation of $IFN-{\gamma}$. Conclusion : Results suggest that Glycyrrhiza uralensis Fisch extract is useful in treatment and prevention allergic asthma.

• 대한한방소아과학회지
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• 제32권3호
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• pp.1-15
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• 2018

Objectives Alismatis Rhizoma has been known to suppress inflammation and allergic reaction. However, the cellular target of Alismatis Rhizoma and its mechanism of action remain unclear. This study was designed to examine the effect of Alismatis Rhizoma extract (ALC) on the RBL-2H3 mast cells in vitro and on the OVA/alum sensitized mice ex vivo. Methods In the study, RBL-2H3 mast cells were cultured in minimal essential medium (MEM) for 24 hours, and treated separately with cyclosporin A and varying doses of ALC, and then stimulated with Phorbol 12-myristate 13-acetate (PMA) (50 ng/ml) and Ionomycin ($0.5{\mu}M$). The levels of IL-13, IL-4 were measured by ELISA analysis. The mRNA levels of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$ were analyzed with Real-time PCR. Also, manifestations of MAPKs transcription factors and $NF-{\kappa}B$ p65 translocation were analyzed by western blotting in vitro. Subsequently, for ex vivo experiment, we induced allergic inflammation on Balb/c mice by OVA/alum and administered ALC orally. And we measured serum OVA-specific IgE level and IL-4, IL-13 in the splenocyte culture supernatant by ELISA analysis. Results ALC was shown to suppress mRNA expression of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$, and to inhibit the IL-13, IL-4 production. Also ALC reduced an activation of mast cells specific signal MAPKs transcription factors and $NF-{\kappa}B$ p65 from the western blot analysis in in vitro experiment. In ex vivo, ALC oral adminstration decreased the level of OVA-specific IgE in serum, and IL-4, IL-13 in the splenocyte culture supernatant. Conclusions ALC is shown to reduce inflammation and allergic response by suppressing Th2 cytokines through the regulation of transcription factors MAPKs and $NF-{\kappa}B$ p65 in mast cells. Administration of ALC suppressed OVA-specific IgE in ovalbumin allergy model through the inhibition of Th2 cytokine. In conclusion, ALC can be considered as an effective treatment for allergic diseases such as atopic dermatitis.

• 사상체질의학회지
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• 제12권1호
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• pp.201-215
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• 2000

1. 연구배경 및 목적 사상의학(四象醫學)에서 열다한소탕(熱多寒少湯)은 태음인(太陰人)의 뇌경색증 급성기에 매우 빈번하게 사용되는 처방 중 하나이다. 그래서 저자는 열다한소탕(熱多寒少湯) 투여에 따른 뇌경색증 환자의 면역계 특히 세포활성문질만계의 변화에 대해 연구하기로 하였다. 2. 방법 원광대학교 광주한방병원 사상의학과에 입원한 뇌경색증 한자 8인(남자 3명, 여자 5명)을 대상으로 하여, 내원 당시와 2-4주간의 열다한소탕 투여 후의 세포활설물질망계의 변화를 추적하였다. 또한 정상태음인의 말초혈액단핵구를 채취하여 리포다당체 처리시의 변화를 열다한소탕(熱多寒少湯) 전처리 유무에 따라 비교 분석하였다. 3. 결과 IL-2 혈장 농도는 정상 대조군보다 환자군에서 더 낮았다. $IFN-{\gamma}$의 경우 역시 정상 대조군보다 약간 낮았으나 통계학적 유의성은 없었다. IL-4, IL-6, IgE의 혈장중 수준은 정상 대조군에 비해 환자군에서 현저히 상승되어 있었다. 그러나, 2-4주간의 열다한소탕(熱多寒少湯) 투여후 $IFN-{\gamma}$, IL-2의 혈장 중 수준은 현저히 상승되었고, IL-4, IL-6, Ig E 수준은 현저히 억제되었다. 한편 정상 태음인의 말초혈액단핵구에 리포다당체 처리를 한 후 $IFN-{\gamma}$, IL-2, IL-6가 상승되었으나, 열다한소탕(熱多寒少湯) 전처리를 한 경우에서는 이들 물질의 생성이 유의하게 억제되었다.

• Parasites, Hosts and Diseases
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• 제34권3호
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• pp.185-190
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• 1996

폐흡층(Paragonimus westermani) 감염시 일어나는 TH cytokin 반응을 알아보고자 마우스에 폐흡충 피낭유충을 감염시킨 후 비장세포를 Con A로 자극하여 TH1-specific cytokine인 $IFN-{\gamma}$와 TH2-specific cytokin인 IL-4의 생산량을 감염시기별로 효소표식 면역검사법으로 측정하였다. 폐흡충 감염 마우스의 비장세포에서 생산되는 IL-4는 감염 후 3일($410{\;}{\pm}{\;}60.9{\;}pg/ml$)에 최고치에 도달한 후 2주($343{\;}{\pm}{\;}59.0{\;}pg/ml$)까지 대조군에 비해 높게 유지되었으나 감염 후 4주에는 감소되기 시작하여 6주에는 대조군의 생산량과 비슷하였다. 한편 폐흡충 감염 마우스의 비장세포에서 생산되는 $IFN-{\gamma}$는 감염 후 1주($151{\;}{\pm}{\;}32.2{\;}pg/ml$)에만 대조군에 비해 높게 증가되었을 뿐 2주부터 감소되기 시작하여 감염 후 6주에는 전혀 측정되지 않아 오히려 대조군의 생산량보다도 적었다. 또한 폐흡충 감염 마우스의 혈청 내 IL-4의 양은 감염 후 4주부터 6주에 대조군에 비해 높게 증가되었으며 혈청내 $IFN-{\gamma}$의 양은 전 실험기간을 통해 측정되지 않았다. 이상의 결과를 종합할때 폐흡층 감염마우스에서는 $IFN-{\gamma}$보다는 IL-4가 증가되는 TH2 cytokine 반응이 주로 일어남을 알 수 있었다.

• 대한바이러스학회지
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• 제28권1호
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• pp.71-78
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• 1998

Vaccinia virus is the prototype orthopoxvirus that has been used as a vaccine strain for small pox. This virus has been used to express a variety of cellular and viral genes in mammalian cells at high levels. Interleukin-4 (IL-4) has been found to stimulate the proliferation of T cells and enhance the cytolytic activity of cytotoxic T lymphocytes. To test the immunotherapeutic potential of IL-4 delivered in vivo by poxvirus, a recombinant vaccinia virus expressing the murine IL-4 gene (RVVmIL-4) was constructed. A high level of IL-4 production was confirmed by infecting HeLa cells and measuring IL-4 in cell culture supernatant by ELISA. As a tumor model, two cell lines were used; the murine T leukemic line P388 and the murine breast cancer line TS/A. CDF1 mice were intraperitoneally inoculated with $1\;{\times}\;10^5$ cells of P388. Mice were injected at the same site with $5\;{\times}\;10^5\;PFU$ of recombinant vaccinia virus; first, 3 days after the injection of tumor cells and thereafter once every week for 3 weeks. Intraperitoneal injections of RVVmIL-4 significantly prolonged the survival time of mice inoculated with tumor cells. All mice injected with RVVmIL-4 remained alive for 30 days after the postinoculation of tumor cells, while 100% and 70% of the animals injected with saline or wild type vaccinia virus died, respectively. In another tumor model using TS/A, tumor was established by subcutaneously inoculating $2{\times}10^5$ tumor cells to BALB/c mice. After tumor formation was confirmed on day 4 in all mice, $5\;{\times}\;10^6\;PFU$ of RVVmIL-4 was inoculated subcutaneously three times, once every week for 3 weeks. The TS/A tumor was eradicated in two of the nine mice. Seven of the nine mice treated with RVVmIL-4 developed a tumor, but tumor growth was significantly delayed compared to those treated with saline or wild type vaccinia virus. These results indicate that recombinant vaccinia viruses may be used as a convenient tool for delivering immunomodulator genes to a variety of tumors.