• 한국수의병리학회지
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• 제3권1호
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• pp.15-25
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• 1999

To elucidate the mechanism of age-related development in FGS/NgaKIST mice with spontaneous glomerulosclerotic lesion, we examined expression and localization of various cytokine mRNA in the kidney in the progression of diseases. This mouse model is the first to develop spontanously occuring glomerosclerotic lesion in the kidney. In this study, we detected the up-regulation of local cytokine genes such as IL-1$\beta$, IL-2, IL-6, IL-10, TNF-$\alpha$, TGF-$\beta$, and IFN- $\gamma$ in the kidneys. In RT-PCR and Southern blot analysis, we detected gradual expressions of cytokine mRNA of IL-1$\beta$, IL-2, IL-6, IFN- $\gamma$, and TNF $\alpha$ mRNA during the course of disease. Other cytokines including IL -10 and TGF -$\beta$ were found to be appeared the slightly expressed level at 3 to 12 weeks before onset of inflammatory lesion but they are highly expressed at the end-stage of the disease accompaning high proteinurea and wasting. In situ RT-PCR, each cytokine mRNA were specifically localized in a variety of cells including mesangial, endothelial, parietal epithelial, tubular epithelial, arterial muscle cell, and infiltrated inflammatory cells. In addition, TNF - $\alpha$was detected moderately in the visceral and parietal epithelial cell, but weakly in endothelial and mesangial cells, whereas IL-1 $\beta$ and IL -6 were strong in mesangial regions. IL-6 and TNF- $\alpha$ was highly localized in the damaged proximal and collecting tubules. Especially, TGF -$\beta$ mRNA was highly found in mesangial cells within glomerulus and interstitium during the end-stage of this disease.. These results indicate that pro inflammatory cytokines such as IL-1 $\beta$, IL-2, IL-6, and TNF- $\alpha$ were gradually expressed from the early stage of this disease to the end-stage, and that IL-10 and TGF-$\beta$ may be important in the accumulation of extracellular matrix(ECM) within glomerulus and periglomerular fibrosis in the progression of this disease as well as tissue destruction in end-stage of this disease.

• 대한한의학회지
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• 제24권2호
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• pp.166-178
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• 2003

Objectives : Nitric oxide (NO) plays an important role in normal and pathophysiological cells as a messenger molecule, neurotransmitter, microbiological agent, or dilator of blood vessels and arteriosclerosis, respectively. This study was undertaken to understand the mechanism of NO production and effect of Hyeolbuchukeo-tang (Xiefuzhuyu-tang) on NO production in cultured vascular smooth muscle cell (VSMC). Methods and Results : VSMC was isolated from aorta and cultured. Cultured primary cells were identified as VSMC with anti--smooth muscle actin antibody. A large amount of NO was produced in cultured VSMC treated with $IFN-{\gamma}$ plus TNF in a time- and dose-dependent manner. $TNF-{\alpha}$ was a more efficient stimulator than $IFN-{\gamma}$ in NO production of cultured VSMC. iNOS protein wasdetected within 3 hrs and it increased up to 12 hrs in a time-dependent manner. However, accumulated NO in cytokine-treated VSMC was not detected within 3 hrs. NO production in cytokine-treated VSMC showed the dose- and time-dependent manner, and increased up to 48 hrs. The activated VSMC produced a large amount of NO (about 60 uM). Hyeolbuchukeo-tang (Xiefuzhuyu-tang) alone did not induceNO production, but it potentiated the effect of $TNF-{\alpha}$ on NO production and increased NO production by about 20%. Hyeolbuchukeo-tang (Xiefuzhuyu-tang) did not affect the transcriptional activity of iNOS gene, but increased the accumulation of iNOS. These results indicate that Hyeolbuchukeo-tang (Xiefuzhuyu-tang) could modulate the translational level of iNOS. PKC did not modulate NO production, but calcium ionophore A23187 decreased NO production. However, Hyeolbuchukeo-tang (Xiefuzhuyu-tang) elevated the decreased NO production in A23187-treated VSMC by modulating the stability of iNOS transcripts. Half-life of the synthesized transcripts appeared to have about 6 hrs. PDTC, an $NF-{\kappa}B$ inhibitor, blocked the accumulation of iNOS mRNA, indicating that $NF-{\kappa}B$ served as an important modulator in the transcriptional regulation of iNOS. As Hyeolbuchukeo-tang (Xiefuzhuyu-tang) potentiated the effect of the $TNF-{\alpha}$ on NO production but had no additional effect on PDTC-modulated NO production, it is suggested that Hyeolbuchukeo-tang (Xiefuzhuyu-tang) enhances the $TNF-{\alpha}-mediated$ NO production of VSMC by modulating the iNOS activity and the stability of iNOS transcripts in activated VSMC having the elevated intracellular calcium ion. Conclusions : This study suggests that Hyeolbuchukeo-tang (Xiefuzhuyu-tang) has a potential capacity for preventing and treating diseases of the circulation system, including arteriosclerosis.

• 대한한의학회지
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• 제30권5호
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• pp.61-76
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• 2009

Background: Membranous nephropathy (MN) is the most common cause of adult nephrotic syndrome worldwide and has been defined as granular subepithelial deposition of immune complexes along the glomerular basement membrane (GBM). MN has few known treatments and gives rise to side effects under treatment with steroids and immunosuppressives. Objective: The purpose of this experimental study was to demonstrate the effects of Scutellariae Radix extract (SRE) treatment on MN mouse model induced by cBSA. Methods: We divided mice into 4 groups. The Normal group had no treatment. We induced MN mouse model to the other 3 groups by injecting cBSA into the abdominal cavity. The control group was treated with cBSA (10 mg/kg, i.p.) only. The second group, 'SRE-250', was treated with cBSA (10 mg/kg, i.p.) and SRE (250 mg/kg, p.o.). The third group, 'SRE-500', was treated with cBSA (10 mg/kg, i.p.) and SRE (500 mg/kg, p.o.). After cBSA and SRE treatment for 4 weeks, gain in body weight, 24hrs proteinuria, serum albumin, total cholesterol, triglyceride, BUN and creatinine of all groups were measured. TNF-$\alpha$, IL-6, IL-1$\beta$, IL-10, IFN-$\gamma$, IgA, IgM and IgG levels of all groups were gauged. H&E staining and electron microscopy of the kidney were observed. Results: SRE showed significant decrease in the 24hrs proteinuria, serum triglyceride, BUN, TNF-$\alpha$, IL-6, serum IgA, IgM and IgG levels compared with the control group. SRE showed increase in the serum IL-10 and IFN-$\gamma$ levels compared with control on RT-PCR. SRE considerably decreased in the thickening of the GBM on H&E staining and deposition of electron-density on electron microscopy of the kidney compared with the control. Conclusions: According to the above results, it is suggested that SRE decreases the symptoms of MN induced by cBSA in mouse model. Therefore, SRE seems to be applicable to MN in clinical practice.

• 식품산업과 영양
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• 제9권1호
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• pp.36-45
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• 2004

Bamboo salt has been used for the purpose of prevention and treatment of various diseases in Korea. Present study was carried out to ascertain the effects of purple bamboo salt upon anti-allergic effect, anti-inflammatory activity and immune-enhance effect as well. Purple bamboo salt significantly inhibited the ear swelling response and histamine release induced by compound 48/80 in mice and rat peritoneal mast cells. Purple bamboo salt (0.01 ∼ lg/kg) also dose-dependently inhibited the passive cutaneous anaphylaxis by oral administration. Purple bamboo salt (1 mg/mL) in hibited phorbol 12-myristate 13-acetate plus calcium ionophore A23187-stimulated tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-1${\beta}$ and IL-6 secretion, by 67.04${\pm}$0.08%, 68.01${\pm}$1.85%, 69.48${\pm}$0.54%, respectively. In addition, purple bamboo salt inhibited the expression of TNF-${\alpha}$ mRNA in HMC-1 cells. Finally, we investigated the effect of purple bamboo salt in the forced swimming test (FST) and the change of purple bamboo salt-mediated cytokine production from MOLT-4 cells. At the 7th, immobility time was significantly decreased in the purple bamboo salt-administration group (35.4 ${\pm}$5.9 s for 1 g/kg) in comparison with the control group (93.2 ${\pm}$ 15.45). After FST, the content of glucose in the blood serum was increased and the levels of blood urea nitrogen, lactic dehydrogenase was decreased in purple bamboo salt-administration group. However, it had no effect on the elevation of CK and TP level. Purple bamboo salt (1 mg/mL) significantly increased the interferon (IFN)-${\gamma}$ and IL-2 level compared with media control (about 3.7-fold for IFN-${\gamma}$, about 3.5-fold for IL-2, p〈0.05) but did not affect the IL-4.

• Asian-Australasian Journal of Animal Sciences
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• 제21권11호
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• pp.1651-1658
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• 2008

This study was conducted to determine the effects of dietary supplementation of either 100 mg/kg chito-oligosaccharide (COS) or chlortetracycline (CTC) with corn-soybean-fish meal on immunity in broiler chickens. A total of 147 one-day old male broiler chicks were randomly allocated to 3 treatments with 7 replicate pens per treatment and 7 birds per pen. The experimental diets consisted of a control diet based on corn, soybean and fish meal without COS and any antibiotic supplement and similar diets supplemented with either CTC (80 mg/kg from d 1 to 21 and 50 mg/kg from d 22 to 42) or COS (100 mg/kg from d 1 to 42). During the entire experimental period, all birds had ad libitum access to diets and water. The main immune organ indices, T-lymphocyte proliferation, serum cytokine concentrations, serum NO level and serum iNOS activity were measured on d 21 and d 42. On d 21, broilers fed 100 mg/kg COS had improved (p<0.01) indices of spleen, thymus, and bursa of Fabricius compared with the control and CTC birds. Birds receiving 100 mg/kg COS had higher (p<0.05) serum concentrations of $IL-1{\beta}$, IL-6, IgM, NO and iNOS than birds on the control treatment. Serum $Ca^{2+}$ level of birds fed 100 mg/kg COS tended to be higher (p = 0.049) than in birds fed CTC. On d 42, the birds fed 100 mg/kg COS had higher (p<0.05) concentrations of TNF-${\alpha}$ and IgM in serum than birds in both the CTC and control treatments. Birds fed 100 mg/kg COS had a higher concentration of IFN-$\gamma$ than the control group. In conclusion, dietary supplementation of COS appeared to improve the immunity of broilers by promoting the weight of the main immune organs, increasing IgM secretion, stimulating microphages to release $TNF-{\alpha}$, $IL-1{\beta}$, IL-6 and IFN-$\gamma$, and activating iNOS to induce NO.

• Journal of Periodontal and Implant Science
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• 제29권4호
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• pp.803-823
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• 1999

In order to reveal immunopathogenesis of periodontal tissue destruction, it is important to clarify the molecular mechanism of trafficking and retention of activated leukocytes, including monocytes/macrophages. Gingival fibroblasts may be involved in the regulation of inflammatory cell accumulation in the extravascular periodontal connective tissues via cytokine production and surface expression of adhesion molecules. In this study, it was investigated the molecular basis for the adhesive interactions between monocytes and fibroblasts such as peri-odontal ligament fibroblast(PDLF), human gingival fibroblast(HGF), and human dermal fibroblast(HDF). First, it was examined the evidence whether monocyte-fibroblast cell contact may cause signal transduction in fibroblasts. Being directly in contact with fixed human monocyte cell line THP-1, or U937, upregulation of IL-6 production, $TNF-{\alpha}$ mRNA expression and increased cell proliferation could be seen for fibroblasts. IL-6 production induced by monocyte- fibroblast coculture were further increased when fibroblasts had been pretreated with $IFN-{\gamma}$ or $IL-1{\beta}$ , and monocytes with LPS. Next, it was examined the expression of ICAM-1 which has been known to be involved in accumulation and activation of leukocytes in inflammatory diseases such as periodontitis. ICAM-1 was upregulated up to 10-fold on PDLF, HGF, and HDF by exposure to $IFN-{\gamma}$ or $IL-1{\beta}$. Furthermore, anti-ICAM-1 monoclonal antibody clearly blocked cocultureinduced IL-6 production by fibroblasts, suggesting that $ICAM-1/{\beta}_2$integrin pathway is involved in periodontal fibroblastmonocyte interaction. Overall, these findings provide evidence that periodontal fibroblasts could be involved in the accumulation and retention of monocytes/macrophages in periodontal inflammatory lesion at least in part by ICAM-1 expression. In addition, periodontal fibroblast-monocyte interaction could cause activation signals in fibroblasts intracellularly which result in cytokine production and cell proliferation. Thus, periodontal fibroblasts are speculated to play an important role in immunoregulation and tissue destruction in chronic periodontal diseases by interaction with monocytes/macrophages.

• 한국가금학회지
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• 제43권3호
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• pp.135-142
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• 2016

본 연구는 4종의 재래닭인 연산오계, 현인흑계, 횡성약닭, 황봉의 일반성분, 카르노신, 안세린 함량 및 면역활성을 비교하기 위하여 실시하였고, 대조구로 백색 레그혼을 사용하였다. 시험용 닭을 모두 동일한 조건에서 13주간 사육하여 도계한 후 화학적 성분조사를 실시하였다. 일반성분 분석에서는 4종의 재래닭이 백색레그혼에 비하여 조지방 함량이 높았으며, 콜레스테롤 함량은 현인흑계가 다른 품종에 비하여 높았고 횡성약닭이 유의적으로 낮았다(p<0.05). 면역증진 효능을 측정하기 위하여 대식세포를 이용하여 실험한 결과는 재래닭 4종의 가슴육에서만 대식세포 증식 및 NO 억제 효능이 있었다. 대식세포 탐식작용은 현인흑계와 횡성약닭가슴육에서 유의적으로 증가되었고, 염증 관련 사이토카인(IL-4, IL-10, $IFN-{\gamma}$) 유전자 발현은 백색레그혼에 비하여 4종의 재래닭에서 억제효과가 컸다. 결론적으로 4종류의 재래닭이 백색레그혼에 비하여 면역증진 효능이 높았으며, 특히 가슴육에서 효능이 있었다. 재래닭 품종 간에도 일반성분 및 면역조절능에 차이가 있는 것으로 나타났다.

• 한국임상수의학회지
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• 제26권5호
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• pp.433-440
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• 2009

High levels of inflammatory cytokines were proposed contributors to the pathogenesis of a various inflammatory skin disorders. Therefore, investigating the immune response of the inflammatory skin disorder allows a better understanding of pathogenesis of a various inflammatory skin disorders and therapeutic approaches. The aim of this study was to analyze of the immune response in dogs with chronic inflammatory skin disease. To this aim, the present study evaluated relative mRNA expression of canine $IFN-{\gamma}$, IL-4, $TGF-{\beta}$ and IL-10 using TaqMan realtime PCR assays and semi-quantitative RT-PCR in freshly isolated peripheral blood mononuclear cells from twenty dogs with chronic inflammatory skin disease and ten normal dogs. The relative mRNA expression levels of IL-4 mRNA were significantly higher in dogs with chronic inflammatory skin disease than those in normal dogs (P < 0.01). The results of present study also showed a tendency towards increased expression of IL-10 transcripts in dogs with chronic inflammatory skin disease. However, there were no significant differences in the levels $IFN-{\gamma},\;TGF-{\beta}$ between normal and chronically inflammed dogs. In addition, the concentration of serum IgE was significantly increased in dogs with chronic inflammatory skin disease compared with those in normal dogs (P < 0.01). In histopathological examination, we found that there were markedly increased mast cell counts in chronically inflammed dogs (P < 0.05). These results suggest that the pathogenesis of chronic inflammatory skin disease might be associated with a T-cell mediated inflammatory responses characterized by a Th2-skewed immune response. Based on these results, the modulation of Th1/Th2 balance may be an effective therapeutic strategy for the treatment of chronic inflammatory skin disease.

• 생명과학회지
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• 제22권8호
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• pp.1107-1113
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• 2012

동양에서 한약 재료로 사용되는 칡(Pueraria thunbergiana)은 항산화, 항균효과 및 골다공증 치료 등의 다양한 효과가 있는 것으로 밝혀지고 있지만, 생태학적으로는 산림생태계를 파괴하는 종으로 알려져 있다. 본 연구는 칡의 면역학적 효과를 검증하여 유용한 자원으로 활용하는데 목적이 있다. 칡 추출물을 이용하여 생쥐 비장에 있는 면역세포 활성화 작용을 실험한 결과, 칡 추출물은 첫째, 농도 의존적으로 생쥐 비장세포의 증식 유도하였으며 IL-6, TNF-${\alpha}$, IL-2, IFN-${\gamma}$의 cytokine 생산을 증가시켰다. 둘째, 특히 비장세포 중 칡 추출물은 B세포를 자극하여 세포증식 및 IgM의 생산을 증가시켰다. 셋째, 대식세포주의 일산화질소 생산을 유도하였으며, 또 TNF-${\alpha}$, IL-6 및 IL-$1{\beta}$의 cytokine 분비를 유도하였다. 이상의 실험 결과, 본 실험에서 사용한 칡 추출물은 B세포와 대식세포 같은 면역세포의 증식과 각종 사이토카인을 생산을 유도하기 때문에, 면역반응을 조절하는 성분이 포함되어 있는 것으로 생각되며 특히, 아세톤 추출물이 물 추출물에 비하여 효과적이었다. 따라서 추가적인 실험을 통해, 면역반응을 조절하는 성분을 분리 정제하여 그 특성을 명확히 규명한다면, 각종 의약품이나 건강식품을 개발할 수 있는 원재료로서 칡을 이용할 수 있으며, 부수적으로 산림생태계에 복원에도 기여할 수 있을 것으로 사료된다.

• 생명과학회지
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• 제24권1호
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• pp.61-66
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• 2014

본 연구는 면역억제 마우스에서 양강 에탄올 추출물이 면역증강에 미치는 영향을 평가해보았다. 마우스에 cyclophosphamide를 2회 복강 주사한 후 양강 추출물을 30, 100, 300 mg/kg 용량으로 4주간 경구투여 한 후, 체중 및 면역장기 무게, 비장세포수, 혈청 사이토카인 농도 및 혈청 면역글로불린의 농도를 측정하였다. 실험 결과 체중과 비장세포수는 양강 추출물 투여 시 대조군과 비교하여 유의한 차이를 보이지 않았다. 혈청의 IL-2, TGF-${\beta}$ 및 IFN-${\gamma}$ 농도는 AO 100군에서 대조군에 비해 유의적인 증가를 보였고(p<0.05) IL-4 농도는 실험군에서 유의한 차이를 보이지 않았다. 혈청 내 IgM의 농도는 대조군에 비해 양강 추출물 투여군 모두에서 유의적으로 증가하였고 (p<0.05), IgA의 농도는 양강 추출물 투여군에서 증가하는 경향을 보였는데 특히 AO100군에서 유의성 있게 증가하였다(p<0.05). 본 연구 결과는 양강 에탄올 추출물은 혈청 내 사이토카인 농도와 면역글로불린 농도를 증가시켜 면역력 증강에 기여할 것으로 보이며 특히 100 mg/kg을 투여하였을 때 효과가 가장 큰 것으로 나타났다.