• 대한한의학회지
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• 제30권2호
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• pp.133-144
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• 2009

Background and Objectives: Sinusitis, referring to an inflammatory disease that occurs on the mucus membrane inside the sinus, is one of the most common diseases in the otorhinolaryngology area. In oriental clinic, Sunbanghwalmyungeum (SHE) has been used as a primary prescription to treat inflammatory diseases and intumescence and to treat sinusitis. The aim of this study was to investigate the anti-inflammatory and anti-allergic effects of SHE on acute sinusitis induced mice. Materials and Methods: BALB/c mice were divided into three groups: the normal group, the group inoculated with S. pneumoniae which caused them allergic rhinitis (control group), and the group treated with the SHE extract after it was treated the same as the control group (sample group). We investigated the inhibition of Th 2 cell differentiation by SHE and the suppression of NF-${\kappa}B$ activation. Results: NF-${\kappa}B$ activation was suppressed, and iNOS & COX-2 production were inhibited by SHE in acute sinusitis. IL-4 and STAT 6 also appeared to be suppressed. The number of eosinophils in the sample group noticeably decreased when compared to the control group. In the general morphologic change, the increase of damaged respiratory ciliated epithelium & eosinophil's infiltration were decreased in the sample group. Goblet cells were maintained in the sample group. MIP-2 and HSP-70 decreased in the sample group. Apocrine secretion decreased in the sample group. Conclusion: The results suggest that SHE is significantly effective in the treatment of inflammation caused by acute sinusitis through the suppression of NF-${\kappa}B$ activation and the inhibition of Th 2 cell differentiation.

• 한국식품영양과학회지
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• 제33권7호
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• pp.1092-1097
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• 2004

큰느타리버섯의 기능성 식품으로서 활용도를 높이기 위해 동결 건조된 자실체에서 분리한 조다당체 추출물이 면역세포 활성에 미치는 효과를 조사한 결과는 다음과 같다. 조다당체 추출물은 300 및 1,000 $\mu$g/mL 농도에서 비장세포의 증식을 유도하였으며, 이 때 비장세포는 IL-6와 IFN-${\gamma}$ 분비를 유도하는 것으로 나타났다 조다당체 추출물은 농도 의존적으로B세포의 증식을 유도하였으며, 특히 100 $\mu$g/mL농도 이상에서는B세포의 증식이 현저히 증가하는 것으로 나타났다. 그리고 조다당체 추출물 1,000 $\mu$g/mL 농도에서 B세포가 생산하는 IgGl, IgG2a, IgG3의 분비량이 현저히 증가하였다. 또한 농도 의존적으로 대식세포주의 일산화질소 생산을 유도하였으며, 대식세포가 분비하는 IL-6, TNF-$\alpha$, GM-CSF의 생산도 현저히 증가하는 것을 확인할 수 있었다.

• Molecules and Cells
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• 제22권1호
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• pp.104-112
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• 2006

We previously reported that DA-9601, ethanol herbal extract of Artemisia asiatica, inhibited histamine and leukotriene releases in guinea pig lung mast cells activated with specific antigen/antibody reaction. This study aimed to evaluate the inhibitory effect of DA-9601 on the OVA-induced airway inflammation in allergic asthma mouse model. BALB/c mice were sensitized and challenged with OVA. DA-9601 was administered orally 1 h before every local OVA-challenge. OVA-specific serum IgE was measured by ELISA, recruitment of inflammatory cells in BAL fluids and lung tissues by Diff-Quik and H&E staining, respectively, the expressions of CD40, CD40L and VCAM-1 by immunohistochemistry, goblet cell hyperplasia by PAS staining, activities of MMPs by gelatin zymography, expressions of mRNA and proteins of cytokines by RT-PCR and ELISA, activities of MAP kinases by western blot, and activity of NF-${\kappa}B$ by EMSA. DA-9601 reduced IgE level, recruitment of inflammatory cells into the BAL fluid and lung tissues, expressions of CD40, CD40L and VCAM-1 molecules, goblet cell hyperplasia, MMPs activity, expressions of mRNA and productions of various cytokines, activities of MAP kinases and NK-${\kappa}B$ increased from OVA-challenged mice. These data suggest that DA-9601 may be developed as a clinical therapeutic agent in allergic diseases due to suppressing the airway allergic inflammation via regulation of various cellular molecules expressed by MAP kinases/NF-${\kappa}B$ pathway.

• 대한본초학회지
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• 제35권5호
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• pp.23-31
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• 2020

Objectives : This study was conducted to evaluate the anti-hyperlipidemia effect of Scutellariae Radix, Aucklandiae Radix and Bupleuri Radix(SAB). Methods : FL83B cells were mouse liver hepatocytes, and we used this cell line. FL83B cells were treated with 0.5 mM oleic acid(OA) for 24 h, SAB extract was treated. After OA treatment, intracellular triglyceride (TG) and free fatty acid contents were measured with AdiopoRed™ assay and Free Fatty Acid Quantitation assay kit, respectively. Further, we evaluated several lipogenesis and metabolic markers such as sterol regulatory element-binding transcription factor-1c (SREBP-1c), acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), 3-hydroxy3-methyl-glutaryl CoA reductase (HMGCR), hormone-sensitive lipase (HSL), carnitine palmitoyltransferase (CPT-1), peroxisome proliferator activated receptor alpha (PPARα), and cluster of differentiation (CD36) using RT-PCR and Western-blot analysis. Results : OA markedly increased intracellular TG and free fatty acid, which plays a key role in reducing hepatic lipid accumulation, in FL83B cells. These increases were alleviated by SAB extract. The mRNA and protein expression of Fatty acid(FA) oxidation factors (CPT-1, PPARα), lipolysis factor(HSL), FA transporter(CD36), cholesterol synthesis factors (HMGCoA) and Lipodenesis (SREBP-1c, FAS, and ACC-1) were significantly increased by treatment of SAB extract in the OA-induced fatty liver cell model. Conclusions : In summary, the treat of SAB extract showed a significant reduction of the influx of fatty acids into hepatocytes, promoted the oxidation of fatty acids, and regulated fat synthesis-related factors, thereby regulating the accumulation of TG and free fatty acids.

• 한국약용작물학회지
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• 제8권4호
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• pp.291-296
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• 2000

구기자나무의 부위별 물추출물이 LPS (lipopolysaccharide) 및 Con A (concanavalin A)에 의한 마우스의 비장세포 증식능에 미치는 영향을 시험한 결과 부위별 면역 활성중 과실에서 의 No mil 처리구에서는 $0.1mg/ml{\sim}0.5mg/ml$ 농도에서, LPS 처리구에서는 $0.01mg/ml{\sim}0.1mg/ml$의 농도에서 B-cell (체액성 면역)의 분열증식을 촉진시켰다. 잎과 뿌리는 LPS 처리구에서 0.1mg/ml 까지 비장세포의 분열증식을 촉진시켰으며 Con A 처리구에서는 T-cell (세포성 면역)의 증식효과가 없었다.

• 한국정보통신학회:학술대회논문집
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• 한국정보통신학회 2019년도 춘계학술대회
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• pp.528-532
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• 2019

본 연구는 뉴런 세포와 슈반 세포의 공동 배양에 의한 수초화 발생 과정과 herpes simplex virus-1 감염에 의한 탈수초화 발생과정을 전자 현미경과 분자생물학적 분석에 의하여 확인하고자 하였다. 쥐의 배아로부터 후근신경절(dorsal root ganglion, DRG)을 분리하여 슈반(Schwann) 세포와 뉴런 세포(neuronal cell)를 in vitro에서 각각 배양하였다. 유사 분열 억제인자로 처리한 뉴런세포와 정제된 슈반 세포를 함께 공동 배양을 하여 수초화를 발생시켰다. 이렇게 수초화된 공동 배양 세포에 herpes simplex virus-1를 감염시켜 탈수초화를 진행시켰다. 수초 형성의 존재를 의미하는 myelin protein zero(MPZ) 항체를 사용하고 전자 현미경을 이용하여 수초 발생 및 탈수초화 과정을 관찰하였다. 이 연구는 과학 기술부, ICT 및 미래 계획 (NRF-2016R1A2B4016552 및 2017R1A2B3005753)이 자금을 지원하는 국립 연구 재단 (NRF)을 통한 기초 연구 프로그램의 지원을 받았다.

• 한방재활의학과학회지
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• 제31권4호
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• pp.1-11
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• 2021

Objectives Hwanggeumjakyak-tang (HJT) has traditionally been used to treat gastrointestinal inflammatory diseases; however, its protective effects against neuronal inflammation are still undiscovered. Methods We investigated the anti-neuroinflammatory effects of HJT water extract on lipopolysaccharide (LPS)-stimulated BV2 mouse microglia cells. BV2 cells were treated with LPS (1 ㎍/mL) 1 hour prior to the addition of HJT. We measured cell viability using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and nitrite production using the Griess assay. We performed a reverse transcription-polymerase chain reaction assay to measure messenger RNA expression of inflammatory cytokines including interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. Western blot analysis was performed to determine protein expression of mitogen-activated protein kinases (MAPKs) and inhibitor of nuclear factor kappa B (NF-κB)α. Results HJT inhibited excessive nitrite release in LPS-stimulated BV2 cells and also significantly inhibited inflammatory cytokines such as IL-1β, IL-6, and TNF-α in LPS-stimulated BV2 cells. Moreover, HJT significantly suppressed LPS-induced MAPK and NF-κB activation and inhibited the elevation of IL-1β, IL-6, and TNF-α in the brain of LPS-injected mice. Conclusions Our study highlights the anti-neuroinflammatory effects of HJT via MAPK and NF-κB deactivation.

• Molecules and Cells
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• 제46권12호
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• pp.764-777
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• 2023

Recombinant immunotoxins (RITs) are fusion proteins consisting of a targeting domain linked to a toxin, offering a highly specific therapeutic strategy for cancer treatment. In this study, we engineered and characterized RITs aimed at mesothelin, a cell surface glycoprotein overexpressed in various malignancies. Through an extensive screening of a large nanobody library, four mesothelin-specific nanobodies were selected and genetically fused to a truncated Pseudomonas exotoxin (PE24B). Various optimizations, including the incorporation of furin cleavage sites, maltose-binding protein tags, and tobacco etch virus protease cleavage sites, were implemented to improve protein expression, solubility, and purification. The RITs were successfully overexpressed in Escherichia coli, achieving high solubility and purity post-purification. In vitro cytotoxicity assays on gastric carcinoma cell lines NCI-N87 and AGS revealed that Meso(Nb2)-PE24B demonstrated the highest cytotoxic efficacy, warranting further characterization. This RIT also displayed selective binding to human and monkey mesothelins but not to mouse mesothelin. The competitive binding assays between different RIT constructs revealed significant alterations in IC₅₀ values, emphasizing the importance of nanobody specificity. Finally, a modification in the endoplasmic reticulum retention signal at the C-terminus further augmented its cytotoxic activity. Our findings offer valuable insights into the design and optimization of RITs, showcasing the potential of Meso(Nb2)-PE24B as a promising therapeutic candidate for targeted cancer treatment.

• 생명과학회지
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• 제21권8호
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• pp.1076-1082
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• 2011

미세소관(microtubule) 위를 이동하는 키네신은 분비소포를 이동시키는 운동단백질이다. KIF5s (KIF5A, KIF5B and KIF5C)는 세포막으로 싸인 각종 세포 내 소기관과 결합하여 미세소관을 따라 목적지까지 이동시킨다는 결과는 알려져 있지만, 어떻게 상대의 cargo를 인식하는지는 밝혀지지 않았다. 본 연구는 KIF5B의 결합 단백질을 동정하기 위하여 효모 two-hybrid system을 사용하여 KIF5B와 특이적으로 결합하는 Myo9b을 확인하였다. Myo9b는 액틴위를 이동하는 운동단백질로 다른 KIF5s들과도 결합함을 효모 two-hybrid assay로 확인하였다. 또한 Myo9s의 GTPase 활성화 단백질(GAP) 영역은 KIF5B와 결합하는데 필수영역임을 확인하였고, 이러한 단백질간의 결합은 Glutathione S-transferase (GST) pull-down assay를 통하여서도 확인하였다. 생쥐의 뇌 파쇄액에 KIF5B들의 항체로 면역침강을 행하여 Myo9s 단백질을 확인한 결과, KIF5s는 Myo9s 단백질과 특이적으로 함께 침강하였다. 이러한 결과들은 kinesin-I는 액틴 결합 운동단백질과 직접 결합함을 보여준다.

• 한국가금학회지
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• 제27권1호
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• pp.73-78
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• 2000

Embryonic stem (ES) cells are pluripotent cell lines, which derived from preimplantation embryo. These cells have been used as a vehicle of foreign DNA for production of transgenic mammals. this experiment was performed to examined the possible use of blastodermal cells derived from hen's egg for germline manipulation. Stage X blsdtodermal cells isolated from fertilized eggs were cultured in DMEM containing 15% fetal calf serum. Blastodermal cells wre co-cultured on the chicken embryonic fibroblast (CEF) or mouse embryonic fibroblast(MEF) cells. to examine the effects of growth factors on stem cell growth, bFGF and LIF were added. There was no significant difference in colony formation of putative ES cells between CEF and MEF as a feederlayer, but the addition of growth factors enhanced the proliferation and inhibited differentiation of blastodermal cells. To characterize the cell colonies as a putative ES cells, putative embryonic cell colonies were stained by periodic acid Schiffs (PAS) reagent. The putative ES cell colonies showed intensive positive reaction similar to the property of undifferentiated PGC upto 20days in vitro, but not in other cell types. this result demonstrates that PAS-positive cell colonies may be used for the study of establishment of chicken ES cell lines for the production of transgenic chicken.