• 대한약침학회지
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• 제14권2호
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• pp.15-24
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• 2011

Background: Type I allergy is involved in allergic asthma, allergic rhinitis, and atopic dermatitis which are accompanied by an acute and chronic allergic inflammatory responses. Rehmannia glutinosa is a traditional medicine in the East Asian region. This study examined whether a Rehmannia Glutinosa pharmacopuncture solution (RGPS) had anti-allergic or anti-inflammatory effects in antigen-stimulated-RBL-2H3 cells. Methods: We determined the effect of RGPS on cell viability using the 3-[4,5-dimethylthiazolyl]-2,5-diphenyltetrazolium bromide (MTT) assay. We also examined the effect of RGPS on the release of ${\beta}$-hexosaminidase and the secretion of IL-4 and TNF-${\alpha}$ using ELISA. In addition, we evaluated the effect of RGPS on the mRNA expression of various cytokines; IL-2, IL-3, IL-4, IL-5, IL-13 and TNF-${\alpha}$ using RT-PCR. Furthermore, we assessed the activation of mitogen-activated protein kinases (MAPKs) and nuclear factor (NF)-${\kappa}$B using Western blotting after RGPS treatment. Results: We found that RGPS ($10^{-4}$ to $10^{-1}$ dilution) did not cause any cytotoxicity. We observed significant inhibition of ${\beta}$-hexosaminidase release and suppression of the protein secretion of IL-4 and TNF-${\alpha}$ and mRNA expression of multiple cytokines in antigen-stimulated-RBL-2H3 cells after RGPS treatment. Additionally, RGPS suppressed not only the phosphorylation of MAPKs, but also the transcriptional activation of NF-${\kappa}$B in antigen-stimulated-RBL-2H3 cells. Conclusions: These results suggest that RGPS inhibits degranulation and expression of cytokines including IL-4 and TNF-${\alpha}$ via down-regulation of MAPKs and NF-${\kappa}$B activation in antigen-stimulated-RBL-2H3 cells. In conclusion, RGPS may have beneficial effects in the exerting anti-allergic or anti-inflammatory activities.

• International Journal of Oral Biology
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• 제42권3호
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• pp.117-121
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• 2017

We have previously shown that the specific phosphatidylinositol 3-kinase inhibitor LY294002 (LY29), and its inactive analog LY303511 (LY30), inhibit a monocyte chemoattractant protein-1 (MCP-1) expression in human umbilical vein endothelial cells; these results suggest the potential of LY30 as an anti-inflammatory drug. In this study, we determined the effects of LY30 on the production of various inflammatory cytokines in human macrophagic THP-1 cells which were stimulated with lipopolysaccharide (LPS). LY30 selectively suppressed the mRNA expression of IL-12 p40, $TNF-{\alpha}$, and MCP-1 without affecting the expression of $IL-1{\alpha}$, IL-6, and IL-8. Inhibition of the production of IL-12 and $TNF-{\alpha}$ by LY30 was also demonstrated using ELISA assays. In order to elucidate the mechanisms of the action of LY30, we examined the role played by the mitogen-activated protein kinases and the key transcription factors, AP-1 and $NF-{\kappa}B$ in LPS-stimulated THP-1 cells. The results revealed that LY30 inhibited LPS-induced activation of ERK, but not p38 or JNK. Furthermore, the AP-1 DNA binding activity was suppressed by LY30 based upon the dosage, whereas $NF-{\kappa}B$ DNA binding was not affected. These results suggest that LY30 selectively inhibits cytokine production in the LPS-stimulated macrophagic THP-1 cells by down-regulating the activation of ERK and AP-1.

• International Journal of Oral Biology
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• 제38권3호
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• pp.121-126
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• 2013

Tumor necrosis factor alpha ($TNF{\alpha}$) is a multifunctional inflammatory cytokine that regulates various cellular and biological processes. Increased levels of $TNF{\alpha}$ have been implicated in a number of human diseases including diabetes and arthritis. Sympathetic nervous system stimulation via the beta2-adrenergic receptor (${\beta}2AR$) in osteoblasts suppresses osteogenic activity. We previously reported that $TNF{\alpha}$ upregulates ${\beta}2AR$ expression in murine osteoblastic cells and that this modulation is associated with $TNF{\alpha}$ inhibition of osteoblast differentiation. In our present study, we explored whether $TNF{\alpha}$ induces ${\beta}2AR$ expression in human osteoblasts and then identified the downstream signaling pathway. Our results indicated that ${\beta}2AR$ expression was increased in Saos-2 and C2C12 cells by $TNF{\alpha}$ treatment, and that this increase was blocked by the inhibition of NF-${\kappa}B$ activation. Chromatin immunoprecipitation and luciferase reporter assay results indicated that NF-${\kappa}B$ directly binds to its cognate elements on the ${\beta}2AR$ promoter and thereby stimulates ${\beta}2AR$ expression. These findings suggest that the activation of $TNF{\alpha}$ signaling in osteoblastic cells leads to an upregulation of ${\beta}2AR$ and also that $TNF{\alpha}$ induces ${\beta}2AR$ expression in an NF-${\kappa}B$-dependent manner.

• Journal of Biomedical and Translational Research
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• 제19권4호
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• pp.92-102
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• 2018

The objectives of this study were to find out the plant to enhance immune activity among 42 kinds of foods frequently consumed by the Korean elderly consisting of 5 food groups and 5 wild plants. Each sample was assessed the immunoactive effect by measuring $NF-{\kappa}B/AP1$ gene expression, nitric oxide and cytokine production in $RAW-Blue^{TM}$ cell. Soybean sprouts of 47 plants showed the highest $NF-{\kappa}B/AP1$ gene expression at the level of $1.13{\pm}0.03$ (O.D. 650 nm) and Soritae, sweet potato, banana, apple, garlic, crown daisy, cabbage and Ailanthus altissima also had high activity of $NF-{\kappa}B/AP1$ gene in $RAW-Blue^{TM}$ cell stimulated by LPS. NO production of Ailanthus altissima was significantly higher than that of other plants and 16 plants of glutinous sorghum, black rice, Seoritae, Heuktae, sweet potato, banana, apple, garlic, mungbean sprouts, spinach, crown daisy, young pumpkin, cabbage, soybean sprouts, Actinidia arguta and Aster scaber were the next best activity. The above results selected 17 out of 47 plant samples. Moreover, soybean sprouts was significantly shown to increase $TNF-{\alpha}$ ($1,509.55{\pm}1.38pg/mL$) and $IL-1{\beta}$ ($54.56{\pm}1.08pg/mL$) cytokines in comparison with RAW-Blue cell stimulated by LPS. According to the results of in vitro evaluation, the ethanol extract of soybean sprout increased the production of immune-enhancing cytokines by proliferation of macrophages. In addition, $NF-{\kappa}B$ transcription factor activity and NO production ability were excellent, and it was selected as a material having excellent immunological activity.

• Biomolecules & Therapeutics
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• 제21권5호
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• pp.332-337
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• 2013

Microglial activation plays an important role in the development and progression of various neurological disorders such as cerebral ischemia, multiple sclerosis, and Alzheimer's disease. Thus, controlling microglial activation can serve as a promising therapeutic strategy for such brain diseases. In the present study, we showed that kalopanaxsaponin A, a triterpenoid saponin isolated from Kalopanax pictus, inhibited inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumor necrosis factor (TNF)-${\alpha}$ expression in lipopolysaccharide (LPS)-stimulated microglia, while kalopanaxsaponin A increased anti-inflammatory cytokine interleukin (IL)-10 expression. Subsequent mechanistic studies revealed that kalopanaxsaponin A inhibited LPS-induced DNA binding activities of NF-${\kappa}B$ and AP-1, and the phosphorylation of JNK without affecting other MAP kinases. Furthermore, kalopanaxsaponin A inhibited the intracellular ROS production with upregulation of anti-inflammatory hemeoxygenase-1 (HO-1) expression. Based on the previous reports that JNK pathway is largely involved in iNOS and proinflammatory cytokine gene expression via modulating NF-${\kappa}B$/AP-1 and ROS, our data collectively suggest that inhibition of JNK pathway plays a key role in anti-inflammatory effects of kalopanaxsaponin A in LPS-stimulated microglia.

• 한방안이비인후피부과학회지
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• 제25권1호
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• pp.12-21
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• 2012

Background and Objectives : Allergic rhinitis is one of the most common diseases in the otorhinolaryngology area. in oriental clinic, Hyunggaeyungyo-tang(HYT) has been used as a primary prescription to treat allergic rhinitis. However, there have been no studies so far performed on the effect of this HYT use. The purpose of this study was find out therapeutic effects of its exclusive use on the rat with allergic rhinitis. Material and Methods : Thirty BALB/c mice were divided into three group : normal group(NOR), control group(CON) inoculated with allergic rhinitis and sample group(SAM) treated with the HYT extract after it was treated the same as the control group. Rats were sensitized intraperitoneally with ovalbumin solution 4times at intervals of 2 days. After that time, rats in SAM were administered by HYT to treat the inflammation. Results : 1. The number of eosinophil in SAM noticeably decreased than CON and this decrease had probability. The inhibition of eosinophil distribution. The infiltration of eosinophil in SAM noticeably decreased than CON. 2. The damaged mucosa as disruption of cilia in respiratory cell, vacant mucose secreting cell and infiltration of inflammation intricate cells in CON were increased than NOR, but SAM same as normal configuration. Decrease of icthing and sneezing intricate neurotransmitter (substance P). Decrease of angiogenesis intricate cytokine(MIP-2). 3. Transcription factor(NF-${\kappa}B$ p65) was decreased. 4. Transcription factor inhibitor(p-$I{\kappa}B$) was decreased. 5. Inflammation cytokine(iNOS) was decreased. Conclusion : The results suggest that HYT is significantly effective in the treatment of inflammation caused by allergic rhinitis through the suppression of NF-${\kappa}B$ activation and iNOS production.

• 대한본초학회지
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• 제29권3호
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• pp.79-85
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• 2014

Objectives : In this study, we attempted to evaluate the effects of Careswell on human mast cell-mediated allergy inflammation in vitro and pruritogen-induced scratching behavior in vivo. Method : The Careswell was extract by distilled water. The anti-itching effects of Careswell were investigated on the compound 48/80 ($50{\mu}g/kg$) or histamine ($100{\mu}g/kg$) induced scratching behavior male ICR mice for 30 min by an observer blind. Terfenadine (10 mg/kg) was used as a positive control drug. The cell toxicity of Careswell was determined by 3-(4,5-dimethylthiazole-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. The regulatory effect of Careswell on interleukin (IL)-6 and tumor necrosis factor (TNF)-${\alpha}$ levels was determined by enzyme-linked immunosorbent assay (ELISA) in phorbol 12-myristate 13-acetate plus calcium ionophore A23187 (PMACI) stimulated human mast cells (HMC-1). Also, we evaluated the effect of Careswell on PMACI induced the activation of Nuclear factor-kappa B (NF-${\kappa}B$) into nucleus by Western blot analysis. Result : The results revealed that the oral administration of Careswell (200 mg/kg, p.o.) attenuated the compound 48/80 or histamine-induced scratching behavior in mice. We showed that Careswell significantly reduced the PMACI-induced the production of IL-6 (0.5-1 mg/ml) and TNF-${\alpha}$ (0.1-1 mg/ml). Additionally, Careswell significantly inhibited the activation of NF-${\kappa}B$ in PMACI-stimulated HMC-1. Conclusion : Collectively, the findings of this study provide us with a novel insight into the pharmacological actions of Careswell as a potential molecule for use in the treatment of allergic inflammation diseases.

• Nutrition Research and Practice
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• 제14권5호
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• pp.453-462
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• 2020

BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG), an oriental herbal medicine, has been known to improve liver function, and has both anti-inflammatory and antimicrobial properties. However, little is known about the immune-enhancing effects of PG and its mechanism. In this study, we aimed to investigate whether fermented PG extract (FPGE), which has increased platycodin D content, activates the immune response in a murine macrophage cell line, RAW 264.7. MATERIALS/METHODS: Cell viability was determined by Cell Counting Kit-8 assay and the nitric oxide (NO) levels were measured using Griess reagent. Cytokine messenger RNA levels of were monitored by quantitative reverse transcription polymerase chain reaction. To investigate the molecular mechanisms underlying immunomodulatory actions of FPGE in RAW 264.7 cells, we have conducted luciferase reporter gene assay and western blotting. RESULTS: We found that FPGE treatment induced macrophage cell proliferation in a dose-dependent manner. FPGE also modulated the expression of NO and pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1β, and IL-6. The activation and phosphorylation levels of nuclear factor kappa B (NF-κB) were increased by FPGE treatment. Moreover, 5-aminoimidazole-4-carboxamide ribonucleotide, an activator of AMP-activated kinase (AMPK), significantly reduced both lipopolysaccharides- and FPGE-induced NF-κB reporter gene activity. CONCLUSIONS: Taken together, our findings suggest that FPGE may be a novel immune-enhancing agent acting via AMPK-NF-κB signaling pathway.

• 한국식품과학회지
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• 제49권4호
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• pp.453-458
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• 2017

본 연구는 가시여지 잎 에탄올 추출물(ALE) 및 조다당 추출물(ALP)의 면역 활성에 관하여 비교하기 위하여, 선천 및 적응면역에서 중추적인 역할을 수행하는 큰포식세포에 ALE 및 ALP를 처리하여 세포 증식률, 산화질소 분비능, 사이토카인(종양괴사인자, IL-6, $IL-1{\beta}$) 분비능 및 기전 분석을 통한 신호전달에 관하여 관찰하였다. ALE 및 ALP를 큰포식세포에 처리하여 면역활성에 관하여 비교하였을 때, 큰포식세포 면역활성의 바이오-마커인 산화질소, 사이토카인의 분비능 및 iNOS의 세포내 발현이 ALP 처리구에서 유의적으로 증가되는 것으로 관찰되었으며, 기전분석결과, ALP의 처리는 MPAKs의 인산화 및 $NF-{\kappa}B$의 핵내 이동성을 증가시켜 면역활성을 증가시키는 것으로 관찰되었다. 따라서, ALP의 높은 면역활성은 MPAKs의 인산화 및 $NF-{\kappa}B$의 활성과 밀접한 관련이 있는 것으로 판단된다.

• 생명과학회지
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• 제32권11호
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• pp.899-907
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• 2022

Quercetin은 과일과 채소에 풍부한 플라보노이드 중의 하나로써, 항산화, 항염증, 항암, 항바이러스 활성 등 다양한 약리학적 활성을 가지고 있는 것으로 알려져 있다. 본 연구에서는 in vitro 모델에서 quercetin의 항염증 활성과 작용기전을 연구하였다. Quercetin은 LPS로 자극된 RAW264.7에서 세포 생존율에 영향 없이 NO 생산을 농도 의존적으로 저해하였고, iNOS와 COX-2 단백질의 발현을 억제하였다. 게다가, quercetin은 LPS로 유도된 p38, JNK, ERK의 인산화를 농도 의존적으로 저해하였고, NF-κB p65 단백질과 억제자인 IκBα 단백질의 인산화를 저해하였다. 이러한 결과는 quercetin의 항염증 활성이 MAPK 경로와 NF-κB를 조절함으로써 이루어진다는 것을 시사한다. Quercetin에 의해 4종류의 친 염증성 cytokine (CSF2, IL-1β, IL-6, TNF-α)의 발현 변화를 정량적 real-time PCR 방법으로 확인한 결과, 모든 cytokine 유전자의 발현이 감소됨을 확인하였다. 종합적으로, 본 연구결과는 플라보노이드 quercetin이 RAW264.7 세포에서 LPS로 유도된 염증반응을 MAPK 경로와 NF-κB경로를 통해 억제하고 친염증성 cytokine 유전자의 발현을 억제함으 로써 조절한다는 것을 제시한다.