• 대한한방부인과학회지
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• 제25권2호
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• pp.1-11
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• 2012

Objectives: This study was conducted to evaluate the inhibitory effect of Melia Fructus extract on osteoclast differentiation. Methods: MTT-assay was performed to estimate cytotoxicity of Melia Fructus extract in BMMs stimulated with M-CSF. TRAP staining, TRAP activity and Real-time PCR were performed to know the inhibitory effect on osteoclast differentiation. Actin ring formation were analysed to observe the effect of Melia Fructus extract. Results: Melia Fructus extract decreased the number of TRAP positive cells and the expression of NFATc1 gene, c-Fos gene, TRAP and OSCAR in BMMs stimulated with RANKL. Melia Fructus extract has no cytotoxicity at the concentration used in this study. Melia Fructus extract restrained the formation of actin ring. Melia Fructus inhibited NF-${\kappa}B$ activity by inducing degradation of p-$IkB{\alpha}$. Conclusions: Melia Fructus has the inhibitory effect of osteocalst differentiation and bone resorption. Further studies are needed to treat osteoporosis by herbal medicine containing Melia Fructus.

• 대한의생명과학회지
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• 제29권4호
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• pp.211-219
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• 2023

Caffeic acid phenethyl ester (CAPE) is an active component of propolis obtained from honeybee hives. CAPE possesses anti-mitogenic, anti-carcinogenic, anti-inflammatory, and immunomodulatory activities in diverse systems, which know as displays antioxidant activity and inhibits lipoxygenase activities, protein tyrosine kinase, and nuclear factor kappa B (NF-κB) activation. This study aimed to investigate the effect of CAPE on lipopolysaccharide (LPS)-induced human neutrophil phagocytosis. Human neutrophils were cultured with various concentrations of CAPE (1, 10, and 100 µM) with or without LPS. The pro-inflammatory proteins (tumor necrosis factor-alpha [TNF-α], interleukin [IL]-6 and IL-8) levels were measured after 4 h incubation. To investigate the intracellular signaling pathway, we measured the levels of mitogen-activated protein kinases (MAPK), including phosphorylation of p38, extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) and c-Jun N-terminal kinase (JNK). Next, to evaluate the potential phagocytosis, neutrophils were labeled with iron particles of superparamagnetic iron oxide nanoparticles (SPIONs, 40 nm) for 1 h in culture medium containing 5 mg/mL of iron. The labeling efficiency was determined by Prussian blue staining for intracellular iron and 3T-wighted magnetic resonance imaging. CAPE decreased the activation of intracellular signaling pathways, including ERK1/2 and c-Jun, and expression of pro-inflammatory cytokines, including TNF-α and IL-6, but had no effect on the signaling pathways of p38 and cytokine IL-8. Furthermore, images obtained after mannan-coated SPION treatment suggested that CAPE induced significantly higher signal intensities than the control or LPS group. Together, these results suggest that CAPE regulates LPS-mediated activation of human neutrophils to reduce phagocytosis.

• 한국미생물·생명공학회지
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• 제42권3호
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• pp.275-284
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• 2014

본 연구에서는 식물자원으로부터 생리활성을 보유한 새로운 기능성 소재를 찾고자 하였다. 이를 위해 수종의 중국 자생 식물을 분석하여 강한 항산화능을 보유한 3종(Malus hupehensis, Ophiorrhiza cantonensis, 그리고 Psychotria rubra)을 선별하고, 각 추출물의 항산화능과 항염증 생리활성을 분석하였다. 먼저 각 추출물의 항산화능을 DPPH radical 소거능을 통해 분석한 결과 모두 양성 대조군으로 사용한 ascorbic acid와 유사한 정도의 높은 활성을 보여 각 소재가 매우 강한 항산화능을 보유함을 확인하였다. 또한 RAW 264.7 세포주에서 $H_2O_2$ 및 LPS에 의해 유도된 ROS에 대한 각 추출물의 소거능을 분석한 결과에서도 농도의존적인 강한 소거능을 보였다. 뿐만 아니라 대표적인 항산화효소 중 하나로 항산화능 보유 천연물에 의해 발현이 유도되는 HO-1 및 그 전사 인자인 Nrf-2의 단백질 발현이 각 추출물의 처리에 의해 증가됨을 보였다. 한편 각 소재가 LPS에 의해 유도된 NO 생성에 미치는 영향을 분석한 결과 모두 농도의존적인 NO 생성 저해능을 보였으며 이는 NO 생성 단백질인 iNOS의 발현 저해에서 기인함을 확인하였다. 이와 같은 각 소재의 NO 생성 및 iNOS 발현 억제 효과는 염증 상위신호 전달계인 NF-${\kappa}B$ 및 AP-1의 조절을 통해 일어날 가능성을 보였다. 이러한 결과를 통해 중국 자생 식물 3종의 높은 항산화능과 항염증 활성을 처음으로 확인하였으며 향후 기능성 소재로서 유용하게 활용될 수 있을 것으로 판단된다.

• 생명과학회지
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• 제29권8호
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• pp.854-860
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• 2019

본 연구에서는 탈지된 갈색거저리 유충으로부터 효율적인 단백질 추출법 확립을 위한 최적 pH 조건을 조사하였다. 단백질 추출 pH 조건은 7부터 12까지 사용하였으며, 단백질 침전 pH는 2, 4, 그리고 6을 사용하였다. 이러한 조건에서 추출한 단백질의 함량, 갈변도 및 회수량을 분석한 결과 갈색거저리 유충 단백질의 최적 추출 조건은 단백질의 갈변도를 최소화 할 수 있으며 32.4%의 회수율을 나타내는 용출 pH 9.0와 침전 pH 4.0로 결정하였다. 다음으로 알칼리 추출법으로 추출한 갈색거저리 유충 단백질의 항염증 효능을 검토하였다. 갈색거저리 유충 단백질이 LPS에 의해 활성화 되는 대식세포의 염증반응 개선효과를 확인하기 위하여 염증매개 인자인 nitric oxide(NO)의 생성 및 iNOS와 COX-2 단백질의 발현량을 측정하였다. 그 결과 갈색거저리 유충 단백질은 세포독성 없이 NO의 생성을 억제 하였으며, MAPK 와 $NF-{\kappa}B$ 신호전달 억제를 통하여 iNOS와 COX-2 단백질의 발현량을 감소시켰다. 이러한 연구결과는 갈색거저리 유충 단백질이 항염증 치료제 및 단백질 보충제 개발을 위한 잠재적 가능성을 가지고 있는 것으로 사료된다.

• 대한한의학회지
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• 제44권2호
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• pp.119-131
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• 2023

Objectives: Betula Platyphylla(BP) has been used as a analgesic, anti-microbial, anti-oxidant drug in Eastern Asia. However, it is still unknown whether BP ethanol extract could exhibit the inhibitory activities against ultraviolet B(UVB)-induced skin injury on human keratinocytes, HaCaT cells. This study was aimed to investigate the protective activity of BP ethanol extract on UVB-irradiated skin injury in HaCaT cells. Methods: The skin injury model of HaCaT cells was established under UVB stimulation. HaCaT keratinocyte cells were pre-treated with BP ethanol extract for 1 h, and then stimulated with UVB. Then, the cells were harvested to measure the cell viability, production of reactive oxygen species(ROS), pro-inflammatory cytokines such as interleukin(IL) 1-beta, IL-6, and tumor necrosis factor(TNF)-𝛼, hyaluronidase, type 1 collagen, matrix metalloproteinase(MMP)s. In addition, we examined the mitogen activated protein kinases(MAPKs) and inhibitory kappa B alpha(I𝜅;-B𝛼) as inhibitory mechanisms of BP ethanol extract. Results: The treatment of BP ethanol extract inhibited the UVBinduced cell death and ROS production in HaCaT cells. BP ethanol extract treatment inhibited the UVB-induced increase of IL-1beta, IL-6, and TNF-𝛼. BP ethanol extract treatment inhibited the increase of hyaluronidase, MMP and decrease of collagen. BP ethanol extract treatment inhibited the activation of MAPKs and the degradation of I𝜅-B𝛼. Conclusions: Our result suggest that treatment of BP ethanol extract could inhibit the UVB-induced skin injury via deactivation of MAPKs and nuclear factor kappa B(NF-𝜅B) in HaCaT cells. This study could suggest that BP ethanol extract could be a beneficial agent to prevent skin damage or inflammation.

• Bulletin of the Korean Chemical Society
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• 제32권11호
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• pp.4049-4054
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• 2011

A new compound, kalopanaxin F (3), and 11 known compounds (1, 2, 4-12), were isolated from the stem bark of Kalopanax pictus. Their structures were elucidated on the basis of chemical and spectroscopic methods. Five of the compounds (2, 3, 5, 6, and 12) significantly inhibited $TNF{\alpha}$-induced NF-${\kappa}B$ transcriptional activity in HepG2 cells in a dose-dependent manner, with $IC_{50}$ values ranging from 6.2 to 9.1 ${\mu}M$. Furthermore, the transcriptional inhibitory function of these compounds was confirmed based on decreases in COX-2 and iNOS gene expression in HepG2 cells. Compounds 3-7, 9, and 12 significantly activated the transcriptional activity of PPARs dose-dependently, with $EC_{50}$ values ranging from 4.1-$12.7{\mu}M$. Compounds 4 and 5 exhibited $PPAR{\alpha}$, $PPAR{\gamma}$, and $PPAR{\beta}({\delta})$ transactivational activities in a dose-dependent manner, with $EC_{50}$ values of 16.0 and 17.0, 8.7 and 16.5, 26.2 and 26.3 ${\mu}M$, respectively.

• Asian Pacific Journal of Cancer Prevention
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• 제14권1호
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• pp.359-365
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• 2013

The high mobility group box 1 (HMGB1) protein is a multifunctional cytokine-like molecule that plays an important role in the pathogenesis of tumors. In this study, real-time polymerase chain reactions and Western blot assays indicated that HMGB1 transcriptional activity and protein level are increased in $Tax^+$-T cells (TaxP). To clarify the mechanisms, a series of HMGB1 deletion reporter plasmids (pHLuc1 to pHLuc6) were transfected into $Tax^-$-T cells (TaxN, Jurkat) and $Tax^+$-T cells (TaxP). We found that promoter activity in $Tax^+$-T cells to be higher than that in $Tax^-$-T cells, indicating a significant increase in pHLuc6. Bay11-7082 (NF-${\kappa}B$ inhibitor) treatment did not block the enhancing effect. Chromatin immunoprecipitation assays revealed that Tax was retained on a HMGB1 promoter fragment encompassing -1163 to -975. Bioinformatics analysis showed six characteristic cis-elements for CdxA, AP-1, AML-1a, USF, v-Myb, and C/EBP in the fragment in question. Mutation of cis-elements for C/EBP reduced significant HMGB1 promoter activity induced by Tax. These findings indicate that Tax enhances the expression of HMGB1 gene at the transcriptional level, possibly by interacting with C/EBP.

• The Korean Journal of Physiology and Pharmacology
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• 제5권5호
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• pp.443-449
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• 2001

Previously we reported that THI 52 inhibits tumor necrosis factor $(TNF)-{\alpha}$ mRNA expression in mouse peritoneal macrophages exposed to LPS plus $IFN-{\gamma}.$ In the present study, the effects of THI 52 on vascular reactivity ex vivo, and iNOS protein expression (rat lung) were investigated in LPS-treated rats. Treatment of THI 52 concentration-dependently reduced not only serum nitrite production but also the expression of iNOS protein in rat lung tissues. Thoracic aorta taken from LPS injected rat for 8 h ex vivo resulted in suppression of vasoconstrictor effects to phenylephrine (PE), which was restored by THI 52 (20 mg/kg) 30 min prior to LPS. When measured iNOS activity, treatment of THI 52 concentration-dependently reduced the enzyme activity in RAW 264.7 cells activated with LPS plus $IFN-{\gamma}.$ Likewise, iNOS activity was significantly reduced in lung tissues taken those rats that were injected THI 52 prior to LPS injection compared with LPS injection alone. These results strongly suggest that THI 52 can suppress iNOS gene expression induced by LPS, and restore the vascular contractility to PE. Thus, THI 52, a new synthetic isoquinoline alkaloid, may be beneficial in inflammatory disorders where production of NO is excessed by iNOS expression.

• Journal of Ginseng Research
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• 제41권2호
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• pp.188-194
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• 2017

Background: Fermented black ginseng (FBG) is processed ginseng by the repeated heat treatment and fermentation of raw ginseng. The protective effect and mechanism of FBG on cisplatin-induced nephrotoxicity was investigated to evaluate its therapeutic potential. Methods: The free radical scavenging activity of FBG was measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH). In addition, the protective effect against cisplatin-induced renal damage was tested in rats. FBG was orally administered every day at a dose of 150 mg/kg body weight for 10 d, and a single dose of cisplatin was administered intraperitoneally (7.5 mg/kg body weight) with 0.9% saline on the $4^{th}$ d. Results: The DPPH radical-scavenging activity of FBG ($IC_{50}=384{\mu}g/mL$) was stronger than that of raw ginseng. The improved DPPH radical-scavenging activity was mediated by the generation phenolic compounds. The decreased cell viability by cisplatin was recovered significantly after treatment with FBG in a dose-dependent manner. Then, the protective effect of FBG on cisplatin-induced oxidative renal damage was investigated in rats. The decreased creatinine clearance levels, which are a reliable marker for renal dysfunction in cisplatin-treated rats, were reduced to the normal level after the administration of FBG. Moreover, FBG showed protective effects against cisplatin-induced oxidative renal damage in rats through the inhibition of $NF-{\kappa}B/p65$, COX-2, and caspase-3 activation. Conclusion: These results collectively show that the therapeutic evidence for FBG ameliorates the nephrotoxicity via regulating oxidative stress, inflammation, and apoptosis.

• 한국자원식물학회지
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• 제31권5호
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• pp.466-477
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• 2018

본 연구는 헛개나무 잎, 줄기, 뿌리로부터 얻어진 메탄올 추출물의 항염증 효과를 구명하기 위해서 수행되었다. LPS로 염증이 유도된 RAW264.7 세포 내 부위별 추출물을 동시에 처리하여 염증 매개성 물질인 NO 생성량 분석 결과 $40{\mu}g/m{\ell}$농도에서 뿌리(94.7%)와 줄기(42.6%)에서는 효과가 있는 반면 잎에서는 효과가 없는 것으로 확인되었다. 이 중 탁월한 효과를 보인 뿌리 추출물(RE)의 항염증 효과 및 관련 분자적 기전을 확인하였다. 그 결과, 염증 반응의 주요 경로인 NF-kB 및 MAPK 신호전달경로에서 RE가 LPS로 유도된 NF-kB의 핵 이동을 억제하고, ERK, JNK, p38의 인산화를 억제함으로써 iNOS, COX-2의 발현이 감소되고, NO와 pro-inflammatory cytokine (IL-6, $IL-1{\beta}$, $TNF-{\alpha}$)의 생성이 억제됨을 확인하였다. 또한 RE는 대식세포에서 Nrf2를 활성화시켜 항염증성 단백질인 HO-1 발현을 유도하여 항염증 효과를 나타내었다. 또한, RE로부터 주요 성분을 분리한 후 NMR과 MS 기기를 이용하여 구조 동정된 27-O-protocatechuoylbetulinic acid 화합물에서도 높은 항염증 효과를 확인하였다. 이러한 연구결과는 헛개나무뿌리와 그 주요성분은 의약품 소재 및 기능성 식품 등의 기능성 소재로 활용될 수 있는 기초적인 정보를 제공할 것으로 생각된다.