• 대한약침학회지
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• 제18권2호
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• pp.26-32
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• 2015

Objectives: Rheum undulatum L. has traditionally been used for the treatment of many diseases in Asia. However, its anti-proliferative activity in cancer has still not been studied. In the present study, we investigated the anti-cancer effects of methanol extract of Rheum undulatum L. (MERL) on human adenocarcinoma gastric cell lines (AGS). Methods: To investigate the anti-cancer effect of MERL on AGS cells, we treated the AGS cells with varying concentrations of MERL and performed 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assays. Cell cycle analyses, measurements of the mitochondrial membrane potential (MMP), caspase activity assays and Western blots were conducted to determine whether AGS cell death occurred by apoptosis. Results: Treatment with MERL significantly inhibited growth of AGS cells in a concentration dependent manner. MERL treatment in AGS cells leaded to increased accumulation of apoptotic sub G1 phase cells in a concentration dependent manner. In control cultures, 5.38% of the cells were in the sub G1 phase. In MERL treated cells, however, this percentage was significantly increased (9.95% at $70{\mu}g/mL$, 15.94% at $140{\mu}g/mL$, 26.56% at $210{\mu}g/mL$ and 38.08% at $280{\mu}g/mL$). MERL treatment induced the decreased expression of pro-caspase-8 and -9 in a concentration dependent manner, whereas the expression of the active form of caspase-3 was increased. A subsequent Western blot analysis revealed increased cleaved levels of poly (ADP-ribose) polymerase (PARP) protein. Also, treatment with MERL increased the activities of caspase-3 and -9 compared with the control. MERL treatment increased the levels of the pro-apoptotic truncated Bid (tBid) and Bcl2 Antagonist X (Bax) proteins and decreased the levels of the anti-apoptotic B-cell lymphoma 2 (Bcl-2) protein, whose is the stabilization of mitochondria. However, inhibitions of p38, extracellular signal regulated kinases (ERKs) and C-Jun N-terminal kinases (JNK) by MERL treatment did not affect cell death. Conclusion: These results suggest that MERL mediated cell death is associated with an intrinsic apoptotic pathway in AGS cells.

• Nutrition Research and Practice
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• 제11권1호
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• pp.3-10
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• 2017

BACKGROUND/OBJECTIVES: Sargassum horneri is an edible brown alga that grows in the subtidal zone as an annual species along the coasts of South Korea, China, and Japan. Recently, an extreme amount of S. horneri moved into the coasts of Jeju Island from the east coast of China, which made huge economic and environmental loss to the Jeju Island. Thus, utilization of this biomass becomes a big issue with the local authorities. Therefore, the present study was performed to evaluate the anti-inflammatory potential of crude polysaccharides (CPs) extracted from S. horneri China strain in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. MATERIALS/METHODS: CPs were precipitated from S. horneri digests prepared by enzyme assistant extraction using four food-grade enzymes (AMG, Celluclast, Viscozyme, and Alcalase). The production levels of nitric oxide (NO) and pro-inflammatory cytokines, including tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-$1{\beta}$ were measured by Griess assay and enzyme-linked immunosorbent assay, respectively. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), nuclear factor (NF)-${\kappa}B$, and mitogen-activated protein kinases (MAPKs) were measured by using western blot. The IR spectrums of the CPs were recorded using a fourier transform infrared spectroscopy (FT-IR) spectrometer. RESULTS: The polysaccharides from the Celluclast enzyme digest (CCP) showed the highest inhibition of NO production in LPS-stimulated RAW 264.7 cells ($IC_{50}$ value: $95.7{\mu}g/mL$). Also, CCP dose-dependently down-regulated the protein expression levels of iNOS and COX-2 as well as the production of inflammatory cytokines, including TNF-${\alpha}$ and IL-$1{\beta}$, compared to the only LPS-treated cells. In addition, CCP inhibited the activation of NF-${\kappa}B$ p50 and p65 and the phosphorylation of MAPKs, including p38 and extracellular signal-regulated kinase, in LPS-stimulated RAW 264.7 cells. Furthermore, FT-IR analysis showed that the FT-IR spectrum of CCP is similar to that of commercial fucoidan. CONCLUSIONS: Our results suggest that CCP has anti-inflammatory activities and is a potential candidate for the formulation of a functional food ingredient or/and drug to treat inflammatory diseases.

• Nutrition Research and Practice
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• 제12권3호
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• pp.199-207
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• 2018

BACKGROUND/OBJECTIVES: Fermented Laminaria japonica (FL), a type sea tangle used as a functional food ingredient, has been reported to possess cognitive improving properties that may aid in the treatment of common neurodegenerative disorders, such as dementia. MATERIALS/METHODS: We examined the effects of FL on scopolamine (Sco)- and ethanol (EtOH)-induced hippocampus-dependent memory impairment, using the Passive avoidance (PA) and Morris water maze (MWM) tests. To examine the underlying mechanisms associated with neuroprotective effects, we analyzed acetylcholine (ACh) and acetylcholinesterase (AChE) activity, brain tissue expression of muscarinic acetylcholine receptor (mAChR), cAMP response element binding protein (CREB) and extracellular signal-regulated kinases 1/2 (ERK1/2), and immunohistochemical analysis, in the hippocampus of mice, compared to current drug therapy intervention. Biochemical blood analysis was carried out to determine the effects of FL on alanine transaminase (ALT), aspartate transaminase (AST), and triglyceride (TG) and total cholesterol (TC) levels. 7 groups (n = 10) consisted of a control (CON), 3 Sco-induced dementia and 3 EtOH-induced dementia groups, with both dementia group types containing an untreated group (Sco and EtOH); a positive control, orally administered donepezil (Dpz) (4mg/kg) (Sco + Dpz and EtOH + Dpz); and an FL (50 mg/kg) treatment group (Sco + FL50 and EtOH + FL50), orally administered over the 4-week experimental period. RESULTS: FL50 significantly reduced EtOH-induced increase in AST and ALT levels. FL50 treatment reduced EtOH-impaired step-through latency time in the PA test, and Sco- and EtOH-induced dementia escape latency times in the MWM test. Moreover, anticholinergic effects of Sco and EtOH on the brain were reversed by FL50, through the attenuation of AChE activity and elevation of ACh concentration. FL50 elevated ERK1/2 protein expression and increased p-CREB (ser133) in hippocampus brain tissue, according to Western blot and immunohistochemistry analysis, respectively. CONCLUSION: Overall, these results suggest that FL may be considered an efficacious intervention for Sco- and EtOH-induced dementia, in terms of reversing cognitive impairment and neuroplastic dysfunction.

• Biomolecules & Therapeutics
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• 제20권4호
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• pp.406-412
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• 2012

This study was performed to examine the hepatoprotective effect of isorhamnetin-3-O-galactoside, a flavonoid glycoside isolated from Artemisia capillaris Thunberg (Compositae), against carbon tetrachloride ($CCl_4$)-induced hepatic injury. Mice were treated intraperitoneally with vehicle or isorhamnetin-3-O-galactoside (50, 100, and 200 mg/kg) 30 min before and 2 h after $CCl_4$ (20 ${\mu}l/kg$) injection. Serum aminotransferase activities and hepatic level of malondialdehyde were significantly higher after $CCl_4$ treatment, and these increases were attenuated by isorhamnetin-3-O-galactoside. $CCl_4$ markedly increased serum tumor necrosis factor-${\alpha}$ level, which was reduced by isorhamnetin-3-O-galactoside. The levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and heme oxygenase-1 (HO-1) protein and their mRNA expression levels were significantly increased after $CCl_4$ injection. The levels of HO-1 protein and mRNA expression levels were augmented by isorhamnetin-3-O-galactoside, while isorhamnetin-3-O-galactoside attenuated the increases in iNOS and COX-2 protein and mRNA expression levels. $CCl_4$ increased the level of phosphorylated c-Jun N-terminal kinase, extracellular signal-regulated kinase and p38, and isorhamnetin-3-O-galactoside reduced these increases. The nuclear translocation of nuclear factor kappa B (NF-${\kappa}B$), activating protein-1, and nuclear factor erythroid 2-related factor 2 (Nrf2) were significantly increased after $CCl_4$ administration. Isorhamnetin-3-O-galactoside attenuated the increases of NF-${\kappa}B$ and c-Jun nuclear translocation, while it augmented the nuclear level of Nrf2. These results suggest that isorhamnetin-3-O-galactoside ameliorates $CCl_4$-induced hepatic damage by enhancing the anti-oxidative defense system and reducing the inflammatory signaling pathways.

• 생명과학회지
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• 제26권2호
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• pp.212-219
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• 2016

다형성 신경교모세포종은 사람의 원발성 뇌종양 중 가장 흔하면서 악성이 높은 종양의 하나로 수술과 방사선치료, 화학치료 등 집중적 치료에도 불구하고 사망률이 높은 종양이다. 레스베라트롤은 다양한 자연산 물질에 포함되어 있는 폴리페놀의 일종으로 여러 종류의 암세포들에서 항암 작용이 있음이 보고되어 있으나 그 기전은 명확하게 밝혀져 있지 않다. 본 연구에서는 사람의 신경교모세포종 기원 세포인 A172 세포에서 레스베라트롤에 의한 세포 사멸 효과와 그 기전을 확인하고자 하였다. 레스베라트롤은 A172세포에서 활성산소종의 생성을 촉진하였으며 N-acetylcystein 혹은 catalase 등의 항산화제들을 전처치시 레스베라트롤의 세포 사멸 효과가 차단되었다. 레스베라트롤은 ERK와 p38 kinase, JNK 등의 인산화를 촉진하였으며 이들 인산화 효소의 억제제들을 전처치하면 레스베라트롤의 세포 사멸 효과가 차단되었다. Caspase 억제제를 전처치시 레스베라트롤에 의한 caspase-3의 활성화와 세포 사멸이 차단되었으며, N-acetylcystein을 전처치시 레스베라트롤에 의한 ERK의 활성화와 caspase-3의 활성화가 차단되었다. 이들 결과를 종합하면 레스베라트롤은 A172 세포에서 활성산소종의 생성을 촉진하며 이는 ERK와, p38 kinase, JNK 등의 활성화를 통해 caspase-의존성 기전으로 세포사멸을 유도하는 것으로 사료된다.

• 생명과학회지
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• 제23권2호
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• pp.324-331
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• 2013

O-GlcNAc 화(O-GlcNAcylation)는 단백질의 serine이나 threonine에 N-acetylglucosamine (GlcNAc) 분자가 결합하는 것으로, 기존의 당단백질과 달리 세포질 및 핵단백질 모두에 일어난다. 또한 수정의 속도가 빠르고 가역적으로 일어남이 인산화 수식과 유사하다. 그러나 수많은 인산화효소와 탈인산화효소가 관여하는 것과 달리 O-GlcNAc 수식은 O-GlcNAc transferase (OGT)와 O-GlcNAcase (OGA) 단 두 개의 효소에 의하여 이루어진다. 이러한 단순한 조절기전은 세포가 내외환경에 즉시 적응할 수 있도록 진화한 것으로 해석된다. 즉, O-GlcNAc 수식은 특정한 단백질 하나 하나의 활성을 켜거나 끄는 것이 아니라, 세포의 신호전달과정의 효율을 전반적으로 조절하는 '가변저항기(rheostat)' 역할을 한다. O-GlcNAc 수식은 흔히 같은 아미노산 혹은 그 주변의 아미노산이 인산화되는 것을 수반하는데, 이는 인산화와 함께 서로 조화를 이루어 세포활성을 조절하는 것으로 해석된다. 최근 O-GlcNAc이 더 나아가 O-GlcNAc-P로 인산화될 가능성이 제시되고 있는 바, 본 총설에서는 이의 가능성을 이론적으로 설명하고, 실제 실험결과를 소개한다.

• 농업생명과학연구
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• 제46권1호
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• pp.163-176
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• 2012

인천 연안 갯벌에서 분리한 호알카리성 Bacillus clausii I-52로부터 세포외 알카리성 단백질 분해효소(BCAP)의 발현 및 생산성을 증가시키기 위하여 BCAP promoter, ribosome 결합 서열, 신호서열, 전구체 서열 및 활성형 BCAP 유전자를 cloning한 재조합 plasmid pHPS9-fuBCAP을 penicillin-protoplast 법으로 B. clausii I-52의 염색체 DNA에 integration 하였고, 도입된 plasmid pHPS9-fuBCAP 유전자는 PCR에 의해 확인하였다. 가장 높은 단백질 분해효소 상대 활성을 보이는 선별된 transformant C5를 생산 최적화 배지(대두박 2%, 밀가루 1%, 구연산나트륨 0.5%, $K_2HPO_4$ 0.4%, $Na_2HPO_4$ 0.1%, NaCl 0.4%, $MgSO_47H_2O$ 0.01%, $FeSO_47H_2O$ 0.05%, 물엿 2.5%, 탄산나트륨 0.6%)에서 액침 배양법(배양온도, $37^{\circ}C$; 배양 시간, 48 h; 교반 속도, 650 rpm; 통기 속도, 1 vvm)으로 배양하여 단백질 분해효소를 발현 및 분비시켰을 때, BCAP 발현 양(134,670 U/ml)은 wild-type(83,960 U/ml)에 비하여 약 1.6 배 증가하였으며, 비활성도(91,611.5 U/mg 단백질)는 wild-type(71,760 U/mg 단백질)에 비하여 약 1.3 배 증가하였다. 또한, B. clausii I-52 염색체 DNA에 integration된 pHPS9-fuBCAP plasmid는 단백질 발현과 함께 8일간의 계대배양 동안에 안정하게 유지되고 있음을 확인하였다.

• 생명과학회지
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• 제29권11호
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• pp.1200-1207
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• 2019

Loganin은 Corni fructus의 주요 iridoid glycoside이며 항염증, 항당뇨 그리고 뇌신경보호 효과 등이 보고되었다. 본 연구에서는 ${\alpha}-MSH$와 IBMX처리된 B16F10세포에서 loganin의 melanogenesis억제효과의 신호전달 경로를 조사하였다. Loganin의 미백 활성을 확인하기 위해 B16F10세포에서 $1{\mu}m$에서 $20{\mu}m$사이의 농도를 처리하여 세포독성 실험을 수행한 결과 최대 $20{\mu}m$농도에서 독성을 나타내지 않았다. 또한 loganin은 ${\alpha}-MSH$와 IBMX처리된 B16F10세포에서 농도-의존적으로 멜라닌 생성을 감소시키는 것을 확인하였다. 또한 loganin의 멜라닌 생성을 억제하는 신호전달 경로를 Western blotting을 실시하여 조사하였다. Western blot결과에 따르면 loganin은 ${\alpha}-MSH$와 IBMX 처리된 B16F10세포에서 증가된 CREB인산화(Ser133)와 MITF 발현 및 tyrosinase의 유전자 발현을 감소시켰고 ERK의 인산화를 증가시켜 melanin 생성을 억제하였다. 결론적으로 loganin은 ${\alpha}-MSH$와 IBMX에 의해 유도된 과도한 멜라닌 합성을 CREB인산화와 MITF 및 tyrosinase의 유전자 발현을 억제하고 ERK의 활성화를 통해 멜라닌 합성을 감소됨을 확인하였다. 따라서 loganin은 과색소 침착과 관련된 피부질환의 보호제로서 활용될 가능성을 가지는 것으로 사료된다.

• Journal of Ginseng Research
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• 제39권1호
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• pp.29-37
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• 2015

Background: Panax ginseng (i.e., ginseng) root is extensively used in traditional oriental medicine. It is a modern pharmaceutical reagent for preventing various human diseases such as cancer. Ginsenosidesd-the major active components of ginsengd-exhibit immunomodulatory effects. However, the mechanism and function underlying such effects are not fully elucidated, especially in human monocytes and dendritic cells (DCs). Methods: We investigated the immunomodulatory effect of ginsenosides from Panax ginseng root on $CD14^+$ monocytes purified from human adult peripheral blood mononuclear cells (PBMCs) and on their differentiation into DCs that affect $CD4^+$ T cell activity. Results: After treatment with ginsenoside fractions, monocyte levels of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, and IL-10 increased through phosphorylation of extracellular signal-regulated kinase (ERK)1/2 and c-Jun N-terminal kinase (JNK), but not p38 mitogen-activated protein kinase (MAPK). After treatment with ginsenoside fractions, TNF-${\alpha}$ production and phosphorylation of ERK1/2 and JNK decreased in lipopolysaccharide (LPS)-sensitized monocytes.We confirmed that DCs derived from $CD14^+$ monocytes in the presence of ginsenoside fractions (Gin-DCs) contained decreased levels of the costimulatory molecules CD80 and CD86. The expression of these costimulatory molecules decreased in LPS-treated DCs exposed to ginsenoside fractions, compared to their expression in LPS-treated DCs in the absence of ginsenoside fractions. Furthermore, LPS-treated Gin-DCs could not induce proliferation and interferon gamma (IFN-${\gamma}$) production by $CD4^+$ T cells with the coculture of Gin-DCs with $CD4^+$ T cells. Conclusion: These results suggest that ginsenoside fractions from the ginseng root suppress cytokine production and maturation of LPS-treated DCs and downregulate $CD4^+$ T cells.

• 대한화장품학회지
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• 제31권4호
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• pp.311-321
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• 2005

차(Camellia sinensis)는 세계적으로 애용되는 음료이다. 그 중, 아시아권에서 주로 소비되는 녹차는 다양한 생리활성을 가지고 있어 화장품, 기능성식품을 위한 기능성 소재로서 선호되고 있다. 녹차의 피부에 대한 활성은 세포보호에서부터 피부 기질 단백질의 합성까지 다양하게 나타난다. 녹차 폴리페놀(green tea polyphenols; GTPs)은 활성을 나타내는 주성분으로서 항산화 활성 이외에 항암, 항염증, 피부면역능 저하방지활성 등을 보이며, 세포내 신호전달 경로에도 관여한다 GTPs는 표피각질형성세포에서 자외선 조사에 의한 산화 스트레스를 감소시키고, 그에 따르는 mitogen-activated protein kinase (MAPK) 신호전달과 세포사멸을 억제한다. 또한, 같은 세포에서 tumor necrosis factor alpha $(TNF{\alpha})$나 다른 화학물질에 의해 cyclooxygenase-2(COX-2), interleukin-8 (IL-8) 및 vascular endothelial growth factor (VEGF) 같은 염증매개물질이 유발되는 것을 막아준다. 또한, GTPs는 등물실험에서 화학물질이나 자외선에 의한 피부암의 발생도 억제하는데, 경구투여 외에 경피투여로도 효과가 있었다. 피부보호작용 이외에도 GTPs는 각질형성세포의 분화촉진, 노화된 피부세포의 증식능 회복, 피부기질단백질의 분해억제 등의 기능이 있으며, 피부세포에서의 기질단백질 생합성을 직접적으로 촉진하기도 한다. 녹차성분이 보이는 이러한 피부에 대한 활성은, 제형 측면에서의 연구가 진행되어 감에 따라 보다 효과적인 피부를 위한 소재로서의 사용 가능성을 더욱 높여주고 있다.