• Journal of Ginseng Research
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• 제41권1호
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• pp.103-112
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• 2017

Background: 20(S)-Protopanaxadiol 20-O-D-glucopyranoside, also called compound K (CK), exerts antidiabetic effects that are mediated by insulin secretion through adenosine triphosphate (ATP)-sensitive potassium ($K_{ATP}$) channels in pancreatic ${\beta}$-cells. However, the antidiabetic effects of CK may be limited because of its low bioavailability. Methods: In this study, we aimed to enhance the antidiabetic activity and lower the toxicity of CK by including it with ${\beta}$-cyclodextrin (CD) (CD-CK), and to determine whether the CD-CK compound enhanced pancreatic islet recovery, compared to CK alone, in an alloxan-induced diabetic zebrafish model. Furthermore, we confirmed the toxicity of CD-CK relative to CK alone by morphological changes, mitochondrial damage, and TdT-UTP nick end labeling (TUNEL) assays, and determined the ratio between the toxic and therapeutic dose for both compounds to verify the relative safety of CK and CD-CK. Results: The CD-CK conjugate ($EC_{50}=2.158{\mu}M$) enhanced the recovery of pancreatic islets, compared to CK alone ($EC_{50}=7.221{\mu}M$), as assessed in alloxan-induced diabetic zebrafish larvae. In addition, CD-CK ($LC_{50} =20.68{\mu}M$) was less toxic than CK alone ($LC_{50}=14.24{\mu}M$). The therapeutic index of CK and CD-CK was 1.98 and 9.58, respectively. Conclusion: The CD-CK inclusion complex enhanced the recovery of damaged pancreatic islets in diabetic zebrafish. The CD-CK inclusion complex has potential as an effective antidiabetic efficacy with lower toxicity.

• Biomolecules & Therapeutics
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• 제29권6호
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• pp.685-696
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• 2021

In this study, we investigated the inhibitory effect of 5-aminolevulinic acid (ALA), a heme precursor, on inflammatory and oxidative stress activated by lipopolysaccharide (LPS) in RAW 264.7 macrophages by estimating nitric oxide (NO), prostaglandin E2 (PGE2), cytokines, and reactive oxygen species (ROS). We also evaluated the molecular mechanisms through analysis of the expression of their regulatory genes, and further evaluated the anti-inflammatory and antioxidant efficacy of ALA against LPS in the zebrafish model. Our results indicated that ALA treatment significantly attenuated the LPS-induced release of pro-inflammatory mediators including NO and PGE2, which was associated with decreased inducible NO synthase and cyclooxygenase-2 expression. ALA also inhibited the LPS-induced expression of pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6, reducing their extracellular secretion. Additionally, ALA abolished ROS generation, improved the mitochondrial mass, and enhanced the expression of heme oxygenase-1 (HO-1) and the activation of nuclear translocation of nuclear factor-E2-related factor 2 (Nrf2) in LPS-stimulated RAW 264.7 macrophages. However, zinc protoporphyrin, a specific inhibitor of HO-1, reversed the ALA-mediated inhibition of pro-inflammatory cytokines production and activation of mitochondrial function in LPS-treated RAW 264.7 macrophages. Furthermore, ALA significantly abolished the expression of LPS-induced pro-inflammatory mediators and cytokines, and showed strong protective effects against NO and ROS production in zebrafish larvae. In conclusion, our findings suggest that ALA exerts LPS-induced anti-inflammatory and antioxidant effects by upregulating the Nrf2/HO-1 signaling pathway, and that ALA can be a potential functional agent to prevent inflammatory and oxidative damage.

• 한국응용과학기술학회지
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• 제38권1호
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• pp.118-125
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• 2021

본 연구에서 사용된 와송(Orostachys japonica A. Berger)은 암 치료를 위한 민간요법으로 오랫동안 사용되어 온 약초이다. 본 연구에서는 와송(Orostachys japonica A. Berger) 에틸아세테이트 분획물의 항산화 효능 활성에 대하여 실험한 결과를 보고한다. 와송(Orostachys japonica A. Berger) 에틸아세테이트 분획물의 항산화 효능은 0.10 mg/mL 농도에서 78.54%의 DPPH 라디칼 소거활성을 보였고, 73.48%의 ABTS+ 라디칼 소거활성을 나타내어 항산화 효과가 있는 것을 확인하였다. 또한 대체실험동물 모델인 제브라피쉬를 이용한 독성실험 결과에서는 모든 농도에서 100%의 배아 생존율을 보이고 응고 또는 부화지연을 관찰할 수 없어 독성이 없는 것으로 나타났으며, UV-B조사에 의해 유도된 ROS생성은 와송 에틸아세테이트 분획물을 처리한 모든 larvae에서 전체적으로 형광강도가 감소하였다. 특히 3 ㎍/mL 농도에서 양성대조군 대비 35.7%의 감소율을 보여 효과적으로 ROS 생성을 억제하였음을 확인하였다. 위와 같은 결과로 미루어보아, 와송 에틸아세테이트 분획물은 천연 항산화제로서 화장품 분야에서 응용 가능성이 있을 것으로 기대한다.

• 생명과학회지
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• 제31권8호
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• pp.699-709
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• 2021

생활습관 개선을 통한 근육량향상은 대사 증후군의 위험을 줄이는 것으로 보고되었다. 본 연구에서는 갈조류인 고리매(Scytosiphon lomentaria) 에탄올 추출물이 골격근 성장억제조절 단백질인 마이오스타틴 신호전달을 억제하는지와 고 급식으로 유도된 비만 제브라피시의 대사 항상성에 대한 효과를 확인하였다. 고리매 에탄올 추출물(10 ㎍/ml)은 pGL3- (CAGA) 12-루시퍼라제 분석에서 마이오스타틴(1 nM/ml) 신호를 완전히 차단하였다. 또한 웨스턴 블롯 분석에서 마이오스타틴 신호를 차단하여 Smad2 인산화가 억제되는 것을 확인하였다. 제브라피쉬의 치어에 대한 연구는 고 급식 대조군 그룹의 체내 포도당 농도는 정상 급식 대조군 그룹보다 유의하게 높았지만, 12.5 ug의 고리매 에탄올을 처리한 고 급식 그룹과 18.75 ug의 고리매 에탄올로 처리한 고 급식 그룹의 체내 포도당 수준은 정상 급식 대조군 그룹과 유사하였다. 고 급식 그룹의 GLUT2 유전자의 mRNA 발현 수준은 정상 급식 대조군 그룹에 비해 현저히 낮았다. 하지만, 고리매 에탄올 추출물을 처리한 실험군 그룹의 GLUT2 유전자의 발현은 정상 급식 대조군 그룹의 GLUT2 유전자의 발현과 거의 유사하게 나타났다. 그러므로 고리매 에탄올 추출물은 GLUT2유전자의 발현 조절을 통하여 체내 포도당 조절이 가능함을 보여준다. 본 연구의 결과는 고리매 에탄올 추출물이 대사 증후군 치료에 도움을 주는 소재 및 마이오스타틴 억제제로서의 가능성을 시사합니다.

• Biomolecules & Therapeutics
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• 제26권2호
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• pp.146-156
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• 2018

Spermidine is a naturally occurring polyamine compound that has recently emerged with anti-aging properties and suppresses inflammation and oxidation. However, its mechanisms of action on anti-inflammatory and antioxidant effects have not been fully elucidated. In this study, the potential of spermidine for reducing pro-inflammatory and oxidative effects in lipopolysaccharide (LPS)-stimulated macrophages and zebrafish was explored. Our data indicate that spermidine significantly inhibited the production of pro-inflammatory mediators such as nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$), and cytokines including tumor necrosis $factor-{\alpha}$ and $interleukin-1{\beta}$ in RAW 264.7 macrophages without any significant cytotoxicity. The protective effects of spermidine accompanied by a marked suppression in their regulatory gene expression at the transcription levels. Spermidine also attenuated the nuclear translocation of $NF-{\kappa}B$ p65 subunit and reduced LPS-induced intracellular accumulation of reactive oxygen species (ROS) in RAW 264.7 macrophages. Moreover, spermidine prevented the LPS-induced NO production and ROS accumulation in zebrafish larvae and was found to be associated with a diminished recruitment of neutrophils and macrophages. Although more work is needed to fully understand the critical role of spermidine on the inhibition of inflammation-associated migration of immune cells, our findings clearly demonstrate that spermidine may be a potential therapeutic intervention for the treatment of inflammatory and oxidative disorders.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.477-483
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• 2023

잇꽃을 80% MeOH 수용액으로 추출하고, 얻어진 추출물을 EtOAc, n-BuOH 및 물로 용매 분획 하였다. 이 중 n-BuOH 분획으로부터 silica gel (SiO₂), octadecyl silica gel (ODS) column chromatography, 및 Prep-LC로 정제하여 4종의 화합물을 분리하였다. Nuclear magnetic resornance, mass spectroscopy 및 infrarad spectroscopy 등의 스펙트럼 데이터를 통해 화합물의 화학구조를 astragalin (1), isoquercetin (2), nicotiflorin (3), 그리고 rutin (4) 로 동정하였다. 본 연구를 통해 잇꽃 추출물, 용매 분획물 및 모든 분리 화합물의 alloxan에 의해 손상된 zebrafish 유충 췌도 보호 효과를 평가하기 위한 실험을 수행하였다. EtOAc (CTE), n-BuOH (CTB) 및 H₂O (CTW) 분획 모두 통계 유의적으로 우수한 항당뇨 효능을 확인하였으며, 분리된 화합물 1-4로 처리된 췌도 크기는 알록산 유도군 대비 각각 87.0, 88.5, 88.7, 및 89.3% 로 유의적으로 증가했다. 분리된 4종의 화합물 중 nicotiflorin은 6.752 mg/추출물(g)으로 높은 함량을 나타내었다. 이 결과를 통해 잇꽃 유래 화합물은 항당뇨 소재의 기능 지표성분으로서 활용 가능성을 확인하였다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2022년도 추계학술대회
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• pp.31-31
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• 2022

Plant-derived antioxidants are used as a healthy diet and are known to inhibit various human diseases. In this study, we investigated free radical scavenging and antioxidant activity of extracts from three plants (Ammannia multiflora, Ammannia coccinea and Salix gracilistyla) with the most DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity from 196 plant extracts inhabiting along Nakdong River in Republic of Korea. The three extracts also have strong total antioxidant activity. Moreover, the extracts inhibited hydrogen peroxide (H₂O₂)-induced reactive oxygen species production and depolarized mitochondrial membrane potential in RAW264.7 macrophages. In zebrafish larvae, 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) fluorescent intensity, induced by H₂O₂, was markedly reduced by the extracts of A. multiflora, A. coccinea and S. gracilistyla. Meanwhile, the extracts were upregulated Nrf2 and HO-1 expression, and an HO-1 inhibitor reversed the extract-induced oxidative responses both in vivo and in vitro. The data suggest that the extracts of A. multiflora, A. coccinea, and S. gracilistyla exert potential free radical scavenging and antioxidant capacities both in vivo and in vitro by activating the Nrf2/HO-1 signaling pathway.

• Nutrition Research and Practice
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• 제15권6호
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• pp.686-702
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• 2021

BACKGROUND/OBJECTIVES: Schisandrae Fructus, the fruit of Schisandra chinensis Baill., has traditionally been used as a medicinal herb for the treatment of various diseases, and has proven its various pharmacological effects, including anti-inflammatory and antioxidant activities. In this study, we investigated the inhibitory effect of Schisandrae Fructus ethanol extract (SF) on inflammatory and oxidative stress in particulate matter 2.5 (PM2.5)-treated RAW 264.7 macrophages. MATERIALS/METHODS: To investigate the anti-inflammatory and antioxidant effects of SF in PM2.5-stimulated RAW 264.7 cells, the levels of pro-inflammatory mediator such as nitric oxide (NO) and prostaglandin E₂ (PGE₂), cytokines including interleukin (IL)-6 and IL-1β, and reactive oxygen species (ROS) were measured. To elucidate the mechanism underlying the effect of SF, the expression of genes involved in the generation of inflammatory factors was also investigated. We further evaluated the anti-inflammatory and antioxidant efficacy of SF against PM2.5 in the zebrafish model. RESULTS: The results indicated that SF treatment significantly inhibited the PM2.5-induced release of NO and PGE₂, which was associated with decreased inducible NO synthase and cyclooxygenase-2 expression. SF also attenuated the PM2.5-induced expression of IL-6 and IL-1β, reducing their extracellular secretion. Moreover, SF suppressed the PM2.5-mediated translocation of nuclear factor-kappa B (NF-κB) from the cytosol into nuclei and the degradation of inhibitor IκB-α, indicating that SF exhibited anti-inflammatory effects by inhibiting the NF-κB signaling pathway. In addition, SF abolished PM2.5-induced generation of ROS, similar to the pretreatment of a ROS scavenger, but not by an inhibitor of NF-κB activity. Furthermore, SF showed strong protective effects against NO and ROS production in PM2.5-treated zebrafish larvae. CONCLUSIONS: Our findings suggest that SF exerts anti-inflammatory and antioxidant effects against PM2.5 through ROS-dependent down-regulating the NF-κB signaling pathway, and that SF can be a potential functional substance to prevent PM2.5-mediated inflammatory and oxidative damage.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 추계학술대회
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• pp.92-92
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• 2019

Excessive or chronic inflammation contributes to the pathogenesis of many inflammatory diseases such as sepsis, rheumatoid arthritis, and ulcerative colitis. Hibiscus syriacus L. has been used as a medicinal plant in many Asian countries, even though its anti-inflammatory activity has been unclear. Therefore, we investigated the anti-inflammatory effect of anthocyanin fractions from the H. syriacus L. varieties Pulsae (PS) on the lipopolysaccharide (LPS)-induced expression of proinflammatory mediators and cytokines in RAW264.7 macrophages. PS suppressed LPS-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) secretion concomitant with downregulation of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. Furthermore, PS inhibited the production of proinflammatory cytokines such as tumor necrosis factor-alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), and IL-12 in LPS-stimulated RAW264.7 macrophages. Further study showed that PS significantly decreased LPS-induced nuclear translocation of the nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) subunits, p65 and p50. Molecular docking data showed that many anthocyanins from PS fit into the hydrophobic pocket of MD2 and bound to Toll-like receptor 4 (TLR4), indicating that PS inhibits the TLR4-MD2-mediated inflammatory signaling pathway. Especially, apigenin-7-O-glucoside most powerfully bound to MD2 and TLR4 through LYS122, LYS122, and SER127 at a distance of $2.205{\AA}$, $3.098{\AA}$, and $2.844{\AA}$ and SER441 at a distance of $2.873{\AA}$ (docking score: -8.4) through hydrogen bonding, respectively. Additionally, PS inhibited LPS-induced TLR4 dimerization/expression on the cell surface, which consequently decreased MyD88 recruitment and IRAK4 phosphorylation. PS completely blocked LPS-mediated mortality in zebrafish larvae by diminishing the recruitment of neutrophil and macrophages accompanied by low levels of proinflammatory cytokines. Taken together, our results indicate that PS attenuates LPS-mediated inflammation in both in vitro and in vivo by blocking the TLR4/MD2-MyD88/IRAK4-$NF-{\kappa}B$ axis. Therefore, PS might be used as a novel modulatory candidate for effective treatment of LPS-mediated inflammatory diseases.