• 한국식품과학회지
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• 제52권2호
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• pp.138-143
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• 2020

본 연구에서는 다양한 만성 대사성 질환에 기초적 유발 기전인 산화적 스트레스 및 염증에 대한 폴리페놀 함유 블랙 초크베리의 보호 효과를 살펴보았다. 블랙 초크베리는 DPPH, ·OH, 및 NO 등 다양한 라디컬 소거능을 나타냈다. 내독소인 LPS에 의해 활성화된 대식세포 내 ROS 증가는 폴리페놀 함유 블랙 초크베리에 의해 감소하였다. 이는 블랙 초크베리가 전사인자인 NRF2의 핵 내 이동 및 이로 인해 조절되는 SOD에 의한 ROS 소거능 및 NOX2에 의한 ROS 생성 저하에 의한 것으로 나타났다. 블랙 초크베리의 염증 보호 효과는 활성화된 대식세포에서 증가하는 NF-κB의 핵 내 이동에 따른 COX-2, iNOS와 같은 염증 매개인자 관련 효소와 TNF-α, IL-6, IL-1β와 같은 염증성 사이토카인의 저하에 의한 것으로 나타났다. 결론적으로 블랙 초크베리의 산화적 스트레스 및 염증 보호 효과는 항산화 효소 관련 전사 인자인 NRF2 조절 기전 및 염증 관련 전사 인자인 NF-κB와 관련 유전자 발현의 조절을 통하여 나타났다. 추후 블랙 초크베리 내 기능성 물질에 대한 심도 있는 기전 연구를 통해 천연물 유래 기능성 소재로서의 역할을 할 것으로 판단된다.

• 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.

• 생약학회지
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• 제51권4호
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• pp.244-254
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• 2020

Atractylodes macrocephala is a perennial herb and is a member of the Compositae family. This plant is known to contain various bioactive constituents indicating anti-inflammatory, neuroprotective, anti-oxidant, immunological enhancement, and gastroprotective effects. In this investigation, we isolated four compounds with similar chemical structures from A. macrocephala, and evaluated their anti-inflammatory effects. Among the four compounds, compound 2(atractylenolide II) showed the second-best inhibitory effect on the lipopolysaccharide(LPS)-induced production of nitric oxide in RAW264.7 macrophages and BV2 microglial cells. Compound 2 also inhibited the LPS-induced the production of prostaglandin E2(PGE2), and the expression of inducible nitric oxide synthase(iNOS) and cyclooxygenase(COX)-2 proteins in both cells. In addition, compound 2 suppressed the production of pro-inflammatory cytokines including interleukin(IL)-1β, IL-6, and tumor necrosis factor(TNF)-α. These inhibitory effects were contributed by inactivation of nuclear factor kappa B(NF-κB) and mitogen-activated protein kinases(MAPKs) pathways by treatment with compound 2. This compound did not induce the expression of heme oxygenase(HO)-1 protein indicating that the anti-inflammatory effect of compound 2 was independent with HO-1 protein. Taken together, these results suggested that atractylenolide II can be a candidate material to treat inflammatory diseases.

• 동의생리병리학회지
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• 제34권6호
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• pp.348-354
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• 2020

Oryeong-san (ORS) is a traditional Korean herbal medicine widely used for renal associated diseases, composed of five medicine herbs; Atractylodes japonica Koidzumi, Cinnamomum cassia Presl, Polyporus umbellatus Fries, Poria cocos Wolf and Alisma orientale Juzepzuk. We studied to improve the convenience of intake and portability by developing modernized dosage forms, and examined the effect on anti-inflammation of ORS. In order to develop the tablet formulation of ORS (ORS-F), the tablets were evaluated on the basis of physical characteristics include diameter, thickness, weight variation, hardness, friability and disintegration. To analyze the marker components of ORS-F, eight index markers from five herbal medicines were chosen. And the method using high performance liquid chromatography (HPLC) with diode-array detector method was established for the simultaneous analysis. The biological activities were examined the effect of ORS-F on pro-inflammation mediated by LPS-stimulation. The production of nitric oxide (NO) and cytokines were determined by reacting cultured medium with griess reagent and enzyme-linked immunosorbent assay (ELISA). The expression of cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS) were investigated by Western blot and RT-PCR. The anti-oxidant activities of OJS-F increased markedly, in a dose-dependent manner. and, The total phenolic compound and flavonoids contents of OJS-F were 10.20±0.09 ㎍/㎎ and 12.86±0.86 ㎍/㎎. OJS-F which is LPS has diminished in the LPS-induced release of inflammatory mediators (NO, iNOS, COX2 and PGE2) and pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) from the RAW264.7 macrophages. Therefore, the developed formulation for tablet of ORS-F provide efficiency and usability, and indicated effect of anti-inflammation.

• 대한한의학방제학회지
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• 제29권2호
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• pp.81-91
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• 2021

Objectives : This study was conducted to evaluate the efficacy of a complex mixture of natural substances of ginseng and baeknyeoncho on the arthritic rats. Methods : In vitro experiments were conducted to ensure the stability of the complex. After setting toxicity and concentration by MTT assay, the antioxidant effect was measured through DPPH and ABTS radical scavenging activity. To confirm the anti-inflammatory effects of the complex, levels of nitric oxide (NO) and pro-inflammatory cytokines (IL-1β, TNF-α) were measured in LPS-treated macrophage cell lines (RAW264.7). We injected monosodium iodoacetate (MIA) 50 μl (60 mg/ml) into knee joints of rats to induce osteoarthritis. The rats were divided into three groups (normal (n=5), control (n=5), and OR (n=5) group). The control group consumed 2 mg/kg of physiological saline once a day for 4 weeks, and the OR group was mixed at a concentration of 416.5 mg/kg of Baengnyeoncho (O) and 208.25 mg/kg of red ginseng (R) and ingested 1 mL each 5 days a week. Results : This complex increased the DPPH and ABTS radical scavenging rate. The complex decreased NO production and pro-inflammatory cytokine production of macrophages. In the OR group, the secretion of cytokine in serum was decreased. In histopathological examination, the joint tissue of the composite showed less damage to the synovial membrane, cartilage, and fibrous tissue than the control group. Conclusions : As a result of this study, natural complexes have antioxidant, anti-inflammatory and cartilage protection effects. Therefore, we expect the complex to be effective in treating osteoarthritis.

• Journal of Microbiology and Biotechnology
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• 제32권9호
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• pp.1154-1167
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• 2022

In this study, we investigated the anti-amnesic effect of Korean red pine (Pinus densiflora) bark extract (KRPBE) against amyloid beta_1-42 (Aβ_1-42)-induced neurotoxicity. We found that treatment with KRPBE improved the behavioral function in Aβ-induced mice, and also boosted the antioxidant system in mice by decreasing malondialdehyde (MDA) content, increasing superoxide dismutase (SOD) activities, and reducing glutathione (GSH) levels. In addition, KRPBE improved the cholinergic system by suppressing reduced acetylcholine (ACh) content while also activating acetylcholinesterase (AChE), regulating the expression of choline acetyltransferase (ChAT), postsynaptic density protein-95 (PSD-95), and synaptophysin. KRPBE also showed an ameliorating effect on cerebral mitochondrial deficit by regulating reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and ATP levels. Moreover, KRPBE modulated the expression levels of neurotoxicity indicators Aβ and phosphorylated tau (p-tau) and inflammatory cytokines TNF-α, p-IκB-α, and IL-1β. Furthermore, we found that KRPBE improved the expression levels of neuronal apoptosis-related markers BAX and BCl-2 and increased the expression levels of BDNF and p-CREB. Therefore, this study suggests that KRPBE treatment has an anti-amnestic effect by modulating cholinergic system dysfunction and neuroinflammation in Aβ_1-42-induced cognitive impairment in mice.

• Nutrition Research and Practice
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• 제16권6호
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• pp.685-699
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• 2022

BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG) has long been known as a medicinal herb effective in various diseases, including bronchitis and asthma, but is still more widely used for food. Fermentation methods are being applied to increase the pharmacological composition of PG extracts and commercialize them with high added value. This study examines the hydrolyzed and fermented PG extract (HFPGE) fermented with Lactobacillus casei in RAW 264.7 cells, and investigates the effect of amplifying the immune and the probable molecular mechanism. MATERIALS/METHODS: HFPGE's total phenolic, flavonoid, saponin, and platycodin D contents were analyzed by colorimetric analysis or high-performance liquid chromatography. Cell viability was measured by the MTT assay. Phagocytic activity was analyzed by a phagocytosis assay kit, nitric oxide (NO) production by a Griess reagent system, and cytokines by enzyme-linked immunosorbent assay kits. The mRNA expressions of inducible nitric oxide synthase (iNOS) and cytokines were analyzed by reverse transcription-polymerase chain reaction, whereas MAPK and nuclear factor (NF)-κB activation were analyzed by Western blots. RESULTS: Compared to PGE, HFPGE was determined to contain 13.76 times and 6.69 times higher contents of crude saponin and platycodin D, respectively. HFPGE promoted cell proliferation and phagocytosis in RAW 264.7 cells and regulated the NO production and iNOS expression. Treatment with HFPGE also resulted in increased production of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, C-X-C motif chemokine ligand10, granulocyte-colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and monocyte chemoattractant protein-1, and the mRNA expressions of these cytokines. HFPGE also resulted in significantly increasing the phosphorylation of NF-κB p65, extracellular signal-regulated kinase, and c-Jun N-terminal kinase. CONCLUSIONS: Taken together, our results imply that fermentation and hydrolysis result in the extraction of more active ingredients of PG. Furthermore, we determined that HFPGE exerts immunostimulatory activity via the MAPK and NF-κB signaling pathways.

• 대한본초학회지
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• 제35권3호
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• pp.17-24
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• 2020

Objectives : The objective of this study was to evaluate the gastric protective effect of Curcuma Longae Rhizoma (CLR) in 150 mM HCl/60% ethanol induced gastric ulcer (GU) in mice. Methods : Forty ICR mice were divided into five groups (n=8/Group): Nor group; Normal, Veh group; GU control, SC group; GU + sucralfate 10 mg/kg, CL; GU + CLR 30% ethanol extract 100 mg/kg, CH group; GU + CLR 30% ethanol extract 200 mg/kg. Then, mice were orally administered with 150 mM HCl/60% ethanol and caused GU. After 1 hr, mice were sacrificed, and blood and stomach tissue were collected. Results : CLR showed significance scavenging effects in 1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) radical scavenging activities (DPPH IC₅₀; 78.18 ± 0.60 ㎍/㎖, ABTS IC₅₀; 55.91 ± 1.86 ㎍/㎖). CLR significance reduce inflammatory-related factors such as cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), and interleukin-6 (IL-6) via nuclear factor kappa B (NF-κB) inactivation. In addition, the activation of nuclear factor erythroid2-related factor 2 (Nrf2) significantly led to up-regulation of anti-oxidant enzymes including factors heme oxygenase-1 (HO-1), super oxide dismutase (SOD), and glutathione peroxidase-1/2 (GPx-1/2). Conclusions : Our discovery provides that CLR possesses anti-oxidant and anti-inflammatory effects. Hence, CLR may ameliorate the development of gastric ulcer though the inhibition of NF-κB inflammatory pathway and the elevation of Nrf2 anti-oxidant pathway.

• 한국어병학회지
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• 제35권1호
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• pp.103-111
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• 2022

This study has been performed to investigate the potential effects of by-product discarded after probiotics production (BPPP) on growth performance, immune gene expression, innate-immunity status, and disease resistance of carp, Cyprinus carpio. For 3 weeks, carp were fed four diets containing different levels of BPPP at 0, 0.1, 0.2 and 0.5% per kg of normal diet. Every 7 days of feeding, immune-related gene expression, serum lysozyme activity and ACH₅₀ were analyzed. Growth rates and challenge test with E. tarda were conducted after 3 weeks of BPPP feeding. Both lysozyme activity and ACH₅₀ were significantly (p<0.05) increased in all BPPP supplemented groups compared to the control at every 7 day for 3 weeks of feeding trial. The gene expression of pro-inflammatory cytokines, IL-1β and TNF-α was significantly (p<0.05) up-regulated until 21 days of feeding in all groups except for 0.2% group on day 7 post feeding. The anti-inflammatory cytokine IL-10 gene expression was only significantly (p<0.05) increased in 0.1% group on day 7 and decreased (p<0.05) on day 14 in all BPPP supplemented groups. On day 21, the IL-10 gene expression was augmented (p<0.05) in all groups. SOD gene expression was significantly (p<0.05) increased compared to the control on day 14 and 21 post feeding, whereas no significant difference was observed on day 7. In challenging test, 0.2%, 0.1%, 0.5% and control group showed 80%, 70%, 60% and 40% of survival rate, respectively. Feed conversion rate was only improved in 0.5% group. In conclusion, the present study indicates that dietary BPPP suplementation improved growth performance, innate immune response and bactericidal activity in carp.

• 생명과학회지
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• 제32권9호
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• pp.712-720
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• 2022

비록 PM_2.5 노출과 다양한 안구 표면 질환과 관련성이 많은 선행 연구에서 알려졌지만, PM_2.5 가 각막에 미치는 세포 독성에 대한 연구는 거의 수행되지 않았다. 본 연구의 목적은 PM에 의한 각막 상피세포의 유해성을 평가하기 위한 in vitro 모델로서 쥐의 각막유래 상피세포(primary rat corneal epithelial cells, RCE cells)의 효능을 조사하는 것이다. 이를 위하여 쥐의 눈에서 분리한 1차 배양 세포가 각막 상피세포임을 pan-cytokeratin 염색을 통하여 확인하였으며, PM_2.처리에 의한 각막 상피세포의 형태학적 변화를 동반한 생존율의 억제는 세포사멸 유도와 관련이 있었다. 또한 PM_2.가 처리된 각막 상피세포에서는 ROS의 생성이 증가되었으며, 이는 미토콘드리아 기능 장애와 연관성이 있었다. 이와 함께 PM_2.는 각막 상피세포에서 NO, TNF-α, IL-1β 및 IL-6를 포함한 염증 매개인자 및 사이토카인의 생성을 증가시켰다. 아울러 heatmap 분석을 통해 BLNK, IL-1RA, Itga2b, ABCb1a 및 Ptgs2가 미세먼지 유도 안구 질환의 임상 치료를 위한 잠재적인 표적 유전자로서 제시하였다. 결론적으로 본 연구의 결과는 1차 쥐의 각막 상피세포가 PM_2.에 의한 각막 상피세포 병리기전 연구에 유용한 모델일 수 있으며, 산화적 및 염증성 반응이 PM_2.유발 안구 표면 장애 유도에 핵심적인 역할을 함을 알 수 있었다.