• 대한본초학회지
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• 제32권3호
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• pp.45-53
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• 2017

Objective : Nelumbo nucifera, its rhizome and semen have been used as a traditional medicine which was studied on antioxidant, hepatoprotective effect, anti-obesity and the others. However, Nelumbinis Flos have not studied. We investigated protective effects on oxidative DNA damage and anti-inflammatory effects of Nelumbinis Flos. Methods : The antioxidant activity was conducted by 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, 2, 2'-Azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) radical scavenging assay and reducing power assay. Total phenolic content was analyzed. Also, phenolic compounds were detected by HPLC/UV. The inhibitory effect on oxidative DNA damage was determined using ${\Phi}X-174$ RF I plasmid DNA cleavage assay. The anti-inflammatory effect of Nelumbinis Flos was measured by the amount of nitric-oxide (NO) produced and protein levels of iNOS, and COX-2 in LPS induced RAW 264.7 cells. Results : The results of DPPH and ABTS radical scavenging activity at $200{\mu}g/m{\ell}$ of extraction were $97.02{\pm}0.88%$ and $96.42{\pm}0.25%$. Reducing power (fold of L-ascorbic acid as control) was $100.14{\pm}0.31$ at $200{\mu}g/m{\ell}$. Total phenol content was $8.70{\pm}0.02mg/g$. Chlorogenic acid, catechin and epicatechin were found by HPLC. Nelumbinis Flos has inhibitory effect in dose-manner against oxidative DNA damage. In addition, it showed the anti-inflammatory effect by suppression of NO production as well as protein levels of iNOS, and COX-2. Conclusion : This study suggested that Nelumbinis Flos showed potential antioxidant and suppression activities of various factors were related in NO produced. Therefore, Nelumbinis Flos as natural plant resources that may help reduce inflammation and alleviate DNA damage.

• 대한본초학회지
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• 제33권2호
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• pp.9-18
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• 2018

Objectives : Hwangryunhaedok-tang (HRHDT) is one of the well-known prescription herbal drugs of Korean herbal medicine, which has been widely used for the treatment of various bacterial and inflammatory diseases. This study was conducted in order to develop the tablet formulations of HRHDT and compare its efficacy with the other commercial formulations. Methods : Corresponding herbal medicines comprising of HRHDT were extracted with water for 3 hr at $95{\sim}100^{\circ}C$ and then vacuum dried. Subsequently, some pharmaceutical excipients such as microcrystalline cellulose, croscarmellose sodium, magnesium stearate, etc were used to prepare the HRHDT tablets. The contents with characterizing components of HRHDT tablet was compared with the HRHDT decoction. The contents of characterizing components were analyzed with HPLC. Furthermore, we investigated the anti-inflammatory and anti-oxidative abilities of two different commercial HRHDT granules (HJP-1 and HJP-2) and were compared with that of the formulated HRHDT tablets. The anti-oxidant properties of HRHDR were studied using the 1,1-diphenyl-2-picryhydrazyl (DPPH) radical, contents of total flavonoid and polyphenol. In addition, based on this result the anti-inflammatory effects have verified by mechanism from LPS- treated Raw264.7 macrophages. Results : The results demonstrated that HRHDT tablets showed more anti-inflammatory and anti-oxidative effects than HJP-1, HJP-2. Moreover, it showed more superior effects in terms of dose, usability and stability than the granules. Conclusion : Hence, we concluded that in order to improve the quality and efficacy of the Korean herbal medicine, it is necessary to develop appropriate methods and establish standardized techniques for the development of good formulations.

• 대한임상검사과학회지
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• 제54권3호
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• pp.209-216
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• 2022

발효된 황매화의 상업적 사용 가능성을 확인하기 위하여 항산화능 실험과 세포 독성, 항염능 실험을 실시하였다. 항산화능 실험에는 DPPH 실험, ABTS 실험, polyphenol 농도 측정, flavonoid 농도 측정을 실시하였다. polyphenol과 flavonoid 농도 측정에서는 발효군이 대조군에 비해 높은 농도를 나타내어, 발효를 통해 추출물의 유효성분 및 추출 수율을 높인다는 것을 확인하였다. DPPH, ABTS에서는 대조군에 비해 발효군의 항산화능이 높은 것을 확인하였다. 이는 S. fibuligera가 발효 과정에서 생산해 낸 효소에 의해 황매화의 세포벽 연화와 유기산과 에탄올 생성에 의한 추출 수율 증가에 의한 요인으로 생각된다. 항염능 실험에서는 세포 독성과 항염능을 알아보았다. 세포독성의 경우 대조군과 발효군 모두 낮은 세포독성을 보였으며, 염증 전달 물질인 NO 생성 억제능의 경우 발효군이 대조군에 비해 통계적으로 유의한 수준의 항염능 증가를 보였다. 염증성 cytokine인 IL-1β, IL-6의 농도를 측정한 결과 200 ㎍/mL 농도에서 각각 48.1±6.2%, 30.4±2.2%의 억제효과를 나타내었다. 이를 통해 발효 황매화가 염증 억제를 위한 물질로서 사용가능함을 보였다.

• BMB Reports
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• 제55권6호
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• pp.275-280
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• 2022

The treatment of atopic dermatitis (AD) is challenging due to its complex etiology. From epidermal disruption to chronic inflammation, various cells and inflammatory pathways contribute to the progression of AD. As with immunosuppressants, general inhibition of inflammatory pathways can be effective, but this approach is not suitable for long-term treatment due to its side effects. This study aimed to identify a plant extract (PE) with anti-inflammatory effects on multiple cell types involved in AD development and provide relevant mechanistic evidence. Degranulation was measured in RBL-2H3 cells to screen 30 PEs native to South Korea. To investigate the anti-inflammatory effects of Parasenecio auriculatus var. matsumurana Nakai extract (PAE) in AD, production of cytokines and nitric oxide, activation status of FcεRI and TLR4 signaling, cell-cell junction, and cell viability were evaluated using qRT-PCR, western blotting, confocal microscopy, Griess system, and an MTT assay in RBL-2H3, HEK293, RAW264.7, and HaCaT cells. For in vivo experiments, a DNCBinduced AD mouse model was constructed, and hematoxylin and eosin, periodic acid-Schiff, toluidine blue, and F4/80-staining were performed. The chemical constituents of PAE were analyzed by HPLC-MS. By measuring the anti-degranulation effects of 30 PEs in RBL-2H3 cells, we found that Paeonia lactiflora Pall., PA, and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. show an inhibitory activity of more than 50%. Of these, PAE most dramatically and consistently suppressed cytokine expression, including IL-4, IL-9, IL-13, and TNF-α. PAE potently inhibited FcεRI signaling, which mechanistically supports its basophil-stabilizing effects, and PAE downregulated cytokines and NO production in macrophages via perturbation of toll-like receptor signaling. Moreover, PAE suppressed cytokine production in keratinocytes and upregulated the expression of tight junction molecules ZO-1 and occludin. In a DNCB-induced AD mouse model, the topical application of PAE significantly improved atopic index scores, immune cell infiltration, cytokine expression, abnormal activation of signaling molecules in FcεRI and TLR signaling, and damaged skin structure compared with dexamethasone. The anti-inflammatory effect of PAE was mainly due to integerrimine. Our findings suggest that PAE could potently inhibit multi-inflammatory cells involved in AD development, synergistically block the propagation of inflammatory responses, and thus alleviate AD symptoms.

• 대한본초학회지
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• 제24권3호
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• pp.39-50
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• 2009

Objectives : Smilacis glabrae rhizoma (SG) has been traditionally used as a herbal medication of musculoskeletal disorders like arthritis, pain, convulsions, and syphilis in traditional Korean medicine. This study was investigated anti-oxidative and anti-inflammatory effect of fractionated extracts of Smilacis Glabrae Rhizoma in Human Umbilical Vein Endothelial Cell (HUVEC). Methods : SG extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of SG onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymethoxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of ICAM-1 and VCAM-1 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results : Our results indicated that fractionated extracts, especially ethyl acetate (EA) extract, significantly inhibited free radical generation, the TNF-$\alpha$-induced intracellular oxidation. Furthermore, the EA extract protected TNF-$\alpha$-induced adhesion to THP-1, expression of adhesion molecules accompanied by an attenuation of IL-6 and IL-8 formation in HUVEC. Conclusions : These results indicate that EA extract of SG have potential as an agent of atherosclerosis and other chronic inflammatory diseases including diabetes, hypertension, and arthritis.

• Restorative Dentistry and Endodontics
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• 제47권4호
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• pp.38.1-38.15
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• 2022

Objectives: This study investigated the cytotoxicity, radiopacity, pH, and dentinal tubule penetration of a paste of 1.0% calcium-doped zinc oxide nanocrystals (ZnO:1.0Ca) combined with propylene glycol (PRG) or polyethylene glycol and propylene glycol (PEG-PRG). Materials and Methods: The pastes were prepared by mixing calcium hydroxide [Ca(OH)₂] or ZnO:1.0Ca with PRG or a PEG-PRG mixture. The pH was evaluated after 24 and 96 hours of storage in deionized water. Digital radiographs were acquired for radiopacity analysis and bubble counting of each material. The materials were labeled with 0.1% fluorescein and applied to root canals, and images of their dentinal tubule penetration were obtained using confocal laser scanning microscopy. RAW264.7 macrophages were placed in different dilutions of culture media previously exposed to the materials for 24 and 96 hours and tested for cell viability using the MTT assay. Analysis of variance and the Tukey test (α = 0.05) were performed. Results: ZnO:1.0Ca materials showed lower viability at 1:1 and 1:2 dilutions than Ca(OH)₂ materials (p < 0.0001). Ca(OH)₂ had higher pH values than ZnO:1.0Ca at 24 and 96 hours, regardless of the vehicle (p < 0.05). ZnO:1.0Ca pastes showed higher radiopacity than Ca(OH)₂ pastes (p < 0.01). No between-material differences were found in bubble counting (p = 0.0902). The ZnO:1.0Ca pastes had a greater penetration depth than Ca(OH)₂ in the apical third (p < 0.0001). Conclusions: ZnO:1.0Ca medicaments presented higher penetrability, cell viability, and radiopacity than Ca(OH)₂. Higher values of cell viability and pH were present in Ca(OH)₂ than in ZnO:1.0Ca.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 춘계학술대회
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• pp.83-83
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• 2020

염증은 물리화학적 자극이나 세균 감염과 같은 외부 자극에 대응하기 위한 생체조직의 방어 반응의 하 나이며 조직이나 장기의 손상을 회복시키는 기전으로서 매우 중요한 역할을 하지만 염증반응이 과도해 질 경우 궤양성 대장염, 기관지염, 천식, 대장암 등을 유발하는 원인으로 작용하기 때문에 염증반응을 조 절하는 항염증제 개발은 매우 중요시되고 있다. 국내 염증개선 관련 시장은 현재 약 3조원 규모이며 지속 적인 성장세에 있지만 천연소재의 기능성 중심으로 출시된 제품 중 다수의 제품에서 주로 수입 원료를 사용하고 있어 국내 자생 및 재배하는 천연 소재들의 효능에 대한 검증을 통해 수입 원료 의존도를 낮추 기 위한 연구가 요구되고 있다. 따라서 본 연구에서는 순창에서 재배되는 참두릅 (Aralia elata Seem)과 도라지 (Platycodon grandiflorum)를 이용하여 천연 염증개선 소재화를 목적으로 기능성 증진을 위한 추 출조건 선정과정과 대식세포에 대한 세포독성 및 항염증 효능을 확인하여 천연소재를 이용한 염증개선 소재화를 시도하고자 한다. 본 연구는 전북 순창에서 재배되는 참두릅과 도라지의 추출조건을 선정하기 위해 용매, 온도 및 시간별 추출물의 total polyphenol 함량 평가를 통하여 최적 추출조건 선정을 진행하 였으며, 선정된 추출조건에서 추출된 추출물의 항산화 활성을 측정하기 위하여 DPPH & ABTS radical scavenging activity 및 total flavonoids 함량을 확인하여 항산화 효능을 평가하였다. 또한 대식세포인 Raw 264.7을 사용하여 MTT assay, Nitric oxide (NO) 생성 억제 효능을 확인하여 세포독성 및 항염증 활 성을 평가하였다. 실험결과, Total polyphenol 함량 분석을 통해 최적 추출조건이 선정된 두릅 (주정 40%, 25℃, 3 h), 도라지 (주정 60%, 25℃, 1 h)의 추출물을 이용하여 DPPH & ABTS radical scavenging activity 및 total flavonoids 함량을 분석한 결과, 도라지보다 두릅에서 더 높은 항산화 활성을 나타내었다. 대식세포를 활용한 두릅과 도라지의 세포독성을 측정한 결과, 100 ug/mL 이내의 농도에서 독성활성이 나타내지 않음을 확인되었으며, 항염증 활성을 측정한 결과 100 ug/mL에서 두릅 추출물이 도라지 추출 물보다 약 33% 이상 NO 생성억제 활성이 높게 나타내었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 춘계학술대회
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• pp.84-84
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• 2020

염증 (Inflammation)은 병원체, 손상된 세포, 자극물질 등으로 인한 손상에 대해 작용하여 조직이나 장기의 손상을 재생하는 작용으로써 신체 방어 기전들 가운데 매우 중요한 역할을 하는 것으로 알려져 있다. 이러한 염증반응이 과다할 경우 각종 염증성 질환 혹은 암 등을 유발하는 원인으로 발전할 수 있어 항염증제의 개발은 전 세계적으로 중요시되고 있다. 선천적 면역을 담당하는 대식세포는 lipopolysaccharide(LPS), 활성산소 (ROS) 등에 의해 자극되어 염증인자 생성에 관여한다. 따라서 본 연구에서는 항염증 소재 개발을 목표로 하며, 국내 천연소재를 활용한 기능성 항염증 소재로 전북 순창군에서 재배된 초석잠(Stachys Sieboldii MIQ.)과 돼지감자 (Helianthus tuberosus L.)를 이용하여 항산화 활성을 평가하고자 하였으며, 대식세포주를 활용한 세포독성 및 항염증 활성에 대한 효능을 확인하고자 하였다. 본 연구는, 전북 순창 지역에서 재배 된 초석잠과 돼지감자를 사용하여 각 조건의 추출 용매, 온도, 시간별 추출물의 Total polyphenol 함량 평가를 통한 최적 추출조건 선정을 진행하고, 선정된 추출조건의 추출물의 항산화 활성을 측정하기 위해 DPPH & ABTS radical scavenging activity 분석 및 Total flavonoids 함량 분석을 통해 항산화 효능 평가를 진행하였다. 또한 항염증 소재로의 활용을 위해 대식세포인 Raw 264.7을 사용하여 농도별 MTT assay를 진행하여 세포독성 평가를 진행하였고, Nitric oxide (NO) 생성억제 효능을 확인하여 항염증 활성을 평가하였다. 실험 결과, Total polyphenol 함량 분석을 통해 최적 추출조건이 선정된 초석잠 (25℃, 주정 60%, 3 h), 돼지감자 (25℃, 주정 40%, 1 h)은 최적 조건에서 약 58 mg GAE/g 및 158 mg GAE/g의 총 폴리페놀 함량을 보였으며, DPPH & ABTS radical scavenging activity 및 Total flavonoids 함량 분석한 결과, 초석잠이 돼지감자보다 더 높은 항산화 활성을 나타내었다. 대식세포 실험에서의 추출물 처리군의 세포독성 측정 결과, 100 ug/mL 이내의 농도에서 독성활성이 나타나지 않음을 확인하였고, Nitric oxide (NO) 생성 억제활성 측정을 통해 LPS 처리군 대비 접종량 100 ug/mL 기준 각각 초석잠 약 47%, 돼지감자 약 49% 수준의 항염증 활성을 확인하였다.

• 생명과학회지
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• 제33권12호
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• pp.978-986
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• 2023

선천면역체계에 의한 염증은 감염에 의해 매개되는 환경적 위험 요인에 대한 보호 메커니즘이며 암 발병을 포함한 다양한 인간 질병의 발병 원인이기도 하다. 지의류는 다양한 질병을 치료할 수 있는 가능성을 지닌 다양한 생체 활성분자를 가지고 있다는 측면에서 점점 더 주목받고 있다. 이끼류의 2차 대사산물이 지닌 항산화, 항염증 그리고 항암 활성에 대해서 널리 보고되었지만 아직까지 구체적인 메커니즘은 밝혀지지 않았다. 본 연구에서는 남극 지의류 Usnea aurantiaco-atra의 메탄올 추출물에 대한 항염증 및 항암 활성의 분자적 메커니즘을 조사하고자 하였다. 본 연구결과에 의하면 메탄올 추출물은 COX-2, IL-6, iNOS, TNF-α 및 NO 생성과 같은 주요 염증 지표들에 대해 농도 의존적으로 조절함으로써 항염증 활성을 나타냈다. 또한, 추출물이 농도 의존적으로 HCT116 결장암 세포에 대해 세포독성 활성을 가지며, caspase-3 활성화에 의한 세포사멸 유도를 통해 암세포의 증식을 현저히 감소시키는 것을 관찰했다. 이 연구에서 남극 이끼류인 Usnea aurantiaco-atra의 메탄올 추출물이 항염증과 항암 활성을 갖는다는 사실을 처음으로 보였으며 이러한 결과는 염증과 암 사이의 연관성을 뒷받침하는 분자 메커니즘에 대한 새로운 통찰력을 보여준다.

• 대한본초학회지
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• 제39권3호
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• pp.57-67
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• 2024

Objectives : The aim of this study is to evaluate the potential biological activity of Veronica incana extracts (VIE) through in vitro, ex vivo, and in vivo experiments. Methods : In vitro, we conducted analyses on the total polyphenol (TP) and total flavonoid (TF) levels, alongside DPPHand ABTS radical scavenging activities. Ex vivo evaluations on adipose tissue measured glycerol release as a marker of lipolysis. In LPS-induced RAW 264.7 cells, we quantified nitric oxide (NO) production. Following H₂O₂ induction in U2OS cells, we performed mitochondrial assays such as MitoSox and MitoTracker. Moreover, Bodipy assays were conducted in 3T3-L1 cells. In vivo, we performed anti-osteoarthritis effect of VIE against monosodium iodoacetate (MIA)-induced osteoarthritis in rats. Results : The results presented encompass a myriad of models, from cell culture to animal experiments as well as ex vivo studies. VIE demonstrated high TP and TF contents, potent DPPH and ABTS scavenging activities, and regulated glycerol release. Moreover, the inhibition of NO production in LPS-induced inflammation was notably confirmed and the reduction of lipid droplets was distinctly shown. Furthermore, in H₂O₂-induced U2OS cells, MitoSox was effectively reduced while MitoTracker noticeably increased. In vivo assays confirmed a significant increase in hindpaw weight distribution (HWD) decreased by MIA after VIE treatment. Additionally, VIE inhibited serum inflammatory cytokines (TNF-𝛼, IL-6, and IL-1𝛽) and MDA levels in joint tissue. Conclusion : In conclusion, Veronica incana exhibited various pharmacological effects including antioxidant, anti-obesity, and anti-inflammatory properties.