• Journal of Practical Agriculture & Fisheries Research
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• v.24 no.1
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• pp.14-22
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• 2022

The purpose of this study is to provide evidence for discovering functional materials through the anti-inflammatory efficacy screening of randomly selected medicinal herbs. We prepared 70% ethanol extracts from 10 herbs and evaluated for the inhibitory effect of NO production on LPS-stimulated mouse macrophage cell line Raw 264.7. As a result, it was confirmed that the Chrysanthemi Zawadskii Herba (CZ) extract had the highest effect of inhibiting NO production induced by LPS. We therefore measured and compared NO inhibitory effects at different concentrations (10, 50, 250 ㎍/mL) of 70% ethanol and water extract of CZ. It was observed that both ethanol and water treatment groups inhibited NO production in a concentration-dependent manner in both ethanol and water treatment groups. In particular, it was confirmed that the CZ 70% ethanol extract (99.97%) had a higher NO inhibitory effect than the water extract (93.32%) in the high concentration (250 ㎍/mL) treatment group. There was no effect of CZ extract on cell viability at all concentrations used in the experiment. Moreover, it was shown that CZ ethanol extract remarkably inhibited the expression of VCAM-1 induced by TNF-𝛼, and it was slightly decreased even by treatment with water extract. This study suggests that Chrysanthemi Zawadskii Herba has potential as a functional substance that regulates vascular inflammation.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.36 no.1
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• pp.21-39
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• 2023

Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Goihwa-san water extract(GHS) in vitro & in vivo. Methods : In vitro, we evaluated the anti-inflammatory effect of GHS by comparing the Raw 264.7 cells with 10, 30, 100, 300㎍/㎖ of GHS for 1 hour before Lipopolysaccharide(LPS) to the single LPS treated group. We examined the relative cell viability by MTT assay and the relative level of LPS, Loxoribine(LOX), Peptidoglycan(PGN), Flagellin(FLA)-induced NO production by using Griess reagent and measured relative iNOS protein level and COX-2 protein level by using western blot and Image analyzing system. We measured the production of TNF-α, IL-1β, and IL-6 by each ELISA kits and then measured the relative levels of IκBα, p-IκBα in whole-cell lysate fraction and NF-κB in nuclear fraction by using western blot and Image analyzing system. In vivo, we induced the paw edema by subcutaneous injection of 100㎕/rat CA and measured the swelling volume of paw by using a plethysmometer and then measured the relative iNOS protein level by using western blot. Results : As a result, in vitro, LPS, PGN-induced NO production was significantly inhibited by pretreatment with GHS. GHS reduced LPS, PGN-induced iNOS expression, PGN-induced COX-2 expression and LPS-induced production of cytokine(TNF-α, IL-1β, IL-6). Expression of IκBα was increased by pretreatment with GHS 100㎍/㎖. And the expression of p-IκBα and NF-κB were decreased by pretreatment with GHS 100㎍/㎖. In vivo, CA-induced inflammation rat model was used for the evaluation of the anti-inflammatory effect of GHS. 0.3 or 1.0g/kg of GHS significantly reduced the increases of paw swelling and iNOS expression in paw tissues. Conclusions : These results show that GHS can decrease inflammatory response via inhibition of the NF-κB pathway in vitro. And in vivo, the anti-inflammatory effect suggest the clinical basis of GHS for the treatment of inflammatory diseases.

• The Korea Journal of Herbology
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• v.32 no.3
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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.

• The Korea Journal of Herbology
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• v.33 no.2
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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.

• Korean Journal of Clinical Laboratory Science
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• v.54 no.3
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• pp.209-216
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• 2022

The effect of Saccharomycopsis fibuligera fermentation on the antioxidant and anti-inflammatory activity of Kerria japonica (K. japonica) extracts was studied. First, the antioxidant activity of the fermented extract was measured using the 2,2-diphenyl1-picrylhydrazyl (DPPH) and 2,2'-azinobis (3-ethylbenzthiazoline 6-sulfonic acid (ABTS) methods. Also, the quantification of polyphenols and flavonoids, which are representative components with antioxidant activity, was performed. The results of the DPPH and ABTS assays showed an increase in the antioxidant activity by 14.39% and 21.74%, respectively, due to fermentation. The polyphenol concentration increased by 10.5%, and the flavonoid concentration increased by 100.0%. In the cell experiment, a cytotoxicity test and a nitric oxide (NO) production inhibitory test were performed using RAW 264.7 cells. Both the control group and the fermentation group showed no cytotoxicity. In the NO production inhibition experiment, the fermentation group showed a 6.85% higher inhibition of NO production compared to the control group. When the inhibitory effects of the extracts on inflammatory cytokine production were assessed, the fermentation group showed 12.4% and 23.5% higher inhibition of interleukin (IL)-1β and IL-6 production, respectively, compared to the control group. In conclusion, due to its potential for inhibiting NO and inflammatory cytokine production, fermented K. japonica extracts could be considered a source of anti-inflammatory compounds.

• BMB Reports
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• v.55 no.6
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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.

• The Korea Journal of Herbology
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• v.24 no.3
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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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• v.47 no.4
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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.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2020.08a
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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.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2020.08a
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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.