川芎茶調散加味方(천궁다조산가미방) 주정추출물이 산화적 손상 및 염증에 미치는 영향

박지원 ( Ji Won Bak ),심부용 ( Boo Yong Sim ),이해진 ( Hae Jin Lee ),김동희 ( Dong Hee Kim ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.4

The purpose of this study was verification of the anti-inflammation and anti-oxidant effect of Cheongungdajosan-gamibang extract (CG) in mouse macrophage, RAW 264.7 cells. Methods : We have basically using LPS-stimulated RAW 264.7 cells. The cell toxicity was determined by MTT assay. To evaluate the anti-inflammatory effect of Cheongungdajosan -gamibang, amount of nitric oxide(NO) was measured using the NO detection kit and the IL-1β, IL-6 and TNF-α expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). Also, free radical scavenging assay has tested for DPPH and ABTS radical activity as well as the contents of total polyphenol. Results : In this study, 96.6% or higher cell viability was observed in all tested groups from 1, 10, 100 ㎍/㎖ in RAW 264.7 cells. The RAW 264.7 cells were induced by lipopolysaccharide (LPS) and CG 1, 10, 100 ㎍/㎖. The CG decreased nitric oxide (NO) production activity dose dependently, especially at 100 ㎍/㎖ of 55%. The production of IL-1β, IL-6 and TNF-α were decreased by 51%, 78% and 35% in CG treated 100 ㎍/㎖. CG showed dose-dependent suppression activity of reactive oxygen species (ROS) production, especially at 100 ㎍/㎖ of 37%. DPPH radical scavenging activity and ABTS cation decolorization were activated over 86% and 88% in CG at 1,000 ㎍/㎖ concentration. Conclusions : According to the results, we thought that CG showed anti-inflammatory and antioxidant activities on the RAW 264.7 cells in mouse macrophage. Therefore, this research is expected to provide the fundamental data about the natural material analysis of relating to the anti-inflammation and antioxidant.

당귀수산(當歸鬚散) 주정 추출물이 외상성 창상(外傷性 創傷) 병태모델의 회복능력에 미치는 영향

박지원 ( Ji Won Bak ),심부용 ( Boo Yong Sim ),김동희 ( Dong Hee Kim ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.5

Objectives : The purpose of this study was verification of the restoration ability effect of Danggwisusan extract(DG) in wound induced Rat. Methods : It needed to make a scar(around 2 × 2㎠) on the top of the fascia in the back of the rats and then the rats were divided into 4groups(n=6). Control was not treated at all, where as DG was orally medicated DG, Terra was percutaneously applied Terramycin, and DG + Terra was both orally medicated DG and percutaneously applied Terramycin per day for three weeks. Results : 93% or higher cell viability was observed in all tested groups from 1, 10, 100 ㎍/㎖ in RAW 264.7cells. The DG decreased NO and cytokine production activity dose dependently. The production of IL-1β, IL-6 and TNF-α were decreased by 46%, 46% and 40% in DG treated 100 ㎍/㎖. The size of wound was significantly decreasing in DG, Terra, DG + Terra. WBC was significantly reduced in DG and DG + Terra. Monocyte was significantly reduced in DG, Terra and DG + Terra. Neutrophil was also reduced in DG, DG + Terra but not meaningless. The mRNA expression of MMP-1 was significantly reduced in Terra, MMP-2 was significantly reduced in DG, Terra, DG + Terra, and MMP-9 was significantly reduced in DG + Terra. Conclusions : According to the results, we thought that DG showed anti-inflammatory activities on the RAW 264.7cells in mouse macrophage and in adult rat wound. Moreover, the progress of recovery was found visually, heamatologically, genetically and histopathologically. In conclusion, it could be thought that DG has effect on the treating of wound.

가미사물탕(加味四物湯)의 항염증 효능에 관한 연구

박지원 ( Ji Won Bak ),김선빈 ( Seon Bin Gim ),김은아 ( Eun A Kim ),전지애 ( Ji Ae Jun ),이기무 ( Ki Moo Lee ),김동희 ( Dong Hee Kim ) 대전대학교 한의학연구소 2012 혜화의학회지 Vol.20 No.2

In order to investigate the possibility of Kamisamultang(KMSMT) as therapeutic for the treatment of atopic dermatitis(AD), cytotoxicity, anti-oxidant activity, modulatory and suppression activities of KMSMT were tested. 90% or higher cell viability was observed in all tested groups from 25 to 200 ㎍/㎖using Raw 264.7 cells. KMSMT showed dose-dependent DPPH scavenging activity, with more than 80% scavenging activities at 400 and 800 ㎍/㎖concentrations. KMSMT showed dose-dependent suppression activity of reactive oxygen species(ROS) production, especially at 200 ㎍/㎖of 42.6%. KMSMT decreased nitric oxide(NO) production activity dose dependently, expecially at 200 ㎍/㎖of 30.9%. IL-1β, IL-6, MCP-1, TNF-α production rate were decreased by 45.7%, 15.5%, 8.9%, 16.5% respectively when Raw 264.7 cells were treated with LPS and with KMSMT of 200 ㎍/㎖. However, only IL-1β and MCP-1 showed significant changes. The results above strongly suggest the modulatory and suppressive effect of KMSMT. The results above indicate that KMSMT significantly reduces the effect of oxidative and inflammatory cytokines. The use of KMSMT in atopic dermatitis can be widely suggested.

가미계작지모탕(加味桂芍知母湯) 주정 추출물이 류마티스 관절염 인자에 미치는 영향

심부용 ( Boo Yong Sim ),박지원 ( Ji Won Bak ),김동희 ( Dong Hee Kim ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.5

Objectives : The purpose of this study is to prove the effect of Gamikyejakjimo-tang (jiaweiguishaozhim-tang, GK) on rheumatoid arthritis. Methods : We checked viability and measured production of IL-1β, IL-6, IL-17, TNF-α in RAW 264.7 cell after treat by GK. Then we measured rheumatoid arthritis index score of DBA/1 mice with rheumatoid arthritis induced by CIA after GK oral administration, checked IL-1β, IL-6, IL-17, TNF-α and hs-CRP tests in serum. Also we were observed mRNA expression of IL-1β, IL-6, IL-17 and TNF-α in spleen by RT-PCR. Results : GK showed cell viability of 100% or higher in all concentration in RAW 264.7 cells. GK inhibited LPS-induced productions of rheumatoid arthritis mediators cytokine in RAW 264.7cells. GK treated group showed improvement from rheumatoid arthritis at decreased the index score. Also, GK treated group decreased level in serum of IL-1b, IL-6, IL-17, TNF-a and hs-CRP tests by 31%, 35%, 20%, 57% and 58% respectively. Finally, GK treated group showed decrease expression of IL-1β, IL-6, IL-17 and TNF-α mRNA in spleen by 46%, 51%, 25% and 42% respectively. Conclusions : In this study, in-vitro and in-vivo results observed rheumatoid arthritis factors cytokine of IL-1 β, IL-6, IL-17 and TNF-α decrease in RAW 264.7 cells, serum, mRNA expression. Also, GK showed decrease of inflammation figure in hs-CRP tests depending on effect of rheumatoid arthritis. Thus, these results can used as a effective drug of GK for rheumatoid arthritis.

MIA 유도 골관절염 랫드에 Natural Eggshell Membrane (NEM)이 미치는 영향

심부용(Sim, Boo Yong),박지원(Bak, Ji Won),이해진(Lee, Hae Jin),전지애(Jun, Ji Ae),최학주(Choi, Hak Joo),권창주(Kwon, Chang Ju),김화영(Kim, Hwa Young),Kevin J. Ruff,Karsten Brandt,김동희(Kim, Dong Hee) 한국영양학회 2015 Journal of Nutrition and Health Vol.48 No.4

본 연구는 난각막 추출물 (NEM)이 골관절염 동물 모델에서 골관절염 유발 인자에 대한 예방과 개선에 미치는 영향을 조사하고자 실시하였다. 실험동물인 6주령의 랫드에 NEM을 골관절염 유발 2주전부터 농도별 (52 mg/kg, 200 mg/kg, 400 mg/kg)로 경구 투여하였으며, 2주간의 경구 투여 실시 후 정상군 (normal)을 제외한 대조군과 NEM 경구투여군에게 3.0 mg/mL의 농도로 MIA를 주사하여 골관절염을 유발하였다. 골관절염 유발 후 4주간 경구 투여를 지속적으로 실시한 후 혈액 및 관절, 조직 등을 이용하여 골관절염 유발 인자들을 확인하였다. NO 생성량은 NEM200, 400 (mg/kg)에서 유의성 있는 감소를 나타내었으며, PGE2 생성량은 모든 농도에서 유의성 있는 감소를 나타내었다. 사이토카인 IL-1β와 IL-6 생성량은 IL-1β에서는 모든 농도에서 유의성 있는 감소를 보였으나, IL-6에서는 400 mg/kg 농도에서만이 유의성 있는 감소를 나타내었다. hs-CRP 생성량은 모든 농도에서 대조군에 비하여 유의성있는 감소를 나타내었으며, MMPs 생성량과 LTB4 생성량을 모든 농도에서 유의성 있는 감소를 나타내었다. 또한, COMP 및 CTX-II 검사를 통해 골관절염의 진행 억제에 유의성 있는 감소를 나타내었다. 마지막으로 관절과 연골을 micro-CT 및 조직 염색을 실시한 결과, NEM 경구 투여군은 연골량 및 관절 조직의 변형, 연골세포의 손상도가 대조군에 비하여 손상이 적었음을 확인할 수 있었다. 이상의 결과로 미루어 볼 때, NEM은 농도 의존적으로 골관절염유발 인자를 감소시켜 관절 및 연골에 효능이 있음을 제시하고 있다. 이와 같은 결과는 NEM이 골관절염에 대한 예방과 개선 효과를 나타낼 수 있는 건강기능식품의 원료로 활용될 수 있을 것이라 사료된다. Purpose: The aim of this study is to investigate anti-arthritis activity using natural eggshell membrane (NEM). Methods: NEM was administered at 52 mg/kg, 200 mg/kg, and 400 mg/kg to SD-Rat, where arthritis was induced by monosodium iodoacetate (MIA) at 3 mg. NO production in serum was measured using Griess reagent. Cytokines including IL-1β, and IL-6 were measured by Luminex and PGE2, MMP-2, MMP-9, TIMP-1, LTB4, and hs-CRP were measured by ELISA. The cartilage of patella volume was examined and 3-D high-resolution reconstructions of the cartilage of patella were obtained using a Micro-CT system. Results: Production of NO, IL-1β, IL-6, PGE2, MMP-2, MMP-9, TIMP-1, LTB4, and hs-CRP in serum was decreased, respectively, in comparison with control. The cartilage of patella volume increased significantly. In addition, the NEM group showed a decrease in the cartilage of patella, synovial membrane, and transformation of fibrous tissue. Conclusion: The results for NEM showed significant anti-arthritis activity. These results may be developed as a raw material for new health food to ease the symptoms mentioned above.

아토피 피부염 외치치료제 처방 개발을 위한 실험적 연구

김건우 ( Gun Woo Kim ),박지원 ( Ji Won Bak ),심부용 ( Boo Yong Sim ),김동희 ( Dong Hee Kim ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.4

Atotang was composed of 10 kinds of traditional medicinal herb. This research was performed to examine biological effects of Atotang for the development of prescription on atopic dermatitis. Methods : Atotang was extracted with 80% EtOH. Free radical scavenging assay has tested for anti-oxidative activity as well as the contents of total polyphenol. We observed the production of ROS, nitric oxide(NO) and the inflammatory cytokines such as interleukin-1beta(IL-β), IL-6, tumor necrosis factor-alpha(TNF-α), Prostaglandin E2(PGE2) in Raw 264.7 cells stimulated by LPS. We used Disc diffusion method to investigate antibacterial activity on Candida albicans, Staphylococcus aureus and Staphylococcus epidermis. Result : Content of total phenolic compound of Atotang was 36.3 mg/g ext. DPPH and ABTS scavenging activities were 77% and 46% at 200 ug/ml respectively, showing dose-dependent increase. The amounts of ROS and NO in RAW 264.7 cells were decreased by 30% and 19% at 200 ug/ml, respectively, showing dose-dependent decrease. The prodcution of IL-1beta, IL-6 and TNF-alpha in RAW 264.7 cells were decreased dose-dependently by 81%, 67%, and 20% at 200 ug/ml, respectively. Atotang was reduced LPS-stimulated production of PGE2 by 33%. Atotang on C. albicans, S. aureus and S. epidermis was selected by a disc diffusion method and inhibition effect of the Atotang on the growth of S. epidermis was the greatest. Conclusion : The results indicated that Atotang showed biological activities showing anti-oxidant, anti-inflammatory and antibacterial effects. Based on these results, it is concluded that Atotang can be applied to the prescription on atopic dermatitis.

애기땅빈대의 열수 추출 및 에탄올 추출물의 항산화 활성 비교

강란이 ( Ran Yi Kang ),최학주 ( Hak Joo Choi ),박지원 ( Ji Won Bak ),심부용 ( Boo Yong Sim ),이해진 ( Hae Jin Lee ),김동희 ( Dong Hee Kim ) 대전대학교 한의학연구소 2014 혜화의학회지 Vol.23 No.1

Objectives : In this study, the antioxidant activities of the 80% ethanol and hot water extracts of Euphorbia supina Rafinesque were investigated. Methods : We measured total phenol contents, flavonoid contents, 2,2-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, 2,2``-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging assay. The production of reactive oxygen species was measured in LPS-induced RAW 264.7 cells using flow cytometry system. Results : The content of phenol in the hot water extract was 65.529 ± 0.462 mg/g and 126.932 ± 1.894 mg/g in the 80% ethanol extract, and that of flavonoid in the hot water extract was 16.063 ± 0.471 mg/g and 29.159 ± 1.963 mg/g in the ethanol extract. The 80% ethanol extract also showed higher DPPH and ABTS radical scavenging activities (90.8±1.0% and 92.5±0.7%) than the hot water extract (81.5±0.5% and 91.5±0.2%). The production of reactive oxygen species(ROS) was reduced dose-dependently by 80% ethanol and hot water extract at concentration of 1, 10 and 100 ㎍/㎖of RAW 264.7 cells. Conclusion : According to these results, the 80% ethanol extract of Euphorbia supina Rafinesque has a good anti-oxidative effects than the hot water extract. Thus, the 80% ethanol extract of Euphorbia supina Rafinesque may serve as useful natural antioxidants.

가미방기황기탕(加味防己黃기湯) 주정 추출물이 류마티스 관절염 동물병태모델에 미치는 영향

심부용 ( Boo Yong Sim ),최학주 ( Hak Joo Choi ),박지원 ( Ji Won Bak ),김동희 ( Dong Hee Kim ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.6

Objectives : The purpose of this study is to prove the effect of GamiBangkeehwangkee-tang (fangjihuangqi-tang, BHT) ethanol extract on the immunity and rheumatoid arthritis related inflammatory cytokines. Methods : We checked viability in RAW 264.7 cell after treat by BHT. Then we measured inflammatory and immunity factors of DBA/1 mice with rheumatoid arthritis induced by collagen after BHT oral administration. Also, we checked micro-CT image, bone volume and bone inflammation ratio in micro-CT and structural parameter test. Results : BHT showed cell viability of 95% or higher in all concentration in RAW 264.7 cells. BHT treated group decreased level in serum of IgM and IgG test by 36% and 25% respectively. And BHT treated group showed significant decrease in WBC, neutrophil, lymphocyte and monocytes immune cell ratio in blood by 47%, 22%, 56% and 85% respectively. Also, BHT treated group decreased level in serum of IL-1β, IL-6, IL-17, TNF-α and hs-CRP tests by 29%, 33%, 32%, 24% and 56% respectively. Finally, BHT treated group showed increase ratio of bone volume that decrease ratio of bone inflammation. Conclusions : In this study, the results were observed rheumatoid arthritis factors cytokine decrease in serum. And BHT showed immunoglobulin and immune cells ratio decrease in serum and blood. Also, BHT depending on effects of inflammatory and immunity in hs-CRP test, micro-CT, structural parameter test. Thus, these results can used as a effective drug of BHT for inflammation and immunity.

加味淸肌散(가미청기산)이 항산화와 염증성 사이토카인에 미치는 영향

이윤정 ( Youn Jeong Lee ),심부용 ( Boo Yong Sim ),이해진 ( Hae Jin Lee ),박지원 ( Ji Won Bak ),김동희 ( Dong Hee Kim ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.4

Gami-Chunggisan extract (GCE) is one of the oriental traditional medicine. We investigated the antioxidant effect and reduction of pro-inflammatory cytokine as a functional ingredient for cosmetic products from the GCE. Methods : GCE was prepared by extracting with 80% ethanol. We analyzed total polyphenol and antioxidant activities. To evaluate antioxidant activity, we measured 2,2``-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging assay. Also we measured the production of reactive oxygen species (ROS) and nitric oxide (NO) on Raw264.7 cells. We researched reduction of anti-inflammatory cytokines from concentration of GCE on Raw264.7 cells. Results : Total polyphenol quantity of GCE was included 46.6 ㎎/g. The GCE showed ABTS free radical scavenging ability with more than 89% at 1000 ㎍/㎖. In addition the DPPH free radical scavenging ability from the GCE was activated over 93% at 1000 ㎍/㎖. Production of the ROS was decreased by approximately 26%, upon the GCE treatment at concentration of 100 ㎍/㎖. The GCE at 100 ㎍/㎖ concentration showed inhibitory effect on NO production by 38%. Production of IL-1β and IL-6 were decreased by approximately 56% and 36%, respectively upon GCE treatment at 100 ㎍/㎖. Also, production of TNF-α was decreased by approximately 79% at 100 ㎍/㎖. Moreover, the GCE showed inhibitory effects on the expression of the IL-1β, IL-6 and TNF-α genes in LPS-induced Raw 264.7 cells. Conclusions : From the results above, we conclude that the GCE indicated significant antioxidant effects and induced reduction of pro-inflammatory cytokine.

삼원단(三元丹) 에탄올 추출물이 비만유도인자에 미치는 영향

강경수 ( Kyung-su Kang ),이해진 ( Hae-jin Lee ),심부용 ( Boo-yong Sim ),박지원 ( Ji-won Bak ),최학주 ( Hak-joo Choi ),김동희 ( Dong-hee Kim ) 대한본초학회 2015 大韓本草學會誌 Vol.30 No.4

Objectives: In this study, we investigated the biological activities such as anti-obesity using Samwondan ethanol extract (SWD). SWD is a complex with Salicornia herbacea Linnaeus, Saururus chinensis Baill and Houttuynia cordata Thunberg as the main raw material. Methods: The SWD was extracted 80% ethanol. 3T3-L1 preadipocytes were induced adipogenesis by differentiation media with SWD at 1 μg/mL, 10 μg/mL, and 100 μg/mL. Effect of SWD performed using MTT assay, oil red O staining (observation by microscope), and reverse transcription polymerase chain reaction. Also we measured production of triglyceride (TG), fatty acid, and acetyl-CoA carboxylase (ACC). Results: Non-cytotoxicity was in all test group from range of 1 μg/mL to 100 μg/mL on pre-adipocyte. The droplet and production of lipid were decreased significantly by the SWD. And TG was decreased by approximately 89%, 85% and 82%, upon the SWD treatment at concentration of 1 μg/mL, 10 μg/mL, and 100 μg/mL. Moreover, the SWD showed inhibitory effects on the expression of the C/EBP (CCAAT/enhaner binding protein)β, C/EBPα, and peroxisome proliferator-activated receptor γ genes in adipocytes. The SWD at 100 μg/mL concentration showed inhibitory effect on fatty acid production by 79%. Also ACC production were decreased dose-dependently. Conclusions: From the results above, we concluded that the SWD indicated significantly anti-obesity effects.