감마선을 이용한 육종 차조기의 항염증 효과

심부용,박정현,김성규,지중구,Sim, Boo-Yong,Park, Jung-Hyun,Kim, Sung-Kyu,Ji, Joong-Gu 한국응용과학기술학회 2019 한국응용과학기술학회지 Vol.36 No.2

본 연구는 감마선을 이용한 육종 차조기 추출물을 통해 항염증 효능을 평가하고자 하였다. RAW 264.7 세포에서 MTT를 통해 세포 생존율을 평가하였으며, LPS로 유도한 RAW 264.7 세포에서 ROS, NO, 염증성 사이토카인, $NF-{\kappa}B$, COX-2 등을 ELISA, Luminex 및 PCR로 측정하였다. 그 결과, 육종 차조기 추출물은 $25{\mu}g/m{\ell}$ 이하에서 세포독성이 없었으며, LPS로 유도된 RAW 264.7 세포에서 ROS, NO, 사이토카인($IL-1{\beta}$, IL-6, $TNF-{\alpha}$)의 생성을 억제하였다. 또한, $NF-{\kappa}B$, COX-2의 발현을 감소시켜 육종 차조기 추출물은 뛰어난 항염증 효과를 보였다. 이와 같은 결과는 염증 매개체로 인해 발생하는 질환을 개선하기 위한 새로운 건강식품 및 치료제의 원료로 개발될 수 있다. The purpose of this study was to confirmed anti-inflammatory effect the apple Induced by mutants with ${\gamma}-Ray$ extract. Cell viability was assessed by MTT assay using RAW 264.7 cells. The extracts measured through changes in the levels of reactive oxygen species (ROS), nitric oxide (NO), inflammatory cytokines, NF-kB, and COX-2 on LPS-induced RAW 264.7 cells. All test results were analyzed by ELISA reader, Luminex and RT-PCR. In result, the extracts was not toxic below in 25 ug/ml, and extracts was inhibited the productions nitric oxide, ROS, cytokines (IL-1b, IL-6, TNF-a), NF-kB and COX-2 in LPS-induced RAW 264.7 cells. Also, the expression levels were decreased on mRNA of $NF-{\kappa}B$ and COX-2. In other words, Perilla frutescens var. crispa Induced by mutants with ${\gamma}-Ray$ extracts showed significant anti-inflammatory effect. These results may be developed as a raw material for new health food and therapeutics to ease the related to the above mediators.

방사선 형질전환 차조기와 백출 복합추출물이 퇴행성관절염 관련 매개체에 미치는 영향

심부용(Boo-Yong Sim),주인환(In-Hwan Joo)김성규(Sung-Kyu Kim),지중구(Joong-Gu Ji) 한국응용과학기술학회 (구.한국유화학회) 2021 한국응용과학기술학회지 Vol.38 No.2

본 연구는 방사선 형질전환 차조기와 백출복합물(차조기 복합물)이 퇴행성관절염 관련 매개 체에 미치는 영향을 확인하고자 MIA(monosodium iodoacetate)로 퇴행성관절염을 유도한 랫드로 평가 하였다. 차조기 복합추출물을 2주 동안 25, 50, 100 ㎎/㎏/day의 용량으로 경구 투여하고 랫드의 우측 관절 내 공간에 MIA를 주입한 후 동일 용량을 4주 동안 지속 투여하였다. 이후, 혈청 바이오마커와 무릎 관절 분석의 형태학 및 조직병리학적 분석에 기초한 치료 효과를 평가하였다. 대조군 랫드와 비교하 였을 때 차조기 복합추출물은 혈청 내 염증 및 골 대사 마커(TNF-α, MMP-3, COX-2, PGE 2 , COMP, Aggrecan)의 생성량을 유의하게 감소시켰다. 이와는 반대로 연골 흡수 매개체인 CTX-2 생성을 증가시켰으며, 방사선 형질전환 차조기는 무릎 연골과 활막을 효과적으로 보존하였다. 그 결과, 차조기 복합물은 퇴행성관절염 증상을 개선하였다. 따라서, 차조기 복합물은 퇴행성관절염 관리를 위한 식품 소재로 사용될 수 있다. The present study aimed to evaluate the effects of radiation mutant Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi complex extract(Perilla frutescens var. crispa complex extract) on the mediators related to degenerative arthritis in a monosodium iodoacetate-induced rat model of degenerative arthritis. Perilla frutescens var. crispa complex extract was administered orally at doses of 25, 50 or 100 ㎎/㎏/day for 2 weeks before direct injection of monosodium iodoacetate (3 ㎎/50 μl of 0.9% saline) into the intra-articular space of the rats’ right knees. The rats subsequently received the same doses of oral Perilla frutescens var. crispa complex extract for another 4 weeks. It was evaluated that the treatment effects based on serum bio-markers, and morphological and histopathological analysis of the knee joints. Compared with those in negative control rats, the Perilla frutescens var. crispa complex extract treatments significantly reduced the serum levels of inflammation, bone metabolism markers (i.e., TNF-α, MMP-3, COX-2, PGE 2 , COMP, and Aggrecan). Otherwise, it was significantly increased the production of CTX-2 in cartilage absorption mediators. In addition, the Perilla frutescens var. crispa complex extract treatments effectively preserved the knee cartilage and synovial membrane. As a result, it indicates that the Perilla frutescens var. crispa complex extract improved degenerative arthritis symptoms. Thus, the Perilla frutescens var. crispa complex can be used in food material for the management of degenerative arthritis.

가미옥녀전(加味玉女煎)이 db/db 마우스 당뇨(糖尿)모델에서 지질대사(脂質代謝)와 항당뇨(抗糖尿) 효능(效能)에 미치는 영향(影響)

심부용 ( Boo-yong Sim ),김동희 ( Dong-hee Kim ) 대한본초학회 2016 大韓本草學會誌 Vol.31 No.2

Objectives : Abnormal regulation of glucose and impaired lipid metabolism that result from a defective or deficient insulin are the key etiological factor in type 2 diabetes mellitus. The our study investigated the effects of Gamioknyeo-jeon (GO) on blood glucose and lipid metabolism improved by it in db/db mice (a murine model of type 2 diabetes mellitus). Methods : The animals were divided into 3 groups: Normal groups were not-treated C57BL/6 mice; Control groups were treated orally with DW in db/db mice; GO groups were treated orally with GO (200 ㎎/㎏/day) in db/db mice. After mice were treated with GO for 5 weeks, we measured AST, ALT, creatinine, BUN, body weight, food intake, blood glucose, insulin and lipid levels (total cholesterol, HDL cholesterol, and LDL cholesterol and atherogenic index(AI) and cardiac risk factor(CRF). Results : Serum AST, ALT, creatinine, BUN levels were not changed by GO do not show any toxic effects. GO groups were decreased in body weight, food intake and blood glucose level among compared to Control groups. Also, GO groups were found to have atherogenic Index and cardiac risk factor as well as lipid metabolism improvement (total cholesterol and LDL cholesterol decrease). Finally, GO groups were increased the insulin compared to Normal and control groups. Conclusions : We suggest that GO may have the control effects of diabetes mellitus by improving blood glucose control and lipid metabolism.

련산오계(連山烏鷄)가 monosodium iodoacetate로 골관절염(骨關節炎)을 유도(誘導)한 랫드에 미치는 영향(影響)

심부용 ( Boo-yong Sim ),최학주 ( Hak-joo Choi ),지중구 ( Joong-gu Ji ),김동희 ( Dong-hee Kim ) 대한본초학회 2017 大韓本草學會誌 Vol.32 No.2

Objective : The aim of this study is to investigate anti-arthritis activity using a korea`s natural monument No. 265 designate `Yeonsan-Ogye`. In this study, research by using extracts from different concentration of the Yeonsan- Ogye through an MIA-induced arthritis animal model was being conducted in vivo and scientifically verifying the efficacy of medicinal food. Methods : Yeonsan-Ogye was administered 500 ㎎/㎏/day, 1000 ㎎/㎏/day, 2000 ㎎/㎏/day to SD-Rat, where arthritis was induced by monosodium iodoacetate (MIA) at 60 ㎎/㎖. MMP-9, COMP, CTXⅡ, calcitonin and glycosaminoglycan level in serum were measured by ELISA. The changes of relative hind paw weight bearing ratio by Incapacitance Test Meter and The cartilage of meniscus volume was examined and 3-D high-resolution reconstructions of the cartilage of meniscus were obtained using a Micro-CT system. Also, the histopathological analysis of knee was observed by H&E and safranin-O staining. Results : Production of MMP-9, COMP (all groups) and CTXⅡ (500, 1000 ㎎/㎏/day) level in serum was decreased, respectively, in comparison with control. The other way, production of calcitonin (500, 1000 ㎎/㎏/day) and glycosaminoglycan (all groups) level in serum, Hind paw weight bearing ratio (all groups) was increased, espectively, in comparison with control. The cartilage of patella volume in micro-CT increased significantly. In addition, all groups showed a increase in the cartilage volume and proteoglycan. Conclusion : The results for Yeonsan-Ogye showed significant antiarthritis activity in serum and the cartilage. Therefore, it is thought to be that Yeonsan Ogye can be utilized as a variety of new korea medicie and health foods against arthritis diseases.

방사선 육종 차조기와 백출 복합물이 조골세포와 파골세포의 활성에 미치는 영향

심부용(Boo-Yong Sim)지중구(Joong-Gu Ji) 한국응용과학기술학회 (구.한국유화학회) 2021 한국응용과학기술학회지 Vol.38 No.1

본 연구는 방사선 육종 차조기와 백출 복합물의 조골세포 분화 활성 및 파골세포 형성 억제를 조사하였다. 차조기와 백출 복합물은 MG-63 세포에서 ALP 활성 및 arlizarin red 염색을 확인하였고 조골세포 형성의 영향은 RAW 264.7 세포에서 TRAP 활성과 TRAP 염색을 진행하였다. 세포 독성시험에서 차조기와 백출 복합물은 50 ㎍/㎖ 농도 이하에서 안전한 것으로 확인되었다. ALP 활성 및 골석회화 형성 능력은 대조군보다 활성이 낮았으나;파골세포에서 TRAP 활성을 유의적으로 감소시켰으며;효과적으로 TRAP(+) 다핵세포를 억제하였다. 따라서 차조기와 백출 복합물은 골 흡수 억제 활성을 향상시켜 뼈 관련 질환의 예방 및 치료에 효과적인 것으로 보여진다. The effects of the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture on the activities of osteoblast differentiation and the restraint of osteoclast formation were investigated. the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture in the human osteoblast “MG-63” cell;was examined in relation to alkaline phosphatase (ALP) activity and alizarin red stains. In order to observe the effects of osteoclasts formation;we analyzed RAW 264.7 cell tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains. In cytotoxicity testing;it was confirmed that apple extract is safe at a concentration of 50 ㎍/㎖ or less. The ALP activity and Bone calcification formation ability were the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture had a lower activity than that of control group. However the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture significantly reduced activity of TRAP in the RAW 264.7 osteoclastic cell generation and effectively Inhibited the TRAP(+) multinuclear cells. Thus;our results demonstrate that the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture enhances the inhibitory activity of bone-resorption in RAW 264.7 cells. In conclusion;the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture seem to be effective in the prevention and treatment of bone related disorders.

지골피 추출물의 선천 면역 개선 효과

심부용(Boo-Yong Sim),지중구(Joong-Gu Ji) 한국식품저장유통학회 2020 한국식품저장유통학회지 Vol.27 No.2

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.

아토피 피부염을 유발한 마우스에서 백선피지부자복합방의 항염증 효과

심부용 ( Boo Yong Sim ),김성환 ( Sung Hwan Kim ),김동희 ( Dong Hee Kim ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.3

Objectives : In order to investigate the efficacy of BJBB on atopic dermatitis, various anti-inflammatory factors were studied. Methods : In-vitro, inflammatory mediators, such as MTT and nitric oxide were detected after the addition of LPS with or without BJBB in Raw 264.7 cells. In-vivo, in order to verify the effectiveness of BJBB in atopic dermatitis animal model, its role in inflammation factors and histological changes were observed in NC/Nga mice. Results : BJBB showed cell viability of 100% or higher in all concentration in Raw 264.7 cells. BJBB inhibited LPS-induced productions of inflammatory mediators nitric oxide in RAW 264.7cells. BJBB treated group showed significant decrease in the expression of IL-1b, IL-6 and TNF-a by 40%, 80% and 44% respectively. Also the group showed decrease in the transcription of IL-1b, IL-6 and TNF-a mRNA in spleen by 41%, 93% and 39% respectively. BJBB treated group showed significant decrease in WBC, neutrophil, lympocyte and monocytes immune cell ratio in blood by 54%, 63%, 57% and 86% respectively. BJBB treated group showed decrease in the expression of IgG by 39% respectively. Also, infiltration of adipocytes into skin was suppressed and the thickness of epidermis and dermis were relatively decreased in the BJBB treated group. Conclusion : BJBB has an anti-inflammatory effects in NC/Nga mouse. Thus, these results suggested a beneficial effect of BJBB in treatment with Atopic dermatitis and inflammatory.

백선피지부자복합방(白鮮皮地膚子複合方)의 항아토피피부염 효능에 관한 실험적 연구

심부용 ( Boo Yong Sim ),지중구 ( Joong Gu Ji ),이원융 ( Won Yung Lee ),김수정 ( Su Jung Kim ),김효영 ( Hyo Young Kim ),이지영 ( Ji Young Lee ),김동희 ( Dong Hee Kim ) 대전대학교 한의학연구소 2014 혜화의학회지 Vol.22 No.2

In order to investigate the efficacy of BJBB on atopic dermatitis, various immune related factors were studied. The results and conclusions are as follows. Atopic dermatitis symptoms were improved in BJBB treated group and significant decrease in dermatitis index were observed in 13 weeks. ALT, AST and BUN, Cr levels were all with in the normal ranges in BJBB treated group, indicating no induced toxicity. BJBB treated group showed significant decrease in CD4+, CD11b+/Gr-1+ immune cell ratio in dorsal skin by 33% and 62% respectively. BJBB treated group showed significant decrease in the expression of IL-4, IL-5, IL-13 and histamine by 83%, 62%, 53% and 61% respectively. Also the group showed decrease in the transcription of IL-4, IL-5 and IL-13 mRNA in spleen by 41%, 52% and 50% respectively. BJBB treated group showed decrease in the expression of IgE by 57% respectively. The results above indicated that treatment of BJBB improved atopic dermatitis symptoms by immune modulation activity a clinical evidence. thus, BJBB has a potential use as a composition of medicinal plants for treatment against inflammation related disease.

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.