[보고서]COPD/천식 등 만성질환 치료용 글로벌 천연물신약 연구사업

오세량

COPD/천식 등 만성질환 치료용 글로벌 천연물신약 연구사업
Development of natural drug candidates for the global level therapeutics against chronic inflammation related diseases such as COPD/asthma and obesity 원문보기

보고서 정보
주관연구기관	한국생명공학연구원 Korea Research Institute of Bioscience and Biotechnology
연구책임자	오세량
참여연구자	이형규 , 안경섭 , 이현준 , 김종평 , 권두환 , 이수의 , 김문옥 , 류형원 , 김정희 , 권옥경 , 김두영 , 이재원
보고서유형	최종보고서
발행국가	대한민국
언어	한국어
발행년월	2018-11
과제시작연도	2018
주관부처	과학기술정보통신부 Ministry of Science and ICT
등록번호	TRKO201900016919
과제고유번호	1711080350
사업명	한국생명공학연구원연구운영비지원(주요사업비)
DB 구축일자	2019-11-09
키워드	천식/만성폐쇄성폐질환.천연물.활성소재.작용기전.대사질환.asthma/COPD.natural product.active material.MoA.metabolic disease.
DOI	https://doi.org/10.23000/TRKO201900016919

초록 ▼

◦ YPL-001의 미국임상 2a에서 유효 단일물질들의 COPD의 임상마커 제시와 MoA 규명으로 임상 2b/3상에서 핵심적인 마커로 활용 가능하여 기업의 아웃소싱의 백업데이터 마련으로 기업의 기술마케팅에서 주도적 역할이 가능해짐.
◦ 천식/COPD 천연물신약 후보소재의 활성물질이 실제 천식환자의 주요 유발인자인 집먼지진드기에 의한 천식 동물모델에서 스테로이드와 동등한 치료효과를 보임으로 경증환자에게 스테로이드 대체요법으로 사용가능함.
◦ Serum amyloid A 는 대표적 천식치료제인 steroid에 의해 유발되는 염증성 물질로 picroside II에 의한 serum amyloid A 억제효과를 규명함으로써 steroid 저항성 천식 또는 steroid 병용치료제로서의 picroside II의 활용 가능성을 시사함.
◦ 천연물의 다양한 면역기전 규명으로 신규 또는 기존의약품 병용/대체용 천연물신약으로 개발함.
◦ P.aeruginosa의 특정 단백질 성질을 이용한 질병관련 타겟 실험 시스템 구축 가능함.
◦ 만성염증/대사질환에 우수 원천소재를 세포 및 동물활성 평가를 통해 발굴하여 관련기업과 블라인드 미팅을 통해 예방/치료용 건강 기능성식품 및 천연물 소재로 기술이전 가능함.

(출처 : 요약서 3p)

Abstract ▼

Ⅳ. Research Results
◦ Identification of the MoA of COPD drug candidate (YPL-001) and active constituents of it
◦ Studies of SAA and IL-33 expression and therapeutic immune mechanism for YPL-001
◦ Development of new botanical raw materials effective in vitro/in vivo for the new drug candidates against asthma/COPD
◦ Development of new botanical raw materials effective in metabolic disease target proteins in vitro/in vivo

(출처 : SUMMARY 5p)

목차 Contents

표지 ... 1
제 출 문 ... 2
보고서 요약서(보고서 초록) ... 3
요 약 문 ... 4
S U M M A R Y ... 5
목차 ... 6
제 1 장 연구개발과제의 개요 ... 7
제 1 절 연구개발의 목적 ... 7
제 2 절 연구개발의 필요성 및 범위 ... 7
제 2 장 국내외 기술개발 현황 ... 16
제 1 절 COPD/천식 치료제 국내외 연구개발 현황 ... 16
제 2 절 만성염증질환 치료제 국내외 연구개발 현황 ... 20
제 3 절 만성호흡기질환 유발 인자 및 조절물질개발 현황 ... 21
제 4 절 대사성질환 치료제 국내외 연구개발 현황 ... 22
제 3 장 연구개발수행 내용 및 결과 ... 25
제 1 절 YPL-001의 COPD 치료제 기전규명 ... 25
제 2 절 만성항염증기반 천식 천연물소재 발굴 ... 54
제 3 절 만성호흡기질환 유바인자 및 조절물질에 관한 연구 ... 215
제 4 절 대사질환 표적단백질 조절 천연물소재 발굴 ... 223
제 4 장 목표달성도 및 관련분야에의 기여도 ... 257
제 1 절 목표달성도 ... 257
제 2 절 관련분야 기여도 ... 258
제 5 장 연구개발결과의 활용계획 ... 259
제 6 장 참고문헌 ... 261
끝페이지 ... 263

표/그림 (300)

표 핵심 지역별 천연물 의약품 정의
표 천연물의약품 산업의 고부가가치성
표 합성의약과 천연물신약 개발 프로세스 비교
표 미국 천연물의약품을 개발 중인 회사들
표 미국의 천연물 의약품 파이프라인
표 중국 TCM 처방 의약품 시장 현황
표 2012년 상위 TA별 유럽 천연물 의약품 시장(USD)
표 합성의약품보다 약효가 뛰어난 천연물의약품
표 천연물 의약품에 대한 Totality of the Evidence 접근
표 세계의 천식 및 만성폐쇄성폐질환(COPD) 시장(Koncept Analytics)
표 MUC5AC의 발현을 유도하는 신호전달 기전
표 인간 기도 상피세포에서 다양한 자극원에 의한 MUC5AC의 발현 유도
표 다양한 자극원에 의해 유도된 MUC5AC promoter에서 YPL-001와 이로부터 분리한 단일화합물 Verproside와 Picroside II에 의한 억제 효과
표 천연물 신약 YPL-001에 의한 PMA에 유도된 MUC5AC 분비량의 저해 효과
표 Verproside에 의한 PMA에 활성화된 PKC isozymes의 억제효과
표 YPL-001(1)과 이로부터 분리된 단일 화합물 (2-8)에 의해 PMA에 활성화된 novel PKC (δ, θ and μ) 의 저해효과
표 Elastase에 의해 유도된 COPD유발 동물모델에서PKCδ 의 발현 증가
표 TNF-α에 의해 유도된 MUC5AC mRNA와 단백질 분비량의 억제 효과. Effect of Iridoid compounds on mRNA and protein expression levels of MUC5AC gene (A) Cell viability of H292 at indicated concentrations of Iridoid compounds for 24 hr. (B) Inhibitory effect of TNF- α-induced MUC5AC gene by each iridoid compounds using qRT-PCR. (B) Inhibitory effect of ELISA of protein expression of MUC5AC in the presence Iridoid compounds (1-11) at 10 μM
표 (A) Chemical structure of piscroside C. (B) Cell viability of NCI-H292 responding to piscroside C was normal. (C) Evaluation of piscroside C on mRNA expression of MUC5AC using RT-qPCR. NCI-H292 cells were pre-treated with each indicated concentration of piscroside C for 2 h and then treated with TNF-α (20 ng/mL) for 12 h. (D) The effect of piscroside C on the secretion of MUC5AC protein was analyzed by ELISA. NCI-H292 cells were pretreated with piscroside C for 2 h before treatment with TNF-α (20 ng/mL) for 24 h. Data means (± S.D.) of at least (*p< 0.05, **p< 0.01, and ***p< 0.001 compared with the control, TNF-α, alone)
표 qRT-PCR을 위한 프라이머 염기서열
표 Inhibitory effects of piscroside C on TNF-α-activated NF-κB activity. (A) The inhibitory effect of piscroside C on MUC5ACtranscription was assayed using a luciferase reporter gene driven by the MUC5AC promoter (NCI-H292/MUC5AC promoter-Luc) in NCI-H292 cells. For each concentration of piscroside C, cells were pre-treated for 2 h, then TNF-α(20 ng/mL) was added and incubated continued for 16 h (*p< 0.05, **p< 0.01, and ***p< 0.001). (B) Reduced binding of NF-κB to the MUC5AC promoter by piscroside C was determined by ChIP using anti-NF-κBp65 antibody. Primers for PCR were designed to identify the NF-κB binding sites within the MUC5ACp romoter region (-464 to -365). NCI-H292 cells were pre-treated with each indicated concentration of piscroside C for 2 h and then treated with TNF-α(20 ng/mL) for 30 min. Rabbit IgG served as a negative control, and input samples (unprecipitated chromatin)served as positive controls for PCR amplification.(C) The effect of piscroside C combined with IKK (BAY11-7082) or TAK1 (5z-7-oxozeaenol) inhibitor on MUC5AC mRNA expression. RT-qPCR was performed in at least. Data means ± SD (; *p< 0.05, **p< 0.01, and ***p< 0.001 compared with the control, TNF-α, alone)
표 Inhibitory effect of piscroside C on TNF-RSC formation. (A) NCI-H292 cells were pre-treated with different concentrations of piscroside C for 2 h, exposed to with TNF-α(20 ng/mL) for 30 min, and then lysed for assays. The anti-TNFR1 antibody immunoprecipitates were immunoblotted (IB) with anti-TRADD, RIP1, TRAF2, TAK1 or TNFR1 antibodies;4% of the whole cell extract for IP was used for input.Protein bands were detected by a LAS-4000 imager. (B) The quantification of immunoreactive band was determined bydFujisoftware normalized to TNFR1 intensity. Data means (± SD, ) of at least (*p< 0.05, **p< 0.01, and ***p< 0.001 compared with the control, TNF-α, alone). (C)Piscroside C had no inhibitory effects on the binding between TNF-α and its receptor, TNFR1. Extracellular interaction between TNF-α ligand and its receptor, TNFR1 (deep blue line) was not affected by the addition of piscroside C (yellow and green line). H292 cells were incubated with biotin-labeled human TNF-α, followed by fluorescein isothiocyanate-labeled streptavidin. The cells were subjected to FACS analysis using FACSCanto II (BD Biosciences, San Jose, CA). The data were analyzed using FlowJo software (TreeStar, Ashland, OR). Negative control preparations were incubated with biotin-labeled soybean trypsin inhibitor (SBTI, red line) or anti-TNF-α antibody-bound TNF-α (light blue line), which does not bind to cell-surface proteins or TNFR1
표 Piscroside C inhibits kinase activity of PKC for IKKα/β activation and interaction between PKCδ and TRAF2. (A) NCI-H292 cells were pretreated at each indicated concentration of piscroside C for 2 h, and then treated with TNF-α(50 ng/mL) for 30 min. Whole cell lysates were assessed by western blotting using antibodies against p-PKC (pan, detecting autophosphorylation at a catalytic domain of PKC), PKC, p-ERK, ERK and Tubulin. Tubulin was the protein loading control. The numbers below the bands indicate the relative band intensity (-fold of control). (B) NCI-H292 cells were pre-treated with the indicated concentration of piscroside C and staurosporine (a PKC inhibitor) for 2 h before treatment of TNF-α(50 ng/mL) for 30 min. Antibodies against p-IKKα/β, IKKβ, p-ERK, and ERK were used for western blotting. The numbers below the bands define the relative band intensity (-fold of control). (C) HEK293T cells were transiently transfected with expression vectors for PKCδ -HA and TRAF2-Flag. Twelve hours after transfection, the cells were pre-treated for 2 h with piscroside C or staurosporine, then subsequently treated with TNF-α(50 ng/mL) for 30 min, then lysed for IP assay using anti-FLAG antibody. Anti-Flag and anti-HA antibodies were used for immunoblotting. The relative band intensities of HA-tagged PKCδ proteins were quantified using Fujisoftware. (D) Predicted piscroside C binding to an ATP binding site of PKCδ using molecular docking simulation. The developed homology model structure was drawn as a white cartoon image. The ATP binding pocket of PKCδis shaded a pink color. Piscroside C (colored green) is drawn as a stick model
표 Schematic depicting the molecular mechanism of piscroside C suppressing TNF-α -induced MUC5AC expression in NCI-H292 cells. Piscroside C has an inhibitory effect on TNF-α/TNF-RSC/IKK/NF-κB pathway by inhibiting PKC activity. Our data suggested that piscroside C has no inhibitory effect on the extracellular interaction between TNF-α ligand and its receptor, TNF-R1 (Ø). TRAF2 activation by PKC is essential for TNF-RSC/IKK/NF-κB activation. We propose that piscroside C inhibits PKC activity, including PKCδ, and its subsequent recruitment to TRAF2, thereby inhibiting IKK/NF-κB activation in airway epithelial cells. We cannot rule out the possibility that piscroside C negatively regulates interactions between TNFR1 and PKC (grey line)
표 COPD 천연물신약 치료제 YPL-001과 picroside II의 LPS-유도 SAA1 발현 억제 효과. (A) Structure of picroside II (PIC II). (B) THP-1 cells were pretreated with YPL-001 (5, 10, 20 ㎍/ ㎖) or PIC II (5, 10, 20 μM) or for 1h and stimulated with LPS (0.2 ㎍/㎖), and then 10 expression levels of SAA1 mRNA were measured by real-time RT-PCR. (C-D) THP-1 cells were pretreated with indicated concentrations of PIC II or DEX (1 μM) for 1h and then stimulated with LPS (0.2 ㎍/㎖) for 6h. The mRNA expressions of SAA1 (C) and TNF (D) were measured by real-time RT-PCR. (E) THP-1 cells were pretreated with indicated concentrations of PIC II for 1h and then stimulated with LPS (0.2 ㎍/㎖) plus DEX (1 μM) for 6h. The mRNA expression of SAA1 was measured by real-time RT-PCR. ##p<0.01 and ###p<0.001, compared with medium alone;*p<0.05, **p<0.01, and ***p<0.001, compared with LPS alone
표 COPD 천연물신약 치료제의 picroside II의 SAA1 의한 IL-33 생성 억제효과. (A) Effect of PIC II on the SAA-induced IL33 mRNA expression was measured by qRT-PCR. NCI-H292 cells were pretreated with indicated concentrations of PIC II or DEX (1μM) for 1h and subsequently treated with SAA (0.5 ㎍/㎖) for 6h. (B) Effect of PIC II on the SAA-induced IL-33 secretion was assayed using ELISA. NCI-H292 cells were pretreated with PIC II or DEX (1 μM) for 1h and subsequently treated with SAA (0.2 ㎍/㎖) for 6h
표 COPD 천연물신약 치료제 picroside II의 SAA1 의한 IL-33 생성 억제 신호전달기전. NCI-H292 cells were pretreated with indicated concentrations of PIC II or DEX (1 μM) for 1h and subsequently treated with SAA (0.5 ㎍/㎖) for 15min. (A) MAPKs (p38 and ERK1/2) and (B) NF-κB (IκBα and p65) phosphorylation were determine by western blot analyses. The expression of unphosphorylated forms of MAPKs and NF-κB is shown as the lading control. The relative protein level ratios of phospho-p38/p38, phospho-ERK1/2/ERK1/2, phospho-IκBα/Iκ Bα, and phosphor-p65/p65 were quantified using Image J software. #p<0.05, ##p<0.01 and ###p<0.001, compared with medium alone; *p<0.01, **p<0.01 and ***p<0.001, compared with SAA alone
표 집먼지 진드기에 의한 천식유발 면역기전
표 (A) A timeline of allergen sensitization, exposure, and drug treatment in this study (DEX; dexamethasone, i.n; intranasal). The total cells (B) and differential cells (C) in BALF of mice were collected at 48h after the last HDM challenge, and quantified in DiffQuick-stained reagent. NC; normal control mice treated with saline only, model; HDM-sensitized/challenged mice, YPL 15 and 30; YPL-001 (15 and 30 ㎎/㎏) + HDM-sensitized/challenged mice, PIC 15 and 30; picroside II (15 and 30 ㎎/㎏)+HDM-sensitized/challenged mice, DEX; dexamethasone + HDM-sensitized/challenged mice. All data are representative of three independent experiments and represented as the mean ± SEM (n=6 mice/group). #p<0.05,##p<0.01,and###p<0.001, compared with normal control (NC);*p<0.05, **p<0.01, and ***p<0.001, compared with model group
표 After the collection of BALF, lung tissue was fixed, sectioned at 4μm and stained with hematoxylin and eosin (H normal control mice treated with saline only, model; HDM-sensitized/challenged mice, PIC 15 and 30; picroside II (15 and 30 ㎎/㎏) + HDM-sensitized/challenged mice, DEX; dexamethasone + HDM-sensitized/challenged mice
표 Serum samples were collected 48h after the last HDM challenge. The levels of (A) total IgE, (B) HDM-specific IgE, and (C) HDM-specific IgG1 were measured using ELISA. NC; normal control mice treated with saline only, model; HDM-sensitized/challenged mice, PIC 15 and 30; picroside II (15 and 30 ㎎/㎏) + HDM-sensitized/challenged mice, DEX; dexamethasone + HDM-sensitized/challenged mice. All data are representative of three independent experiments and represented as the mean ± SEM (n=6 mice/group). ###p<0.001, compared with normal control (NC);*p<0.05, **p<0.01, and ***p<0.001, compared with model group
표 BALF samples were collected from mice 48h after the last HDM challenge. The levels of (A) Th2-related s, (B) Th1-related cytokine, and (C) asthma-related cytokine were measured using ELISA. NC; normal control mice treated with saline only, model; HDM-sensitized/challenged mice, PIC 15 and 30; picroside II (15 and 30 ㎎/㎏) + HDM-sensitized/challenged mice, DEX; dexamethasone + HDM-sensitized/challenged mice. All data are representative of three independent experiments and represented as the mean ± SEM (n=6 mice/group). ###p<0.001, compared with normal control (NC);*p<0.05, **p<0.01, and ***p<0.001, compared with model group
표 The mRNA levels of (A) Th2-related cytokines, (B) Th1-related cytokine, (C) Th17-related cytokine, and (D) asthma-related mediators, were determined by real-time RT-PCR. The data were normalized toGapdh gene expression. NC; normal control mice treated with saline only, model; HDM-sensitized/challenged mice, PIC II; picroside II (30㎎/㎏) + HDM-sensitized/challenged mice, DEX; dexamethasone + HDM-sensitized/challenged mice. All data are representative of three independent experiments and represented as the mean ± SEM (n=6mice/group). ##p<0.01and###p<0.001, compared with normal control (NC);*p<0.05, **p<0.01, and ***p<0.001, compared with model group
표 (A) T-bet and GATA3 mRNA expression were determined by real-time RT-PCR. The data were normalized toGapdh gene expression. (B) T-bet and GATA3 protein were analyzed by western blot. (C) The western blot was quantitated by ImageJ. The levels of T-bet and GATA3 were calculated over GAPDH. NC; normal control mice treated with saline only, model; HDM-sensitized/challenged mice, PIC II; picroside II (30㎎/㎏) + HDM-sensitized/challenged mice, DEX; dexamethasone + HDM-sensitized/challenged mice. All data are representative of three independent experiments and represented as the mean ± SEM (n=6mice/group). #p<0.05,compared with normal control (NC);*p<0.05 and ***p<0.001, compared with model group
표 (A) Cytotoxicity of picroside II was assessed by CCK-8 assay. (B) The cytokine levels of IL-5 and IL-13 were measured using ELISA. (C) Th2-related cytokines (Il5, and Il13) and (D) gata3were determined from the activated Th2 cells by real-time RT-PCR. (E) Western blotting of GATA3 and (F) STAT6 phosphorylation were analyzed from the activated Th2 cells. Each group was quantitated by ImageJ, the levels of GATA3 and p-STAT6 were calculated over GAPDH and STAT6, respectively. Data are presented as mean ± SEM of each group. *p<0.05, **p<0.01, and ***p<0.001 indicate statistically significant difference compared with the control (Th2 cells, alone)
표 COPD에서 IL-33 및 Serum amyloid A (SAA)의 역할
표 COPD 천연물신약 치료제 YPL-001과 picroside II의 LPS-유도 SAA1 발현 억제 효과. (A) Structure of picroside II (PIC II). (B) THP-1 cells were pretreated with YPL-001 (5, 10, 20 ㎍/ ㎖) or PIC II (5, 10, 20 μM) or for 1h and stimulated with LPS (0.2 ㎍/㎖), and then 10 expression levels of SAA1 mRNA were measured by real-time RT-PCR. (C-D) THP-1 cells were pretreated with indicated concentrations of PIC II or DEX (1 μM) for 1h and then stimulated with LPS (0.2 ㎍/㎖) for 6h. The mRNA expressions of SAA1 (C) and TNF (D) were measured by real-time RT-PCR. (E) THP-1 cells were pretreated with indicated concentrations of PIC II for 1h and then stimulated with LPS (0.2 ㎍/㎖) plus DEX (1 μM) for 6h. The mRNA expression of SAA1 was measured by real-time RT-PCR. ##p<0.01 and ###p<0.001, compared with medium alone;*p<0.05, **p<0.01, and ***p<0.001, compared with LPS alone
표 COPD 천연물신약 치료제의 picroside II의 SAA1 의한 IL-33 생성 억제효과. (A) Effect of PIC II on the SAA-induced IL33 mRNA expression was measured by qRT-PCR. NCI-H292 cells were pretreated with indicated concentrations of PIC II or DEX (1μM) for 1h and subsequently treated with SAA (0.5 ㎍/㎖) for 6h. (B) Effect of PIC II on the SAA-induced IL-33 secretion was assayed using ELISA. NCI-H292 cells were pretreated with PIC II or DEX (1 μM) for 1h and subsequently treated with SAA (0.2 ㎍/㎖) for 6h
표 COPD 천연물신약 치료제 picroside II의 SAA1 의한 IL-33 생성 억제 신호전달기전. NCI-H292 cells were pretreated with indicated concentrations of PIC II or DEX (1 μM) for 1h and subsequently treated with SAA (0.5 ㎍/㎖) for 15min. (A) MAPKs (p38 and ERK1/2) and (B) NF-κB (IκBα and p65) phosphorylation were determine by western blot analyses. The expression of unphosphorylated forms of MAPKs and NF-κB is shown as the lading control. The relative protein level ratios of phospho-p38/p38, phospho-ERK1/2/ERK1/2, phospho-IκBα/Iκ Bα, and phosphor-p65/p65 were quantified using Image J software. #p<0.05, ##p<0.01 and ###p<0.001, compared with medium alone; *p<0.01, **p<0.01 and ***p<0.001, compared with SAA alone
표 항염증 천연물 소재의 스크리닝 방법
표 유도활성분획법을 이용한 천연물 원천소재 발굴모식도
표 신이 추출물의 UPLC-Q-TOF확인 조건
표 신이 추출물 및 분획물들의 UPLC-QTof-MS 크로마토그램
표 신이 CHCl3층의 칼럼분획물들(Fr.1-6) UPLC-QTof-MS 크로마토그램
표 신이로부터 분리 및 분석된 유효화합물들 8종의 구조
표 신이로부터 분리된 화합물들 8종 MS 크로마토그램
표 신이로부터 화합물들 7종 분리 모식도
표 담배연기농축액 (CSC; cigarette smoke condensate)에 의해 유도된 IL-6 발현 억제
표 담배연기농축액 (CSC; cigarette smoke condensate)에 의해 유도된 MUC5AC 발현 억제
표 담배연기농축액 (CSC; cigarette smoke condensate)에 의해 유도된 AKT와 ERK의 활성 억제
표 신이 단일화합물 7종은 담배노출로 유발된 만성폐쇄성폐질환 마우스 모델에서 기관지 폐포세척액내 염증세포의 침윤, reactive oxygen species (ROS)와 전염증성 cytokine (TNF-α와 IL-6)의 생성억제 효과
표 신이에서 분리된 단일화합물 7종의 담배연기로 유발된 COPD 마우스 폐조직내 염증세포의 침윤억제
표 Molecular docking results (energy: kcal/mol and docking positionsa)
표 작약 종자추출물의 UPLC-Q-TOF확인 조건
표 작약종자 추출물 및 용매분획물의 UPLC-Q-TOF 크로마토그램
표 작약 종자 추출물의 HSCCC 크로마토그램
표 Partition coefficients (K) and separation factors (a) of target compounds 1-5 in different solvent systems
표 작약 종자 추출물에서 분리된 화합물들 5종의 순도 확인 및 분석결과
표 작약 종자로부터 분리된 PL1의 분광학적 자료
표 작약 종자로부터 분리된 PL2의 분광학적 자료
표 작약 종자로부터 분리된 PL3의 분광학적 자료
표 작약 종자로부터 분리된 PL4의 분광학적 자료
표 작약 종자로부터 분리된 PL5의 분광학적 자료
표 작약 종자에서 분리한 화합물들 5종에 대한 세포독성 결과
표 작약 종자에서 분리한 화합물 4종 NO저해 활성 및 suffruticosol A (2) 항염증활성
표 천식유발 동물모델에서 suffruticosol A의 염증세포 유입 억제효과
표 COPD유발 동물모델에서 suffruticosol A의 cytokines 억제 활성
표 백서향 줄기 추출물의 UPLC-PDA-QTOF-MS 조건
표 백서향 줄기 추출물의 UPLC-PDA-QTOF-MS 크로마토그램
$백서향 줄기 추출물 및 분획물들의 bioassay-guided fractionation 활성결과$ 표 백서향 줄기 추출물 및 분획물들의 bioassay-guided fractionation 활성결과
표 백서향 줄기 CHCl3+EA의 분획물 Frs.1-9의 UPLC-QTOF-MS 크로마토그램
표 백서향 줄기에서 분리된 1-6 화합물들의 MS 및 UV chromatogram
표 백서향 줄기에서 분리된 7-12 화합물들의 MS 및 UV chromatogram
표 백서향 줄기에서 분리된 화합물들의 CHCl3 및 EA층 profile 결과
표 백서향 줄기로부터 분리된 DK1의 분광학적 자료
표 백서향 줄기로부터 분리된 DK2의 분광학적 자료
표 백서향 줄기로부터 분리된 DK3의 분광학적 자료
표 백서향 줄기로부터 분리된 DK4의 분광학적 자료
표 백서향 줄기로부터 분리된 DK5의 분광학적 자료
표 백서향 줄기로부터 분리된 DK6의 분광학적 자료
표 백서향 줄기로부터 분리된 DK7의 분광학적 자료
표 백서향 줄기로부터 분리된 DK8의 분광학적 자료
표 백서향 줄기로부터 분리된 DK9의 분광학적 자료
표 백서향 줄기로부터 분리된 DK10의 분광학적 자료
표 백서향 줄기로부터 분리된 DK11의 분광학적 자료
표 백서향 줄기로부터 분리된 DK12의 분광학적 자료
표 백서향 줄기 추출물, 분획물 및 분리된 순수 단일화합물들의 항염효과
표 백서향 줄기 추출물, 분획물 및 분리된 순수 단일화합물들의 항염효과
표 백서향 추출물과 분획물은 ovalbumin으로 유발된 천식마우스 기관지수축, 전염증성 cytokine (IL-5와 IL-13)과 혈청내 면역글로불린 (IgE)의 생성을 감소
표 백서향 추출물과 분획물은 ovalbumin으로 유발된 천식마우스 기관지폐포세척액내 염증세포의 침윤을 감소
표 백서향 추출물과 분획물은 ovalbumin으로 유발된 천식마우스 폐조직내 염증세포의 침윤억제 효과
표 백서향 추출물과 분획물은 ovalbumin으로 유발된 천식마우스 기관지의 점액분비 억제 효과
표 백서향 추출물과 분획물은 담배노출로 유발된 만성폐쇄성폐질환 마우스 모델에서 기관지폐포세척액내 염증세포의 침윤, reactive oxygen species (ROS)와 전염증성 cytokine (TNF-a와 IL-6)의 생성억제 효과
표 백서향 추출물과 분획물은 담배연기로 유발된 COPD 마우스 폐조직내 염증세포의 침윤억제 효과
표 Multivariate statistical analyses for five Korean mistletoes. (A) Dendrogram, (B) PCA score plot, and (C) loading plot. [VC, Viscum coloratum (Komarov) Nakai f. coloratum; VCr, Viscum coloratum (Komarov) Nakai f. rubroaurantiacum (Makino) Kitagawa; KJ, Korthalsella japonica (Thunb.) Engl.; LT, Loranthus tanakae Franch. et Sav.; TY, Taxillus yadoriki (Sieb. ex Maxim.) Danser]
표 (A) Total ion current chromatograms combined with APCI (+) and ESI (-) modes and (B) the charged aerosol detector (CAD) chromatograms of five Korean mistletoe types
표 Cell viability and promoter activity on human PAI-1 gene of crude extracts of five Korean mistletoe. (A) Effect of the crude extract on cell viability. HMC-1 cells were incubated in the presence or absence of 0–40 mg/mL of crude extract, and cell viability was determined using the MTT assay. The data are the mean±SD of three independent experiments. *Significant difference from control (*, p < 0.05, **, p < 0.01, ***, p < 0.001). (B) The HMC-1-pPAI-1-luc cells were pretreated with phytochemicals for 2 h, then stimulated with 0.02 μM of PMA plus 0.5 μM of A23187 in the culture medium for the indicated time. The measurement of luciferase activity was performed 8 h after the stimulation of PMA and A23187. Relative promoter activity (measured by relative light units) is given as x-fold induction of the HMC-1-PAI-1-luc cell without stimulation. #Significant difference from basal expression. *Significant difference from PMA and A23187 treatment only (#, p<0.01, *, p<0.05, **, p<0.01)
표 Cell viability and promoter activity of human PAI-1 gene by oleanolic acid. (A) Effect of oleanolic acid on cell viability. HMC-1 cells were incubated in the presence or absence of 0–80 μM of oleanolic acid, and cell viability was determined using the MTT assay. The data are the mean±SD of three independent experiments. *Significant difference from control (*, p < 0.05, **, p < 0.01, ***, p < 0.001). (B) The HMC-1-pPAI-1-luc cells were pretreated with oleanolic acid (0-20 μM) for 2 h and then stimulated with 0.02 μM of PMA plus 0.5 μM of A23187 in the culture medium for indicated time. Luciferase activity was measured 8 h after the stimulation of PMA and A23187. The data are presented as x-fold induction of the test conditions divided by the activity in basal (untreated) condition. Values are mean±SD of triplicate experiments. #Significant difference from basal expression. *Significant difference from PMA and A23187 treated only (#,p<0.01,*,p<0.05,**,p<0.01)
표 떡쑥 추출물의 UPLC-Q-TOF확인 조건
표 떡쑥 추출물 및 용매분획물들의 UPLC-Q-TOF 크로마토그램
표 떡쑥 추출물의 MPLC 크로마토그램
표 떡쑥 추출물 및 용매분획물의 항염효과
$Identification of phytochemicals by UPLC-PDA-QTof-MS from EtOAc fraction of G. affine$ 표 Identification of phytochemicals by UPLC-PDA-QTof-MS from EtOAc fraction of G. affine
표 떡쑥 용매분획물의 UPLC-QTOF-MS profiling 결과
표 떡쑥으로부터 예상화합물들 8종 MS 및 MS/MS 크로마토그램
표 떡쑥으로부터 생리활성물질 분리 모식도
표 떡쑥로부터 분리된 화합물1의 분광학적 자료
표 떡쑥로부터 분리된 화합물2의 분광학적 자료
표 떡쑥로부터 분리된 화합물3의 분광학적 자료
표 떡쑥로부터 분리된 화합물4의 분광학적 자료
표 떡쑥로부터 분리된 화합물5의 분광학적 자료
표 떡쑥로부터 분리된 화합물6의 분광학적 자료
표 떡쑥로부터 분리된 화합물7의 분광학적 자료
표 떡쑥로부터 분리된 화합물8의 분광학적 자료
표 떡쑥 EA층에서 분리한 단일화합물 8종에 대한 항염효과
표 Effects of luteolin-4′-O-b-D-(6″-E-caffeoyl)-glucoside (7) on iNOS and COX-2 protein expression in LPS- stimulated RAW264.7 cells
$Inhibitory effects of partitioned fractions and compounds (1−8) on HNE activity$ 표 Inhibitory effects of partitioned fractions and compounds (1−8) on HNE activity
표 (A) Effects of compounds 1–8 on the activity of human neutrophils elastase for proteolysis of N-methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide
표 Lineweaver-Burk and Dixon plot for inhibition of compounds 1, 2 and 4-7 on human neutrophils elastase for proteolysis of a substrate
표 개구리밥 추출물의 UPLC-Q-TOF-MS확인 조건
표 개구리밥 추출물의 UPLC-Q-TOF-MS 크로마토그램
표 개구리밥 추출물의 perp-HPLC 크로마토그램
표 개구리밥으로부터 생리활성물질 분리 모식도
표 개구리밥 추출물의 용매분획물들 UPLC-PDA 크로마토그램
표 개구리밥으로부터 분리 및 분석된 유효화합물들 7종의 구조
표 개구리밥으로부터 분리된 화합물들 7종 MS 및 MS/MS 크로마토그램
$Identification of phytochemicals by UPLC-PDA-QTof-MS from BuOH fraction of S. polyrhiza$ 표 Identification of phytochemicals by UPLC-PDA-QTof-MS from BuOH fraction of S. polyrhiza
표 개구리밥 추출물 및 용매분획물의 in vitro 아토피저해 효과
표 개구리밥에서 분리된 단일성분 7종의 in vitro 아토피저해 효과
표 DBA/1 마우스에 개구리밥 추출물 처리 후 몸무게 및 개구리밥 추출물의 소양증 억제 효과
표 DNCB를 처리한 마우스에 대한 개구리밥 추출물의 사이토카인 억제 효과
표 황해쑥 추출물의 UPLC-Q-TOF확인 조건
표 황해쑥 유효분획물의 UPLC-QTof-MS 크로마토그램
표 황해쑥 EA층, CHCl3층을 칼럼분획물들 UPLC-PDA-QTof-MS 크로마토그램
표 황해쑥으로부터 분리 및 분석된 유효화합물들의 구조
표 황해쑥으로부터 분리/분석된 화합물들 13종 MS, MS/MS, UV 크로마토그램
표 황해쑥으로부터 분리/분석된 화합물들 13종 MS, MS/MS, UV 크로마토그램
표 황해쑥으로부터 분리 및 분석된 천연물성분들의 분광학적 데이터
표 황해쑥에서 분리한 화합물 1,3-dicaffeoylquinic acid의 분광학적 자료
표 황해쑥에서 분리한 화합물 1,4-dicaffeoylquinic acid의 분광학적 자료
표 황해쑥에서 분리한 화합물 Luteolin의 분광학적 자료
표 황해쑥에서 분리한 화합물 Jaceosidin의 분광학적 자료
표 황해쑥에서 분리한 화합물 dehydromatricarin의 분광학적 자료
표 황해쑥에서 분리한 화합물 centaureidin의 분광학적 자료
표 황해쑥에서 분리한 화합물 eupatilin의 분광학적 자료
표 황해쑥에서 분리한 화합물 casticin의 분광학적 자료
표 황해쑥에서 분리한 화합물 artanomaloide의 분광학적 자료
표 황해쑥에서 분리한 화합물 8-O-acetylarteminolide의 분광학적 자료
표 Artemisia argyi declined inflammatory cell counts in the BALF. NC: no treatment and no OVA challenge; OVA: OVA challenge; DEX: dexamethasone 3 mg/kg per day and OVA challenge; TOTAL50 and 100: 50 and 100 mg/kg of Artemisia argyi per day, respectively, and OVA challenge; DA10 and 20: dehydromatricarin A 10 and 20 mg/kg per day, respectively and OVA challenge. The values shown as the means ± SD. #P < 0.05 vs NC; *P < 0.05 vs OVA
표 Artemisia argyi reduced airway hyperreponsiveness. NC: no treatment and no OVA challenge; OVA: OVA challenge; DEX: dexamethasone 3 mg/kg per day and OVA challenge; TOTAL50 and 100: 50 and 100 mg/kg of Artemisia argyi per day, respectively, and OVA challenge; DA10 and 20: dehydromatricarin A 10 and 20 mg/kg per day, respectively and OVA challenge. The values shown as the means ± SD. #P < 0.05 vs NC; *P < 0.05 vs OVA
표 Artemisia argyi decreased level of allergen specific Ig E and Th2 cytokines. (A) IL-4, (B) IL-5, (C) IL-13, (D) OVA-specific IgE. NC: no treatment and no OVA challenge; OVA: OVA challenge; DEX: dexamethasone 3 mg/kg per day and OVA challenge; TOTAL50 and 100: 50 and 100 mg/kg of Artemisia argyi per day, respectively, and OVA challenge; DA10 and 20: dehydromatricarin A 10 and 20 mg/kg per day, respectively and OVA challenge. The values shown as the means ± SD. #P < 0.05 vs NC; *P < 0.05 vs OVA
표 Artemisia argyi decreased MMP-9 and phosphorylation of Erk expression on the protein levels. (A) protein level of MMP-9 expression, (b) protein level of phosphorylation of Erk. NC: no treatment and no OVA challenge; OVA: OVA challenge; DEX: dexamethasone 3 mg/kg per day and OVA challenge; TOTAL50 and 100: 50 and 100 mg/kg of Artemisia argyiper day, respectively, and OVA challenge; DA10 and 20: dehydromatricarin A 10 and 20 mg/kg per day, respectively and OVA challenge. The values shown as the means ± SD. #P < 0.05 vs NC; *P < 0.05 vs OVA
표 Artemisia argyi reduced inflammatory cell infiltration and mucus production. Hematoxylin and eosin (H OVA: OVA challenge; EX: dexamethasone 3 mg/kg per day and OVA challenge; TOTAL50 and 100: 50 and 100 mg/kg of Artemisia argyi per day, respectively, and OVA challenge; DA10 and 20: dehydromatricarin A 10 and 20 mg/kg per day, respectively and OVA challenge. The values shown as the means ± SD. #P < 0.05 vs NC; *P < 0.05 vs OVA
표 황칠나무 잎 추출물의 UPLCーCAD와 PDA-QTof-MS확인 조건
표 황칠나무 잎 추출물, CHCl3 분획물, 단일성분의 UPLC-CAD 및 MS 크로마토그램
표 황칠나무 잎 추출물 및 용매분획물의 UPLC-CAD 크로마토그램
표 황칠나무에서 순수 단일성분 분리 모식도
표 황칠나무 잎에서 분리된 DM1의 분광학적 자료
표 황칠나무 잎에서 분리된 DM2의 분광학적 자료
표 황칠나무 잎에서 분리된 DM3의 분광학적 자료
표 황칠나무 잎에서 분리된 DM4의 분광학적 자료
표 황칠나무 잎에서 분리된 DM5의 분광학적 자료
표 황칠나무 잎 추출물, 분획물, 단일성분들의 항염활성 효과
표 Effect of methanolic extract and isolated polyacetylene of Dendropanax morbifera leaves on the infiltration of (A) neutrophils and (B) macrophages in BALF of CS and LPS-induced mice. The cells count was measured using the Diff-Quick staining reagent (magnification, ×400). The values expressed as mean ± S.D. (n = 6). NC, normal control mice with PBS only; CS + LPS, cigarette smoke (CS) and lipopolysaccharide (LPS); ROF, roflumilast (10 mg/kg) + CS + LPS; T 10, (D. morbifera leaves extract, 10 mg/kg) + CS + LPS; C 10, [(9Z, 16S)-16-Hydroxy-9,17-octadecadiene-12,14-diynoic acid, 10 mg/kg] + CS + LPS. Data are expressed as mean ± S.D. #p < 0.01 indicates statistically significantly different from normal control group. *p < 0.05 and **p < 0.01 indicate statistically significant difference compared to CS + LPS alone
표 Effect of methanolic extract and isolated polyacetylene of Dendropanax morbifera leaves on the levels of ROS, NE and proinflammatory cytokines in BALF. (A) ROS production, (B) NE activity, (C) TNF-a and (D) IL-6 release. Data are expressed as mean ± S.D. #p < 0.01 indicates statistically significantly different from normal control group. *p < 0.05 and **p < 0.01 indicate statistically significant difference compared to CS + LPS alone
표 Effect of methanolic extract and isolated polyacetylene of Dendropanax morbifera leaves on the recruitment of inflammatory cells and on mucus secretion in lung tissue. Peribronchial lesion (magnification, ×100) in the lung tissues was stained with H&E solution
표 Effect of methanolic extract and isolated polyacetylene of Dendropanax morbifera leaves on mucus secretion in lung tissue. Mucus secretion was determined by PAS staining (magnification, ×400). Black arrows indicate mucus secretion
표 Effect of methanolic extract and isolated polyacetylene of Dendropanax morbifera leaves on MAPK and NF-кB activation in lung tissue. The levels of ERK, p38, NF-кB and I кB phosphorylation were examined using Western blot analysis. Data are expressed as mean ± S.D. #p < 0.01 indicates statistically significantly different from normal control group. *p < 0.05 and **p < 0.01 indicate statistically significant difference compared to CS + LPS alone
표 Coumestrol 화합물의 구조, UV, MS, HRESIMS 분석결과
표 추출물은행에서 제공받은 콩과 소재의 분석리스트
표 UPLC-QTof-MS를 이용한 콩과소재의 coumestrol 성분 함유확인 결과
표 UPLC-QTof-MS를 이용한 콩과소재의 coumestrol 성분 함유확인 결과
표 콩과작물로부터 분리한 coumestrol의 분광학적 자료
표 Effect of CM on the infiltration of neutrophils and macrophages in the BALF of ALI mice. The BALF differential cell count was performed by Diff-Quikꠙ staining reagent according to the manufacturer’s instructions. NC, normal control mice with PBS; vehicle, 2% DMSO and 2% Tween 20 in PBS; LPS, lipopolysaccharide (10 μg per mouse); DEX, dexamethasone (3 mg/kg) + LPS; CM 10, coumestrol (10 mg/kg) + LPS; CM 20, coumestrol (20 mg/kg) + LPS. Data are expressed as means ± S.D. #p < 0.01 indicates statistically significantly different from vehicle group. *p < 0.05 and **p < 0.01 indicate statistically significant difference compared with the LPS group
표 Effect of CM on the production of ROS, pro-inflammatory cytokines and NO. (A) ROS production was investigated with 20 lM DCF-DA. The levels of (A) TNF-α and (B) IL-6 were measured using the ELISA assay. The absorbance was measured at 450 nm using a microplate reader. (C) NO production was measured in the serum using the Griess reagent. Data are expressed as means ± S.D. #p < 0.01 indicates statistically significantly different from vehicle group. **p < 0.01 indicates a statistically significant difference compared with the LPS group
표 Effect of CM on the recruitment of inflammatory cells and expression of MCP-1 in the lung of ALI mice. (A) Peribronchial lesion (magnification, ×400) in lung tissue was stained with H&E solution. (B) MCP-1 expression was examined using Western blot analysis. Data are expressed as means ± S.D. #p < 0.01 indicates statistically significantly different from vehicle group. **p < 0.01 indicates a statistically significant difference compared with the LPS group
표 Effect of CM on the activation of NF-κB and expression of SOD3 in the lung of ALI mice. The phosphorylation of (A) IкB and NF-кB p65 and expression of (B) SOD3 were detected using Western blot analysis. Data are expressed as means ± S.D. #p < 0.01 indicates statistically significantly different from vehicle group. *p < 0.05 and **p < 0.01 indicate a statistically significant difference compared with the LPS group
표 Effect of CM on the production of pro-inflammatory mediators and activation of NF-κB in LPS-stimulated RAW 264.7 macrophage cells. (A) Effect of CM on the viability of RAW264.7 macrophages. The cells were treated with CM (2, 4 and 8 lM) for 1 h prior to incubation with LPS (500 ng/ml) and were further incubated for 20 h. No noticeable cell death was observed by CM. (B–D) The levels of NO and pro-inflammatory cytokines were determined by the NO assay and ELISA assay. (E) The cells were treated with CM for 1 h prior to incubation with LPS and were further incubated for 30 min. The levels of IкB and NF-κB phosphorylation were detected using Western blot analysis. Quantitative analysis was performed by densitometric analysis. β-actin was used as an internal control. Data are expressed as means ± S.D. #p < 0.01 indicates statistically significantly different from vehicle group. *p < 0.05 and **p < 0.01 indicate a statistically significant difference compared with the LPS group
표 참오동나무 열매 추출물의 UPLC-Q-TOF확인 조건
표 참오동나무 열매 추출물 및 용매분획물의 UPLC-Q-TOF 크로마토그램
표 참오동나무 추출물의 MPLC 크로마토그램
표 참오동나무 열매에서 분리된 PT 1의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 2의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 3의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 4의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 5의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 6의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 7의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 8의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 9의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 10의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 11의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 12의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 13의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 14의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 15의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 16의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 17의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 18의 분광학적 자료
표 참오동나무 열매에서 분리된 PT 19의 분광학적 자료
표 참오동나무 열매에서 분리한 단일성분 20종의 순도 크로마토그램
표 참오동나무 열매에서 분리한 단일성분 20종의 순도 크로마토그램
표 Inhibitory Effects of Isolated Compounds 1−19 on HNE Enzyme Activity
표 Concentration-dependent inhibition and graphical kinetic determination for the isolated compounds. Double-reciprocal Lineweaver−Burk plots for the inhibition of isolated compounds 3, 4, 8−11, 13, 14, 16, 17, and 19 on the HNE-catalyzed hydrolysis of MeOSuc-AAPV-pNA. The plot is expressed as 1/velocity versus 1/substrate (μM−1) without or with an inhibitor in the reaction solution. Each point represents the mean value of three replicates at substrate concentration
표 참오동 열매 추출물에서 분리한 화합물의 항염효과
$Effects of the methanolic extract, solvent fractions, and isolated new compounds of Paulownia tomentosa mature fruit on cell viability, IL-6, and -8 production in TNF-a-stimulated A549 cells. Bar graphs represent means ± S.D. of three independent experiments (*p < 0.05, **p < 0.01, and ***p < 0.001 compared with the control, TNF-a alone)$ 표 Effects of the methanolic extract, solvent fractions, and isolated new compounds of Paulownia tomentosa mature fruit on cell viability, IL-6, and -8 production in TNF-a-stimulated A549 cells. Bar graphs represent means ± S.D. of three independent experiments (*p < 0.05, **p < 0.01, and ***p < 0.001 compared with the control, TNF-a alone)
표 참오동나무 열매 추출물과 분획물은 ovalbumin으로 유발된 천식마우스 기관지 수축, 전염증성 cytokine (IL-5와 IL-13)과 혈청내 면역글로불린 (IgE)의 생성억제 효과
표 참오동나무 열매 물과 분획물은 ovalbumin으로 유발된 천식마우스 기관지폐포세척액내 염증세포의 침윤 억제 효과
표 참오동나무 열매 추출물과 분획물은 ovalbumin으로 유발된 천식마우스 폐조직내 염증세포의 침윤 억제 효과
표 참오동나무 열매 추출물과 분획물은 ovalbumin으로 유발된 천식마우스 기관지의 점액분비 억제효과
표 참오동나무 열매 추출물과 분획물은 담배노출로 유발된 만성폐쇄성폐질환 마우스 모델에서 기관지폐포세척액내 염증세포의 침윤, reactive oxygen species (ROS)와 전염증성 cytokine (TNF-α와 IL-6)의 생성억제 효과
표 참오동나무 열매 추출물과 분획물은 담배연기로 유발된 COPD 마우스 폐조직내 염증세포의 침윤억제 효과
표 참오동나무 줄기 추출물의 UPLC-Q-TOF확인 조건
표 참오동나무 줄기 추출물의 UPLC-Q-TOF 크로마토그램
표 참오동나무 줄기로부터 분리된 verbascoside의 분광학적 자료
표 참오동나무 줄기로부터 분리된 isoverbascoside의 분광학적 자료
표 The extract of P. tomentosa stem bark (PTBE) attenuates the releases of inflammatory cytokines in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The cells were pretreated with PTBE (2.5, 5, 10, 20, 40 and 80 μg/ml) 1 h prior to incubation with LPS (500 ng/ml) for 24 h. (A) No noticeable cell death was observed up to 40 μg/ml of PTBE. (B-C) The concentration of IL-6 and TNF-α in culture media was determined using ELISA. Data are expressed as the mean ± S.D. #p < 0.01 indicates statistically significantly different from normal control group. *p < 0.05 and **p < 0.01 indicates statistically significant difference compared to the LPS group
표 The extract of P. tomentosa stem bark (PTBE) suppresses the recruitment of (A) neutrophils and (B) macrophages in the bronchoalveolar lavage fluid (BALF) of mice with lipopolysaccharide (LPS)-induced ALI. The BALF differential cells count was performed by Diff-Quik® staining reagent according to the manufacturer
표 The extract of P. tomentosa stem bark (PTBE) inhibits the production of ROS and pro-inflammatory cytokines in the BALF of ALI mice. (A) ROS production was investigated with 20 μM of DCF-DA. (B-C) The levels of IL-6 and TNF-α were measured using ELISA assay. The absorbance was measured at 450 nm using microplate reader. Data are expressed as the mean ± S.D. #p< 0.01 indicates statistically significantly different from normal control group. **p < 0.01 indicates statistically significant difference compared to the LPS group
표 The extract of P. tomentosa stem bark (PTBE) reduces the levels of NO and iNOS in the ALI mice (A) NO production was measured in the serum using the Griess reagent. (B) The expression of iNOS was determined using Western blot analysis. Data are expressed as the mean ± S.D. #p < 0.01 indicates statistically significantly different from normal control group. *p < 0.05 and **p < 0.01 indicates statistically significant difference compared to the LPS group
표 The extract of P. tomentosa stem bark (PTBE) inhibits the infiltration of inflammatory cells and the expression of MCP-1 in the lung of ALI mice. (A) Peribronchial lesion (magnification, ×400) in the lung was stained with H&E solution. (B) MCP-1 expression was investigated using Western blot analysis. Data are expressed as the mean ± S.D. #p< 0.01 indicates statistically significantly different from normal control group. *p < 0.05 and **p < 0.01 indicates statistically significant difference compared to the LPS group
표 The extract of P. tomentosa stem bark (PTBE) suppresses the NF-κB activation in the lung of ALI mice. (A) The phosphorylation of IκB and NF-κB p65 were detected using Western blot analysis. (B) Quantitative analysis was performed by densitometric analysis. Data are expressed as the mean ± S.D. #p < 0.01 indicates statistically significantly different from normal control group. *p < 0.05 and **p < 0.01 indicates statistically significant difference compared to the LPS group
표 The extract of P. tomentosa stem bark (PTBE) attenuates the reduced expression of SOD3 in the lung of ALI mice. The expression of SOD3 were measured using Western blot analysis. Data are expressed as the mean ± S.D. #p< 0.01 indicates statistically significantly different from normal control group. *p < 0.05 and **p < 0.01 indicates statistically significant difference compared to the LPS group
표 두메닥나무 추출물의 UPLC-Q-TOF확인 조건
표 두메닥나무 추출물의 UPLC-Q-TOF 크로마토그램
표 두메닥나무 추출물의 MPLC 크로마토그램
표 두메닥나무 추출물 및 칼럼분획물의 UPLC-PDA 및 유효성분의 MS, MS/MS, UV 크로마토그램
표 두메닥나무 추출물 및 칼럼분획물의 UPLC-PDA 및 유효성분의 MS, MS/MS, UV 크로마토그램
표 두메닥나무 추출물에서 분석된 지표 및 유효성분들의 분광학적 데이터
$Effect of a biflavonoid fraction (BF) obtained from extract of D. pseudomezereum on the production of pro-inflammatory mediators in LPS-stimulated RAW 264.7 macrophage cells. The cells were treated with BF (2.5, 5, 10 and 20 μM) for 1 h prior to incubation with LPS (500 ng/ml) and were further incubated for 20 h. (A–C) The levels of pro-inflammatory cytokines and NO were determined by the ELISA assay and NO assay. (D) No noticeable cell death was observed by BF. Data are expressed as means ± S.D. #p < 0.01 indicates statistically significantly different from vehicle group. *p < 0.05 and **p < 0.01 indicate a statistically significant difference compared with the LPS group$ 표 Effect of a biflavonoid fraction (BF) obtained from extract of D. pseudomezereum on the production of pro-inflammatory mediators in LPS-stimulated RAW 264.7 macrophage cells. The cells were treated with BF (2.5, 5, 10 and 20 μM) for 1 h prior to incubation with LPS (500 ng/ml) and were further incubated for 20 h. (A–C) The levels of pro-inflammatory cytokines and NO were determined by the ELISA assay and NO assay. (D) No noticeable cell death was observed by BF. Data are expressed as means ± S.D. #p < 0.01 indicates statistically significantly different from vehicle group. *p < 0.05 and **p < 0.01 indicate a statistically significant difference compared with the LPS group
표 BF suppresses the recruitment of (A) eosinophils (B) macrophages and (C) neutrophils in the bronchoalveolar lavage fluid (BALF) of mice with ovalbumin (OVA)-induced allergic asthma. The BALF differential cells count was performed by Diff-Quik® staining reagent according to the manufacturer
표 BF inhibits the production of inflammatory cytokines in the BALF of OVA-induced asthma animal model. (A-C) The levels of IL-5, IL-6 and IL-13 were measured using ELISA assay. The absorbance was measured at 450 nm using microplate reader. Data are expressed as the mean ± S.D. #p< 0.01 indicates statistically significantly different from normal control group. **p < 0.01 indicates statistically significant difference compared to the OVA group
표 BF inhibits the production of IgE in the serum of OVA-induced asthma animal model. The levels of IgE were measured using ELISA assay. The absorbance was measured at 450 nm using microplate reader. Data are expressed as the mean ± S.D. #p< 0.01 indicates statistically significantly different from normal control group. p < 0.05 and **p < 0.01 indicates statistically significant difference compared to the OVA group
표 BF inhibits the influx of inflammatory cells and the production of MCP-1 in the lung and BALF of OVA-induced asthma animal model. (A-B) The levels of inflammatory cells influx and MCP-1 production were measured using H&E staining and ELISA assay. The absorbance was measured at 450 nm using microplate reader. Data are expressed as the mean ± S.D. #p< 0.01 indicates statistically significantly different from normal control group. *p < 0.05 indicates statistically significant difference compared to the OVA group
표 BF inhibits the overproduction of mucus in the lung of OVA-induced asthma animal model. The levels of mucus production were measured using PAS staining
표 꾸지나무 추출물의 UPLC-Q-TOF확인 조건
표 꾸지나무 추출물 및 유효분획물의 UPLC-Q-TOF-MS 크로마토그램
표 꾸지나무 추출물의 MPLC 크로마토그램
표 꾸지나무 뿌리에서 유효성분 20종 분리 모식도
표 꾸지나무 뿌리에서 분리된 유효성분들의 분광학적 데이터
표 꾸지나무 뿌리에서 분리된 BP 8의 분광학적 자료
표 꾸지나무 뿌리에서 분리된 BP 9의 분광학적 자료
표 꾸지나무 뿌리에서 분리된 BP 10의 분광학적 자료
표 꾸지나무 뿌리에서 분리된 BP 11의 분광학적 자료
표 꾸지나무 뿌리에서 분리된 BP 12의 분광학적 자료
표 꾸지나무 뿌리에서 분리된 BP 13의 분광학적 자료
표 꾸지나무 뿌리에서 분리된 BP 14의 분광학적 자료
표 꾸지나무 뿌리에서 분리된 BP 15의 분광학적 자료
표 꾸지나무 뿌리에서 분리된 BP 16의 분광학적 자료
표 꾸지나무 뿌리에서 분리된 BP 17의 분광학적 자료
표 꾸지나무 뿌리에서 분리된 BP 18의 분광학적 자료
표 꾸지나무 뿌리에서 분리된 BP 19의 분광학적 자료
표 꾸지나무 뿌리에서 분리된 BP 20의 분광학적 자료
표 대식세포에서 꾸지나무 추출물, 유효분획물, 분리된 13종 성분의 NO저해효과
표 대식세포에서 분리한 화합물 3종의 사이토카인 저해효과
표 대식세포에서 화합물 3종의 염증유전자 발현 억제효과
표 녹농균 감염된 H292 세포에서 MMP-12와 MMP-13의 증가
표 녹농균 감염된 H292 세포에서 NF-kB inhibitor 인 BAY11-7082의 MMP-12와 MMP-13 발현 억제 효과
표 급성폐렴 마우스 모델에서 녹농균에 의한 neutrophil 증가
표 녹농균에 특이적으로 감염된 A549 세포에서 Muc7, Muc13, Muc15, Muc19의 MOI 의존적 증가 mRNA level에서 real-time PCR 로 확인
표 녹농균에 특이적으로 감염된 A549 세포에서 Muc7, Muc13, Muc15, Muc19의 시간 의존적 증가 mRNA level에서 real-time PCR로 확인
표 녹농균에 감염된 A549 세포에서 Muc7, 13, 15, 19의 시간 의존적 증가 결과
표 급성 폐렴 마우스 모델에서 녹농균에 의한 염증 관련 세포 증가 확인
표 CuO에 의해 증가 및 BALF에서 CuO에 의해 neutrophil의 증가 확인
표 ExoS 특이적 반응을 이용한 ELISA assay system 원리
표 ExoS 특이적 반응을 이용한 ExoS-Flag ELISA assay system의 condition
표 ExoS-Flag ELISA assay로 성분은행 127개 시료의 ExoS 생성 억제활성 확인
표 ExoS-Flag ELISA assay에서 활성을 나타낸 31종 화합물의 H292에서의 세포생존율 측정
표 ExoS 억제활성 60% 및 세포생존율 80% 이상 22종 화합물들의 IL-1β 억제 활성
표 in vitro 에서 IL-1β 억제활성이 뛰어난 15종 화합물의 IL-1β 농도별 억제 활성 측정
표 The effect of NPFL10051.1 on type Ⅲsecretion system regulators and effector protein by quantitative PCR P. aeruginosa strains PAKΔST, PAKexoS were grown for 16h in LB broth on a rotary shaker at 37℃. The cultures were diluted 1: 20 in fresh LB broth supplemented with 1mM EGTA. The NPFL10051.1 was added to the broth at concentrations of 2.5-5-10-20 before incubation on a rotary shaker for 6h at 37℃
표 이번 연구에서 수행한 지방간과 당뇨의 치료적 타겟
표 황칠나무 잎 추출물의 TG 농도의존적 저해활성 결과
표 황칠나무 잎 추출물의 MPLC를 이용한 유효분획물
표 Lipid profile analyzed by TLC using either [14C] glycerol (A) or [14C] acetate (B) as radiolabeled substrates. Relative percentage of vehicle control represents was shown as a graph
표 Lipid profile analyzed by TLC using either [14C] glycerol (A) or [14C] acetate (B) as radiolabeled substrates. Relative percentage of vehicle control represents was shown as a graph
표 HOD에 의한 LKB1/AMPK 활성화 확인
$Inhibitory effect of total MeOH extract of DM-1 and its fractions on TAG biosynthesis related enzymes in vitro (enzyme source was obtained from hGPAT1, hDGAT1 overexpressed Sf-9 cells.). Final products measured by liquid scintillation counter (Tri-carb 2900TR Liquid Scintillation Analyzer; PerkinElmer)$ 표 Inhibitory effect of total MeOH extract of DM-1 and its fractions on TAG biosynthesis related enzymes in vitro (enzyme source was obtained from hGPAT1, hDGAT1 overexpressed Sf-9 cells.). Final products measured by liquid scintillation counter (Tri-carb 2900TR Liquid Scintillation Analyzer; PerkinElmer)
표 Effects of DM-1 on HFD-induced obesity in C57BL/6 mice. DM-1 (25 or 50 mpk) or orlistat (50 mpk) were gavaged orally administration. (A) Body weight in mice fed either a high fat or control diet for 8weeks. (B) A high fat diet significantly increases total fat, subcutaneous, epididymal and mesentery fat but DM-1 treatment group is reduced fat mass (C) Effect of DM-1 on serum leptin. Leptin was assayed by ELISA. (D) Liver weights of mice on high fat and control diet. (E) The level of plasma GOT and GPT. (F) Effect of DM-1 on serum lipid profile. (G) Steatosis in liver of high fat diet, DM-1 (250, 500 mpk) and orlistat (50 mpk) fed mice. The mice were sacrificed and liver tissue isolated. Histology was performed on H&E stained liver tissue as described
표 HepG2세포에서 TR 추출물의 농도의존적 중성지질 축적 억제 결과 (A) TR 잎 추출물 100 μg/ml 24시간 처리 후 Bodipy staining을 통해 HepG2 세포 내의 중성지방을 확인 함. (B) (A) TR 잎 추출물 50 μg/ml 24시간 처리 후 Nile red staining을 통해 HepG2 세포 내의 중성지방을 확인 함
표 Lipid profile analyzed by TLC using either [14C] acetate, [14C] glycerol, [14C] oleate as radiolabeled substrates. Relative percentage of vehicle control represents was shown as a graph
표 TR 추출물 처리 시 중성지방 생합성과정의 주요 전사인자인 SREBP-1과 그 하위 유전자들의 발현변화 양상
표 Inhibitory effect of spiramycin on lipid accumulation in 3T3-L1 adipocytes. (A) The effect of spiramycin on lipid droplet formation was measured by Oil Red O staining. 3T3-L1 preadipocytes were differentiated into adipocytes in the presence of various concentrations of spiramycin (2.5-20 μM) for 6 days. Representative cell images were captured at 200× magnification. GM, growth media. DM, differentiation media. (B) Quantification of intracellular lipid accumulation. Oil Red O stained lipids were extracted in absolute isopropanol, after which the absorbance of the solution was measured at 510 nm. (C) Quantification of triglyceride content. The bar graphs show the mean ± S.D. of 3 independent experiments(*p< 0.05, **p< 0.01, and ***p< 0.001 compared with the non-spiramycin-treated DM control)
표 Inhibitory effect of spiramycin on expression of adipogenic transcriptional factors and their adipocyte-specific target genes in 3T3-L1 adipocytes. (A) 3T3-L1 preadipocytes were differentiated into adipocytes in the presence of spiramycin for 6 days. The transcript levels of major adipogenic transcription factors (PPARγ, C/EBPα, and SREBP1) and their adipocyte-specific target genes (aP2, GLUT4, and FAS) were evaluated by qRT-PCR. (B) Western blotting analysis showing the effect of spiramycin on protein levels of major adipogenic transcription factors (PPARγ, C/EBPα, and SREBP1). The numbers at the bottom of the figure indicate the relative intensity of each band (fold-change in comparison with that of the control group), which was estimated using Multi Gauge software version 3.0
표 Effect of spiramycin on phosphorylation of AMPK and ACC during 3T3-L1 adipocyte differentiation. 3T3-L1 preadipocytes were differentiated into adipocytes with specified concentrations of spiramycin. After 1 h, protein levels of phosphorylated AMPK and ACC(p-AMPK and p-ACC) were analyzed by western blotting. The numbers at the bottom of the figure indicate the relative band intensity normalized to that of the non-phosphorylated protein (fold-change in comparison with that of the control group)
표 Effect of ZA-M on OA-induced intracellular lipid accumulation in HepG2 cells. (A) The cells were pre-treated with indicated concentrations of ZA-M for 1 h, then exposed to 500 mM OA for 24 h. Lipid accumulation was determined by using Oil Red-O and BODIPY493/503 staining. Nuclei were counterstained with Hoechst33342 dye. (B) Quantification of intracellular lipid accumulation. Oil Red O stained lipids were extracted in absolute isopropanol, after which the absorbance of the solution was measured at 510 nm. (C) Quantitative analysis was performed by flow cytometry after BODIPY493/503 staining. (D) Quantification of triglyceride content. The bar graphs show the mean ± S.D. of 3 independent experiments (*p< 0.05, **p< 0.01, and ***p< 0.001 compared with the DMSO control; *p< 0.05, **p< 0.01, and ***p< 0.001 compared with the OA treated control)
표 Effect of ZA-M on transcriptional/ translational expression of lipogenic genes and sequential TG biosynthesis in HepG2 cells. (A) Western blot analysis of lipogenic protein expression level was performed in HepG2 cells after treatment of indicated concentrations of ZA-M (0, 50, 30 and 10 ug/ml). (B) De novo TG biosynthesis. HepG2 cells were co-treated with various concentrations of ZA-M and [14C] glycerol for 6 h, and then TLC-based analysis of lipid intermediates was performed. Each [14C] TG bands was quantified by using a software, the Multi-Gauge V3.0 (Fujifilm). The relative activity was calculated by setting the value from DMSO-treated cells to 100%. The bar graphs show the mean ± S.D. of 3 independent experiments (*p< 0.05, **p< 0.01, and ***p< 0.001 compared with the DMSO control). (C and D) Effect of ZA-M on OA-induced upregulation of transcriptional/ translational expression of lipogenic genes was analyzed RT-PCR and western blot, respectively
표 ZA-M activates the LKB1/AMPK signaling pathway. (A) ZA-M stimulates the phosphorylation at Thr 172 of AMPKα and Ser 79 of its down-stream target ACC in HepG2 cells. (B) ZA-M reactivates the AMPK against OA-induced its dephosphorylation. (C) AMPK kinase activity. The bar graphs show the mean ± S.D. of 3 independent experiments (*p< 0.05, **p< 0.01, and ***p< 0.001 compared with the DMSO control; *p< 0.05, **p< 0.01, and ***p< 0.001 compared with the OA treated control). (D) ZA-M stimulates the phosphorylation at Ser 428 of LKB1
표 Effect of ZA-M on OA-induced ROS production. The cellular ROS level was measured by FACS using the H2DCFDA and HE probe. The numbers at the figure indicate the mean fluorescence intensity
표 Effect of ZA-M on OA-induced TNF-α production. (A and B) Expression of TNF- α was confirmed by RT-PCR and western blot, respectively. (C) The cell-free supernatants were collected and analyzed for TNF-α production by ELISA. The bar graphs show the mean ± S.D. of 3 independent experiments (*p< 0.05, **p< 0.01, and ***p< 0.001 compared with the DMSO control; *p< 0.05, **p< 0.01, and ***p< 0.001 compared with the OA treated control)
표 isotope-based glucose uptake assay 실험 원리
표 L6-GLUT4myc 세포에서 확인한 각 소재의 포도당 흡수율에 대한 EC50 계산
표 소재 간 병합처리 조건에 따른 포도당 흡수능 증가 확인
표 두 약제의 상가적 상호작용 평가를 위한 계산식
표 NMRC336과 HCA의 포도당 흡수능에 대한 EC50 산출
표 Isobologram analysis
표 NMRC-336이 분화된 L6-GLUT4myc의 세포생장과 포도당 흡수능에 미치는 영향
표 NMRC-336에 의한 GLUT4의 세포막 위치이동 확인
표 NMRC-336의 인슐린수용체 이하 AKT 경로 활성화 확인
표 NMRC-336의 AMPK 경로에 의한 포도당 흡수능 증가 기전

과제명(ProjectTitle) :	-
연구책임자(Manager) :	-
과제기간(DetailSeriesProject) :	-
총연구비 (DetailSeriesProject) :	-
키워드(keyword) :	-
과제수행기간(LeadAgency) :	-
연구목표(Goal) :	-
연구내용(Abstract) :	-
기대효과(Effect) :	-

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 제목(한글), 저자명(한글), 발행일자, 전자원문, 초록(한글), 초록(영문) 관리번호, 제목(한글), 제목(영문), 저자명(한글), 저자명(영문), 주관연구기관(한글), 주관연구기관(영문), 발행일자, 총페이지수, 주관부처명, 과제시작일, 보고서번호, 과제종료일, 주제분류, 키워드(한글), 전자원문, 키워드(영문), 입수제어번호, 초록(한글), 초록(영문), 목차
저장형식	Text(ASCII format) Excel format
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증

COPD/천식 등 만성질환 치료용 글로벌 천연물신약 연구사업
Development of natural drug candidates for the global level therapeutics against chronic inflammation related diseases such as COPD/asthma and obesity 원문보기

초록 ▼

Abstract ▼

목차 Contents

표/그림 (300)

표/그림 (300)

연구자의 다른 보고서 :

참고문헌 (25)

연구과제 타임라인

관련 콘텐츠

원문 보기

이 보고서와 함께 이용한 콘텐츠

연관된 기능

AI-Helper ※ AI-Helper는 오픈소스 모델을 사용합니다.

선택된 텍스트

연합인증

COPD/천식 등 만성질환 치료용 글로벌 천연물신약 연구사업 Development of natural drug candidates for the global level therapeutics against chronic inflammation related diseases such as COPD/asthma and obesity 원문보기

초록 ▼

Abstract ▼

목차 Contents

표/그림 (300) 모든 표/그림 보기

표/그림 (300) 슬라이드로 보기

연구자의 다른 보고서 :

오세량 (26) 이형규 (44) 안경섭 (30) 이현준 (3) 김종평 (12) 이수의 (3) 김문옥 (5) 류형원 (2) 김정희 (10) 권옥경 (8) 김두영 (11)

참고문헌 (25)

연구과제 타임라인

전체(0) 논문(0) 특허(0) 보고서(0)

전체(0) 논문(0) 특허(0) 보고서(0)

관련 콘텐츠

원문 보기

이 보고서와 함께 이용한 콘텐츠

연관된 기능

AI-Helper ※ AI-Helper는 오픈소스 모델을 사용합니다.

선택된 텍스트

COPD/천식 등 만성질환 치료용 글로벌 천연물신약 연구사업
Development of natural drug candidates for the global level therapeutics against chronic inflammation related diseases such as COPD/asthma and obesity 원문보기

표/그림 (300)

표/그림 (300)