• 한국작물학회지
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• 제62권3호
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• pp.259-274
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• 2017

Platycodon grandiflorum (Bell flower) is an important plant that has traditionally been used as herbal medicine for the treatment of cough, phlegm, sore throats, lung abscesses, chest pains, dysuria, and dysentery. The present study was initiated to investigate the feasibility of inducing shoot and root organogenesis in cultured explants of P. grandiflorum in a range of culture media and through use of various plant growth regulators (PGRs). The plantlets (Stem containing one node) were isolated and cultured on different concentrations of Murashige and Skoog (MS) medium supplemented with PGRs. We found that proliferation and elongation of shoots and roots could be achieved on 1/4 MS for P. grandiflorum with wild and green petals and on 1/8 MS for P. grandiflorum with double petals. The highest levels of development and elongation of adventitious shoots and roots were observed when petal explants were cultured on 1/4 MS (pH 3.8) supplemented with 5% sucrose. Increasing the agar concentration reduced shoot growth and rooting potential; nevertheless, the highest number of shoots and roots was observed on 0.6% agar. In the case of growth regulators, 1/4 MS supplemented with $1mg\;L^{-1}$ 6-benzylaminopurine (BA) was found to be best for shooting, although higher concentrations of BA tended to reduce shoot and root elongation. The highest number of shoots was achieved on $0.5mg{\cdot}L^{-1}$ thidiazuron (TDZ) from double petal explants grown on 1/8 MS. However, root and shoot elongation were found to decrease when TDZ concentrations were increased. Low concentrations of kinetin, naphthalene acetic acid, indole acetic acid, and 3-indole butyric acid induced shoot and root proliferation and elongation. Taken together, our study showed that low concentrations of PGRs induced the greatest root formation and elongation, showing that the optimal concentration of PGRs for shoot proliferation was species-dependent.

• 한국식품과학회지
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• 제43권4호
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• pp.483-489
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• 2011

본 연구에서는 다양한 생리활성이 보고된 폴리페놀 화합물인 EGCG의 화학안정성, H2O2 생성능 및 세포독성에 대하여 다양한 항산화제와의 조합에 의한 변화를 분석하였다. EGCG는 생리적 조건에서 갈색화합물로 산화되면서 불안정화되는데, catalase를 제외한 각종 항산화제제 SOD, ascorbic acid, NAC 및 GSH는 EGCG 갈색산화물의 생성을 유의적으로 저해하였다. EGCG에 의해 생성되는 $H_2O_2$는 catalase에 의해 거의 완벽하게 제거되었으며, SOD와 NAC에 의해서도 유의적으로 감소하였다. 하지만 GSH 및 고농도의 ascorbic acid의 존재 시 오히려 $H_2O_2$ 수준이 증가하는 현상을 나타내었다. EGCG의 HeLa 및 HT-29 세포에 대한 독성은 catalase, SOD 및 NAC 등과 같은 항산화제 존재 하에 유의적으로 감소하였고 NAC에 의한 EGCG 세포독성의 감소는 첨가된 NAC의 농도 증가에 따라 더욱 두드러졌다. 그러나 GSH 존재하에 EGCG의 독성은 GSH와 EGCG농도에 따라 다른 조절 양상을 나타내었으며, ascorbic acid에 의해서 EGCG의 세포독성이 약간 증가하는 현상을 나타내었다. 본 결과는 EGCG와 함께 처리된 다양한 항산화제들이 ROS의 소거 뿐만 아니라 EGCG 화학안정성 등 다른 요인에 영향을 미칠 수 있으며, 항산화제의 존재 하에 변형된 EGCG활성에 대해 ROS관련 기작 외에 다양한 요인들에 대한 고려가 함께 이루어져야 함을 시사한다.

• 한국해양바이오학회지
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• 제2권2호
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• pp.102-107
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• 2007

산업의 발달과 함께 환경오염에 대한 국민적인 관심도는 날로 증가하고 있다. PCB는 우리 주변에 널리 퍼져 있고 먹이사슬을 통해 체내에도 축적되어 인체의 위해성이 우려되는 대표적인 환경오염물질이다. PCB의 노출은 성장기의 두뇌에서 가장 큰 신경독성을 나타내며 영아 및 유아는 상대적으로 높게 노출되어 위험집단으로 분류된다. 본 연구는 PCB의 신경독성에 구조-활성관계가 미치는 영향을 분석하고 PCB에 의한 독성을 최소화 할 수 있는 방안으로서 해양활성물질의 사용가능성을 이해하고자 하였다. PCB노출에 따른 신경세포의 신호전달 체계변화를 분석하기 위하여 Protein Kinase C (PKC)의 변화를 측정하였다. PKC의 전체적인 활성을 [$^3H$]PDBu로 분석한 결과 ortho-position(PCB-105, -123)을 가지고 있는 PCB가 non-ortho (pCB-77, -81) 구조보다 신경에 미치는 영향은 더 높았다. Westem blot 결과 PKC isofonn 중에는 PKC-beta II 및 epsilon의 경우 ortho-position PCB에서 더 높은 활성을 보였다. 이러한 PKC의 변화는 성장기 신경세포에서 신호전달기작의 변화에 많은 영향을 미치므로 이를 예방하거나 차단 할 수 있는 물질을 발견하고자 다양한 키토산을 처리하였다. 그 결과 1백만 달톤 이상의 고분자 키토산의 경우 PCB에 의한 신호전달 기작 변화를 억제할 수 있음을 보였다. 본 연구는 환경오염 등에 의한 독성예방에 키토산의 활용가능성을 제시하였다.

• 생명과학회지
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• 제28권5호
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• pp.569-576
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• 2018

본 연구에서는 흑색 방울토마토 주스의 항산화 활성, 아질산염 소거능, 알코올 분해능, 및 RAW 264.7 세포에서의 항염증 활성에 미치는 영향을 알아보았다. 흑색 방울토마토 주스의 총 페놀 함량은 $156.83{\mu}g\;TAE/ml$로 나타났다. 토마토주스의 항산화활성은 DPPH 라디칼 소거능과 SOD 유사활성으로 측정하였다. 토마토주스의 DPPH radical 소거능은 농도의존적으로 현저히 증가하였으며 40%의 농도에서의 83.39%로 나타났다. SOD 유사활성은 100% 농도에서의 SOD 유사활성은 22.01%로 나타났다. 흑색 방울토마토 주스의 숙취 해소능을 알아보기 위하여 체내 알코올 대사의 일차 효소인 ADH 활성 및 아세트알데히드를 분해하는 ALDH 활성을 알코올 분해와 숙취해소에 효과가 있는 것으로 알려진 hepos를 대조구로 하여 분석한 결과, ADH 활성 및 ALDH 활성 모두 농도 의존적으로 증가하였으며(p<0.05), 40% 농도에서의 ADH 활성과 ALDH 활성은 각각 198.87%와 181.89%로 높은 활성을 보였다. 아질산염 소거능 분석에서는 흑색 방울토마토 주스 50% 농도, pH 1.2, 3.0, 6.0에서 각각 85.06, 58.25, 43.68%로 pH가 낮을수록 아질산염 소거능이 증가하는 것으로 나타났다. 흑색 방울토마토 주스의 항염증 활성을 측정하기 위하여 LPS에 의해 유도된 RAW 264.7 대식세포의 NO 합성을 측정한 결과 농도 10%에서의 NO 합성은 1.28%로 LPS 처리군(7.79%)보다 83.57% 현저하게 감소하였다(p<0.05). 이상의 결과에서와 같이 흑색 방울토마토 주스는 항산화 활성과 알코올 분해능 및 항염증 효과가 높은 것으로 나타났기에 기능성 식품으로서의 가치가 높을 것으로 판단된다.

• 대한한의학방제학회지
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• 제19권1호
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• pp.207-217
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• 2011

Objectives : In a previous study, we have shown that Gagam-Gongjin-Dan(GGD) has an inhibitory effect on the ovalbumin-induced immune responses and a hepatoprotective effect on actaminophen-induced liver injury in Balb/c Mice. However, the possible anti-inflammatory effect of GGD extract for inflammatory mediators was not reported. Therefore, the purpose of this study was to investigate an inhibitory effects of GGD extract against lipopolysaccharides(LPS) induced inflammatory mediators in mouse peritoneal macrophages. Methods : GGD extract was prepared by extracting with methanol for 7 days. The extract was freeze-dried following filtration through vacuum distillation system. Accumulated nitrite, an oxidative product of nitric oxide(NO), was measured in the culture medium by the Griess reaction. The levels of prostaglandin $E_2(PGE_2)$, interleukin-$1{\beta}$(IL-$1{\beta}$), tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) were measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2(COX-2) were measured by Western blot analysis. Results : GGD extract (50-$400\;{\mu}g$/ml) per se had no cytotoxic effect in LPS-stimulated peritoneal macrophages. GGD extract dose-dependently reduced NO, $PGE_2$, IL-$1{\beta}$ and TNF-${\alpha}$ production and COX-2 activity caused by stimulation of LPS. The levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with GGD extract in a dose dependent manner. Conclusions : These results suggest that GGD extract has an anti-inflammatory effect against LPS-induced inflammatory mediators in peritoneal macrophages, these properties may contribute to inflammation disease care.

• Journal of Periodontal and Implant Science
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• 제32권4호
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• pp.733-744
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• 2002

The fibroblasts are the principal cells in the periodontal ligament of peridontium. As the periodontal ligament fibroblasts (PDLF) show similar phenotype with osteoblasts, the PDLF are thought to play an important role in alveolar bone remodeling. Cell-to-cell contacted signaling is crucial for osteoclast formation. Recently it has been reported that PDLJ enhance the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The aims of this study were to $clarify\;^{1)}$ the mechanism of PDLF-induced osteoclastogenesis $and\;^{2)}$ whether we can use preosteoclast cell line instead of primary hematopoietic preosteoclast cells for studying the mechanism of PDLF-induced osteoclastogenesis. Osteoclastic differentiation of mouse macrophage cell line RAW264.7 was compared with that of mouse bone marrow-derived M-CSF dependent cell (MDBM), a well-known hematopoietic preosteoclast model, by examining, 1) osteoclast-specific gene expression such as calcitonin receptor, M-CSF receptor (c-fms), cathepsin K, receptoractivator nuclear factor kappa B (RANK) ,2) generation of TRAP(+) multinucleated cells (MNCs), and 3) generation of resorption pit on the $OAAS^{TM}$ plate. RAW264.7 cultured in the medium containing of soluble osteoclast differentiation Factor (sODF) showed similar phenotype with MDBM-derived osteoclasts, those are mRNA expression pattern of osteoclast-specific genes, TRAP(+) MNCs generation, and bone resorbing abivity. Formation of resorption pits by osteoclastic MNCs differentiated from sODF-treated RAW264.7, was completely blocked by the addition of osteoprotegerin (OPG), a soluble decoy receptor for ODF, to the sODF-containing culture me야um. The effects of PDLF on differentiation of RAW264.7 into　the TRAP(+) multinucleated osteoclast-like cells were examined using coculture system. PDLF were fxed with paraformaldehyde, followed by coculture with RAW264.7, which induced formation of TRAP(+) MNCs in the absence of additional treatment of sODF. When compared with untreated and fixed PDLF (fPDLF), IL-1 ${\beta}$-treated, or lipopolysaccha-ride-treated and then fixed PDLF showed two-folld increase in the supporting activity of osteoclastogenesis from RAW264.7 coculture system. There were no TRAP(+) MNCs formation in coculture system of RAW264.7 with PDLF of no fixation. These findigs suggested that we can replace the primary hematopoietic preosteoclasts for RAW264. 7 cell line for studying the mechanism of PDLF-induced osteoclastogenesis, and we hypothesize that PDLF control osteoclastogenesis through ODF expression which might be enhanced by inflammatory signals.

• Journal of Periodontal and Implant Science
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• 제45권3호
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• pp.101-110
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• 2015

Purpose: Sclerostin, an inhibitor of Wnt/${\beta}$-catenin signaling, exerts negative effects on bone formation and contributes to periodontitis-induced alveolar bone loss. Recent studies have demonstrated that serum sclerostin levels are increased in diabetic patients and that sclerostin expression in alveolar bone is enhanced in a diabetic periodontitis model. However, the molecular mechanism of how sclerostin expression is enhanced in diabetic patients remains elusive. Therefore, in this study, the effect of hyperglycemia on the expression of sclerostin in osteoblast lineage cells was examined. Methods: C2C12 and MLO-Y4 cells were used in this study. In order to examine the effect of hyperglycemia, the glucose concentration in the culture medium was adjusted to a range of levels between 40 and 100 mM. Gene expression levels were examined by quantitative reverse transcription-polymerase chain reaction and Western blot assays. Top-Flash reporter was used to examine the transcriptional activity of the ${\beta}$-catenin/lymphoid enhanced factor/T-cell factor complex. Tumor necrosis factor-alpha ($TNF{\alpha}$) protein levels were examined with the enzyme-linked immunosorbent assay. The effect of reactive oxygen species on sclerostin expression was examined by treating cells with 1 mM $H_2O_2$ or 20 mM N-acetylcysteine. Results: The high glucose treatment increased the mRNA and protein levels of sclerostin. High glucose suppressed Wnt3a-induced Top-Flash reporter activity and the expression levels of osteoblast marker genes. High glucose increased reactive oxygen species production and $TNF{\alpha}$ expression levels. Treatment of cells with $H_2O_2$ also enhanced the expression levels of $TNF{\alpha}$ and sclerostin. In addition, N-acetylcysteine treatment or knockdown of $TNF{\alpha}$ attenuated high glucose-induced sclerostin expression. Conclusions: These results suggest that hyperglycemia increases sclerostin expression via the enhanced production of reactive oxygen species and $TNF{\alpha}$.

• 대한본초학회지
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• 제26권4호
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• pp.39-47
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• 2011

Objective: The roots, leaves, flowers, stems and seeds of Cirsium japonicum var. ussuriense are often used in treatment of human diseases such as hemorrhage, blood congestion and inflammation. Focusing our attention on natural and bioavailable sources of antioxidants and anti-inflammation, we undertook to investigate the antioxidant and anti-inflammatory properties of Cirsium japonicum var. ussuriense used as a folk medicine in Korea. Methods: The extracts of the leaves, stems, flowers, seeds and roots from C. japonicum var. ussuriense were prepared by extracting with water or 80% ethanol. Total flavonoids and polyphenols were measured by a colorimetric assay. The free radical scavenging activity of the extract was analyzed by the DPPH (1,1-diphenyl-2-picryl hydrazyl), ABTS (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) and Griess reagent assay. An oxidative product of nitric oxide (NO), was measured in the culture medium by the Griess reaction. The level of prostaglandin $E_2$ ($PGE_2$) was measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were measured by Western blot analysis. Results: Total flavonoid and polyphenol amounts of the leaves (CLE) and flowers (CFE) showed higher than those of the seed extract (CSE), stem extract (CSTE) and roots (CRE). CLE and CFE also showed the high antioxidant activities such as DPPH, NO-like and ABTS radical scavenging activity. An antioxidant activities of these water extracts showed higher than those of 80% ethanol extracts. We investigated the anti-inflammatory effects of CLE on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. CLE significantly suppressed the levels of the inflammatory mediators such as NO and prostaglandin $E_2$ ($PGE_2$) in dose dependant. Furthermore, the levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with CLE extract in a dose dependent manner. Conclusions: These results suggest that CLE water extract has a higher anoxidant and anti-inflammatory activity, these properties may contribute to the oxidative and inflammatory related disease care.

• 한국양식학회지
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• 제6권2호
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• pp.125-132
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• 1993

나일틸라피아의 전 암컷 생산을 유도하기 위하여 난황을 흡수하고 먹이를 먹기 시작하는 자어에 여성 홀몬인 $17\beta-estradiol$을 0, 60, 120, 240, 480ppm 농도로 먹이에 섞어 30일간 먹인 다음 성전환율, 생존율, 성장율 등을 조사하였다. 또한 이 홀몬 480ppm농도에서 투여 기간을 달리하여 10, 20, 30일간 먹인 효과도 조사하였다. 암컷의 출현 비율은 사료 중의 홀몬의 농도와 투여 기간에 비례 하였고 0, 60, 120, 240, 480 ppm 농도에서의 암컷 출현율은 각각 $47.5\%,\;86.4\%,\;91.3\%,\;97.0\%$ 및 $100\%$ 로 나타났으며, 480ppm에서 10, 20, 30 일간 먹 인 결과는 암컷 출현율이 각각 $64.2\%,\;84.3\%$, 및 $100\%$로 나타났다. 농도나 기간에 따른 생존율은 대조구와 차이가 없었으며 성장은 농도와 투여 기간에 비 례하여 낮게 나타났다. 따라서 이 종의 $17\beta-estradiol$에 의한 전 암컷 생산 가능 농도 및 기간은 480ppm으로 30일간 먹이는 것으로 나타났다.

• 동아시아식생활학회지
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• 제24권6호
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• pp.739-747
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• 2014

본 연구는 인체유방암세포인 MDA-MB-231세포를 이용하여 IDE를 처리하였을 때 세포사멸에 미치는 영향을 세포화학적 방법으로 확인하였다. IDE를 각각 0, 20, 30 및 $40{\mu}g/mL$ 첨가하여 24시간 배양한 후, 세포증식 억제, 막투과성, 세포내 ROS 분석 및 세포사멸 단계의 특성을 FACS 분석하고, RT-PCR에 의한 사멸관련 유전자 중 Bax/Bcl-2 ratio 분석을 통하여 IDE의 항암작용의 조절작용을 밝히고자 하였다. MTT의 세포 증식 억제는 첨가된 IDE의 농도에 따라 유의적으로 감소하였다(p<0.05). 이와 동시에, trypan blue에 대한 염색성과 DCF-DA 형광 분석의 결과는 각각, 막투과성과 세포내 ROS 농도가 모두 농도 의존적으로 증가됨을 보였다(p<0.05). 이와 같은 IDE 농도에 따른 세포화학적 변화 중에서 세포의 초기사멸에서 후기사멸 과정으로 급격한 사멸단계의 변화가 특히, IDE 농도 30과 $40{\mu}g/mL$에서 일어났다. RT-PCR 분석에 의한 Bax/Bcl-2 ratio도 IDE 농도 30과 $40{\mu}g/mL$에서 급격히 증가하였다(p<0.05). 이와 같은 세포화학적 결과와 RT-PCR 결과를 종합해 볼 때, IDE의 유방암세포(MDA-MB-231)에 대한 세포사멸작용은 막 투과성의 증가와 ROS 증가를 통하여 세포에 점진적인 손상을 일으키며, 이는 세포의 생화학적 변화도 초래하여 세포 증식 억제를 감소시켜, 결국 세포내의 사멸관련 유전자 지표인 Bax/Bcl-2 ratio를 크게 변화시키는 일련의 세포사멸을 점진적으로 유도시켜 항유방암의 효과의 가능성을 제시하였다.