• The Korean Journal of Physiology and Pharmacology
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• 제24권1호
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• pp.11-18
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• 2020

The present study was aimed to explore the neuroprotective role of imatinib in global ischemia-reperfusion-induced cerebral injury along with possible mechanisms. Global ischemia was induced in mice by bilateral carotid artery occlusion for 20 min, which was followed by reperfusion for 24 h by restoring the blood flow to the brain. The extent of cerebral injury was assessed after 24 h of global ischemia by measuring the locomotor activity (actophotometer test), motor coordination (inclined beam walking test), neurological severity score, learning and memory (object recognition test) and cerebral infarction (triphenyl tetrazolium chloride stain). Ischemia-reperfusion injury produced significant cerebral infarction, impaired the behavioral parameters and decreased the expression of connexin 43 and phosphorylated signal transducer and activator of transcription 3 (p-STAT3) in the brain. A single dose administration of imatinib (20 and 40 mg/kg) attenuated ischemia-reperfusion-induced behavioral deficits and the extent of cerebral infarction along with the restoration of connexin 43 and p-STAT3 levels. However, administration of AG490, a selective Janus-activated kinase 2 (JAK2)/STAT3 inhibitor, abolished the neuroprotective actions of imatinib and decreased the expression of connexin 43 and p-STAT3. It is concluded that imatinib has the potential of attenuating global ischemia-reperfusion-induced cerebral injury, which may be possibly attributed to activation of JAK2/STAT3 signaling pathway along with the increase in the expression of connexin 43.

• Journal of Nutrition and Health
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• 제35권4호
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• pp.439-445
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• 2002

본 연구에서는 예로부터 민간과 한방에서 널리 이용되어 온 오매를 이용하여 오매 추출물이 간암 및 자궁암 세포주 증식에 미치는 효과를 알아보았다. 오매를 hexane. chloroform, ethylacetate, methanol 및 열수로 순차 추출하여 분획물을 농도별로 조제하였으며, MTT assay, cytotoxicity test 및 현미경 관찰 등을 통하여 암세포의 성장 억제 효과를 알아보았다. Hep G2의 경우 오매의 ethylacetate추출물이 가장 큰 성장 저해 효과를 나타내었으며 MTT assay결과 250 $\mu\textrm{g}$/ml 농도 이상에서부터 유의적인 증식 억제 효과를 보였다. 또한 cytotoxicity test결과 오매의 ethylacetate 추출물 1000$\mu\textrm{g}$/ml농도에서 72시간 배양시 75%의 세포 파괴율을 나타내었으며 현미경 관찰 결과 세포의 모양이 변화되고 죽은 세포가 배양액위에 떠있음을 관찰할 수 있었다. HeLa cell에서는 오매의 hexane 추출물이 가장 큰 저해 효과를 나타내었으며 MTT assay 결과 250 $\mu\textrm{g}$/ml 농도 이상에서부터 유의적인 증식 억제 효과를 보였다 또한 cytotoxicity test 결과 오매의 hexane추출물 1000 $\mu\textrm{g}$/ml농도에서 72시간 배양시 HeLa cell의 68%가 파괴되었으며 현미경 관찰 결과 세포 모양의 형태학적 변화가 일어났다. 정상세포주인 fibroblast에는 오매의 ethylacetate 및 hexane 추출물이 아무런 유의적 영향을 미치지 않았으며 현미경 관찰 시에도 변화가 일어나지 않았다

• Journal of Plant Biotechnology
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• 제39권3호
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• pp.163-168
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• 2012

본 실험은 TTC테스트를 통해 난과식물인 한라천마(G. verrucosa Blume)와 애기천마(H. sikokiana)의 종자기내배양시 NaOCl의 적정 처리시간과 농도조건을 확립한 결과이다. TTC테스트를 통한 종자활력은 애기천마의 경우 무처리구에서 95%이상 높았지만 배비대율, 비대배 직경과의 상관관계를 분석해보면 1%에서 30분간 처리 할 경우 종자활력대비 배비대율 77%, 배직경 $33{\mu}m$로 가장 높게 나타났다. 한라천마 역시 종자활력, 배비대율, 배직경 비대, 프로토콤 형성, 프로토콤직경 비대의 상관관계를 분석하면 1% NaOCl을 30분간 처리할 경우 최종적으로 종자활력대비 배비대율 87%, 배직경 $59{\mu}m$, 프로토콤의 형성율이 91%, 프로토콤 직경 $138{\mu}m$로 다른 처리구에 비해 높게 나타났다. 이상의 결과는 국내외적으로 연구된 바 없는 애기천마와 한라천마의 종자 기내배양 시 발아율을 개선하기 위한 중요한 기초자료로 활용될 것이고 더 나아가 자생 생물소재를 활용한 연구의 소재확보의 가능성을 제시하는 결과이다.

• Journal of Periodontal and Implant Science
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• 제31권1호
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• pp.149-165
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• 2001

This study was performed to define the cytotoxicity and the anti-inflammatory action of Pulsatilla koreana extracts. To analyze cytotoxic effects, gingival and periodontal ligament fibroblasts were used, and anti-inflammatory actions related to reduction of $IL-1{\beta}$ and $PGE_2$ production were performed in vitro, for the suggestion of efficacy and safety on periodontal therapeutic use of Pulsatilla koreana extracts. We extracted ethylacetate and butylalcohol from well-dried and ground Pulsatilla koreana throughout multiple processing, then used different concentration solution(0.1 %, 0.2 %, 0.4 %, 0.01 %, 0.02 %, 0.04 %, 1 %, 2 %) of ethylacetate and butylalcohol extracts to examine eytotoxic effects and anti-inflammatory actions Cytotoxic effects were examined by ELISA reader using MTT(Methyl Thiazol-2-YL-2, 5-diphenyl Tetrazolium bromide)solution following culture of human gingival and periodontal ligament fibroblasts. Synthesis of $IL-1{\beta}$was examined by $IL-1{\beta}$ enzyme-immunoassay(EIA)system after separation and culture of monocyte, and $PGE_2$ was examined by $PGE_2EIA$ system after culture of gingival fibroblasts. The results were as follows: 1. In the MTT test of gingival fibroblasts, the change of optical density was decreased significantly at 2 % of butylalcohol extracts and 0.04 %, 0.1 %, 0.2 %, 0.4 %, 1 %, 2 % of ethylacetate extracts.(p<0.05) 2. In the MTT test of periodontal ligament cells, the change of optical density were not differ significantly. but butylalcohol and ethylacetate extracts except from butylalcohol 0.01 % showed high cell cytotoxity. 3. Both ethylacetate and butylalcohol extracts from Pulsatilla koreana inhibited the synthesis of $IL-1{\beta}$and inhibition effect of ethylacetate extracts were higher than butylalcohol extracts. 4. Both ethylacetate and butylalcohol extracts from Pulsatilla koreana inhibited the synthesis of $PGE_2$, and ethylacetate extracts were higher than butylalcohol extracts. In conclusion, ethylacetate and butylalcohol extracts from Pulsatilla koreana showed little cell cytotoxity for gingival and periodontal ligament fibroblasts, and the inhibition of $IL-1{\beta}$ and $PGE_2$ sysnthesis, therefore it is considered that these extracts can be developed as the therapeutics of the periodontal disease.

• 한국식품과학회지
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• 제45권2호
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• pp.257-261
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• 2013

본 연구는 산화적 스트레스로부터 유도되는 신경세포 사멸을 보호할 수 있는 생리활성물질을 탐색하기 위하여 검정콩 껍질 추출물의 항산화 효과 및 PC12 신경세포 보호효과에 대하여 조사하였다. 다양한 용매에 의해 추출된 검정콩 껍질 추출물의 총 페놀화합물 함량을 측정한 결과 70% acetone 추출물이 다른 용매추출물에 비하여 가장 높은 총 페놀화합물 함량을 나타내었으며, 이를 기초로 70% acetone 추출물을 이용한 세포 내 항산화 활성도 높은 것으로 나타났다. 또한 PC12 신경세포 보호효과를 조사한 결과, 세포생존율과 세포막 손상 보호효과에서 70% acetone 추출물 처리구는 최대 $500{\mu}g/mL$까지 $H_2O_2$를 처리한 대조구에 비하여 농도 의존적으로 신경세포 보호효과가 나타남을 알 수 있었다. 동일 추출물을 활용한 in vivo 인지학습능력 평가에서도 대조구인 $A{\beta}$ ICV injection을 한 group과 비교하였을 때 전체적으로 추출물의 농도가 증가할수록 Y-maze test와 passive avoidance test에서 control구와 유사하거나 높은 인지 및 기억 능력 회복효과를 보여주었다. 본 연구결과를 종합해 보면 BSSCE는 항산화, 뇌 신경세포 보호효과 및 in vivo 인지 기억 회복능을 갖는 것으로 판단되며, 추후 검정콩 껍질에 존재하는 유효 생리활성물질을 확인함으로써 현대 사회의 가장 큰 질병 중 하나인 알츠하이머성 치매(AD)와 같은 퇴행성 뇌신경질환 예방에 유용한 소재가 될 것으로 판단된다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제13권4호
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• pp.391-404
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• 1991

Three anticncer agents which are different in time or dosage dependence as well as in phase specificity, namely mitomycin and adriamycin from natural products, and widely different cancer cell lines_Four epidermoid carcinomas originated from larynx, cervix, skin and gut were used toghether with one osteosarcoma as the target cell of single and combined administration of anticancer drugs. Semiautomated tetrazolium dye assay(MTT) appears to offer an attractive option for chemosensitivity of head and neck cancers since it is a simple, valid and inexpensive method of assessing chemosensitivity for large samples in a short time. The results obtained form this study were as follows. 1. Good correlations were obtained with the results of the MTT test and those of $^3H$ thymidine uptake assay. 2. $LD_{50}$ values of HIST and St.Ca. which showed relatively high doubling time on adriamycin were $30{\mu}g/ml$ and $15{\mu}g/ml$ while those of HeLa, Hep-2 and KHOS/NP were $2.1{\mu}g/ml$, $4.8{\mu}g/ml$, and $6.8{\mu}g/ml$ respectively. 3. The $LD_{50}$ value of 5-FU on five cancer cells were very high ranging from 15mg/ml to almost indefinite number, which means 5-FU is very resistant to epidermoid carcinomas or osteosarcoma examined in this study. 4. Mitomycin was relatively effective showing 80% cancer killing effect on HeLa, 70% on St. Ca. and 50% on Hep-2 at the high concentrations used. 5. Adriamycin was the most effective showing 90% cancer cell killing effect on KHOS/NP, 98% on HeLa, 80% both on Hep-2 and St. Ca. The least susceptible cancer cells toward adriamycin was HIST having only 55% cell killing effect at the high cincentration. 6. Combined therapy of adriamycin and 5-FU was more effective than single administration in all the cases examined. Most effective synergism was observed on St. Ca. at the low concentration, showing 21 times higher than each single administration.

• 대한예방한의학회지
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• 제21권3호
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• pp.87-98
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• 2017

Objectives : The objective of present study is to evaluate anti-arthritic effects of dried pomegranate concentrate powders (PCP), Eucommiae Cortex aqueous (EC) and ethanolic (ECe) extracts, Achyranthis Radix aqueous (AR) and ethanolic (ARe) extracts on the primary cultured rat articular chondrocytes. Methods : MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay was performed cytotoxic effect of test substances. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin $E_2\;(PGE_2)$ production and 5-lipoxygenase (LPO) activities, and inhibitory effects on metalloproteinase (MMP)-2 and MMP-9 activities were observed on the recombinant human interleukin $(rhIL)-1{\alpha}$ treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions - collagen type II, SOX9 and aggrecan. Results : As results, ECe and ARe showed obvious cytotoxicity against primary cultured rat articular chondrocytes at a dose level of 10 mg/ml, respectively. However, no obvious cytotoxic effects of PCP, EC and AR were demonstrated at a dose level of 10 mg/ml, on the primary cultured rat articular chondrocytes. In addition, treatment of LPS $50{\mu}g/ml$ induced significant increases of $PGE_2$ contents and 5-LPO activities indicating inflammatory responses of the primary cultured rat articular chondrocytes, and also decreases of cell viabilities, increases of MMP-2 and MMP-9 activities with decreases of extracellular matrix (ECM) related collagen type II, SOX9 and aggrecan mRNA expressions were observed by treatment of $rhIL-1{\alpha}$ 50 ng/ml, suggesting damages on the primary cultured rat articular chondrocytes and related ECM degradations. However, these inflammatory responses and related ECM degradations were inhibited by pretreatment of all test substances, in order of PCP > ECe > ARe > EC > AR, and $rhIL-1{\alpha}$ induced chondrocytes deaths are inhibited by treatment in order of PCP > EC > AR > ECe > ARe. Conclusions : Taken together, it is expected that mixed formulation of PCP as main components with appropriate proportion of EC and AR as additional components will be achieved a potent alternative medicinal food for osteoarthritis.

• Advances in Traditional Medicine
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• 제5권3호
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• pp.216-230
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• 2005

Although the field of study in immune enhancing compounds is relatively new, natural products from plants represent a rich and promising source of novel molecules with immunomodulating properties, Microglial cells, the main immune effector cells of the brain, usually display a ramified morphology and low expression levels of immunologically relevant antigens such as MHC class I and class II. Since any compound which participates in activation of phagocytic cells contributes to the production of potentially toxic factors, the search for convenient in vitro test-systems and study of mechanisms of action of these agents are of great interest. Human blood polymorphonuclear (PMN) cells and primary microglial cells isolated from Sprague-Dawley rats were used as cellular screening tests for study of phagocytosis-stimulating action of immunomodulating agents. Numbers of phagocytic activity were evaluated by the phagocyte ingestion of yeast cells and NO-synthase activity, nitrite production, and nitroblue tetrazolium test were determined after phagocyte stimulation. It was possible to demonstrate that indexes of phagocytic activity can be used as quantitative indicators for measurement immunomodulating activity. As a positive control, Zymosan A-induced phagocytosis in both PMN cells and primary microglial cells was used. $IFN-{\gamma}$ (0.1 -1 U/ml) stimulated phagocytosis in PMN cells 1.2 times after 2 - 3 h incubation, although at higher concentrations (10 - 100 U/ml) it strongly inhibited phagocytosis. In a similar way, at higher concentrations, $IFN-{\gamma}$ (100 - 500 U/ml) suppressed phagocytosis in zymosan-A stimulated microglial cells. When Polypodium leucotomus, cambricum and vulgare extracts were tested alone, increased levels of phagocytosis were observed in PMN. In addition, microglial cells showed both increased phagocytosis and MHC class-II antigen expressions. Surprisingly, when PMN and microglia were treated with a combination of Polypodium and $IFN-{\gamma}$, phagocytosis was not inhibited. We did not find changes in NO-synthase activity and nitrite production in both microglia and PMN cells activated by different immunomodulating agents. These results indicate that primary microglial cell cultures as well as human PMN cells can provide reproducible quantitative results in screening phagocytic activity of different immunoactive compounds. Furthermore, both inhibitory or activation mechanisms might be studied using these in vitro experimental approaches.

• 한국작물학회지
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• 제59권1호
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• pp.22-26
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• 2014

본 연구는 벼의 출수 후 일수에 따라 수발아 양상을 조사하고 수발아 피해가 발생할 수 있는 시기를 예측하여 수발아 방제를 위한 기초자료를 얻고자 수행하였으며, 그 결과를 요약하면 다음과 같다. 1. 벼 등숙기간 중 호품벼는 출수 후 20일, 신동진벼는 출수 후 30일부터 수발아 조건이 충족되면 발아가 시작되었으며, 50% 이상의 종자가 발아력을 갖는 시기는 호품벼가 약 35일, 신동진벼가 약 45일이었고, 출수 후 50일에는 두 품종에서 모두 80% 이상의 종자가 발아하였다. 2. 한 이삭 내에서 종자의 위치에 따른 수발아의 순서는 상위부 > 중위부 > 하위부 순으로 나타났으며 지경별로는 2차 지경보다는 1차 지경의 것이 높게 나타났다. 3. 수발아 검정 시 미발아된 종자는 배의 미숙, 종자휴면, 종자의 등숙불량 (불완전립)이 원인인 것으로 판단되었다. 4. 출수 후 30~45일의 등숙중기에 신동진벼는 호품벼보다 종자 휴면성이 높았다.

• 대한약침학회지
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• 제20권2호
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• pp.127-131
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• 2017

Objectives: Polymorphonuclear neutrophils (PMNs) constitute the first line of defense against invading microbial pathogens. Early events in inflammation involve the recruitment of neutrophils to the site of injury or damage where changes in intracellular calcium can cause the activation of pro-inflammatory mediators from neutrophils including superoxide generation, degranulation and release of myeloperoxidase (MPO), productions of interleukin (IL)-8 and tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$), and adhesion to the vascular endothelium. To address the anti-inflammatory role of flavonoids, in the present study, we investigated the effects of the flavonoids quercetin and vitexin on the stimulus-induced nitric oxide (NO), $TNF-{\alpha}$, and MPO productions in human neutrophils. Methods: Human peripheral blood neutrophils were isolated, and their viabilities were determined by using the Trypan Blue exclusion test. The polymorphonuclear leukocyte (PMNL) preparations contained more than 98% neutrophils as determined by morphological examination with Giemsa staining. The viabilities of cultured neutrophils with various concentrations of quercetin and vitexin ($1-100{\mu}M$) were studied using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without quercetin and vitexin ($25{\mu}M$) for 45 min, and stimulated with phorbol 12-myristate 13-acetate (PMA) ($10^{-7}M$). NO production was carried out through nitrite determination by using the Griess method. Also, the $TNF-{\alpha}$ and the MPO productions were measured using enzyme-linked immunosorbent assay (ELISA) kits and MPO assay kits. Results: Neutrophil viability was not affected up to a concentration of $100{\mu}M$ of quercetin or vitexin. Both quercetin and vitexin significantly inhibited $TNF-{\alpha}$, NO, and MPO productions in human neutrophils (P < 0.001). Conclusion:The present study showed that both quercetin and vitexin had significant anti-inflammatory effects. Thus, treatment with either quercetin or vitexin may be considered as a therapeutic strategy for treating patients with neutrophil-mediated inflammatory diseases.