• Title/Summary/Keyword: In-vitro study

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The Effect of TGF-{\beta}_1 on Cellular Activity of Periodontal Ligament Cells activated by PDGF-BB (PDGF-BB에 의한 치주인대세포활성에 대한 TGF-{\beta}의 효과)

  • Baek, Sang-Churl;Park, Jin-Woo;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.32 no.3
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    • pp.457-473
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    • 2002
  • The purposes of this study is to evaluate the combination effects of TGF-${\beta}_1$ and PDGF-BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/100% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Authors measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis according to the concentration of TGF-${\beta}_1$,(1,5ng/ml) and PDGF-BB (1,10 ng/ml) in combination. To explore further this delayed effect of TGF-${\beta}_1$, we preincubated human periodontal ligament cells with TGF-${\beta}_1$ for 4 or 24 hours before PDGF-BB stimulation. The results were as follows: The DNA synthetic activity was increased dose dependently by TGF-${\beta}_1$, PDGF-BB. The combination of TGF-${\beta}_1$ and PDGF-BB consistently enhanced the DNA synthetic activity to PDGF-BB alone. The ability of TGF-${\beta}_1$ to enhance DNA synthetic activity in PDGF-BB treated periodontal ligament cells was dose dependent. The maximum mitogenic effect was at the 5ng/ml of TGF-${\beta}_1$ and l0ng/ml of PDGF-BB. Preincubation of cell with TGF-${\beta}_1$ resulted in significantly greater response to PDGF-BB at all TGF-${\beta}_1$ concentration studied, and may be useful for clinical application in periodontal regenerative procedures. The total protein, collagen and noncollagen synthesis was increased dose pendently by TGF-${\beta}_1$, PDGF-BB. The % of collagen was slightly decreased according to the concentration of TGF-${\beta}_1$, PDGF-BB. The effect of TGF-${\beta}_1$, PDGF-BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. This study demonstrates that PDGF-BB is major mitogens for human periodontal ligament cells in vitro, and supports a role for TGF-${\beta}_1$ as a regulation of the mitogenic and total protein formation to PDGF-BB in these cells.

The Studies on Growth Characteristics and Dry Matter Yield of Hybrid Corn Varieties in Daegwallyeong Region (대관령에서 옥수수 품종별 생육특성과 건물수량에 기후변화의 영향)

  • Kim, Meing Jooung;Seo, Sung;Choi, Ki Choon;Kim, Jong Geun;Lee, Sang Hack;Jung, Jeong Sung;Yoon, Sei Hyung;Ji, Hee Chung;Kim, Myeong Hwa
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.33 no.2
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    • pp.123-130
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    • 2013
  • This study was conducted to investigate the effects of climate change on the growth characteristics and dry matter yields of silage corn hybrids in fields of forage crops of Hanwoo Experiment Station, National Institute of Animal Science, RDA, from Apr. 2009 to Sep. 2011. Corn hybrids were cultivated in Daegwallyeong of Gangwon Province, at an altitude of 760 m. Corn varieties used in this study consisted of 5 domestic varieties and 5 foreign varieties. Differences of silk days according to years occurred at an average of 5.5 days. The silk periods of domestic varieties occurred from Aug. 8 to 12, while that of overseas varieties was from Aug. 5 to 11. Silk days of domestic varieties occurred approximately 3 to 4 days earlier than those of oversea varieties. Silk days of Kwangpyeongok and P3156 belonging to the early varieties were Aug. 8 and 5, respectively. Kwangpyeongok and P3156 were Aug. 8 and 5, respectively. Pyeonganok and DK729 belonged to late varieties. The mean plant height of corn was approximately 231 cm, while those of Kwangpyeongok and Pyeonganok were 236 cm and 237 cm, respectively. The mean stem diameter and ear height of corn were approximately 23.2 mm and 94 cm, respectively. In the case of stem diameters, those of Cheonganok and 33J56 were 86 cm and 80 cm, respectively, while Gangdaok grew to a greater height (enter height) than other varieties. Dry matter yields of Kwangpyeongok and Pyeonganok were higher than those of other varieties. The mean leaf ratio of corn was 39.3%, while that of domestic varieties increased as compared to foreign varieties. The average DM yield of corn was 16,653 kg/ha, while those of 32P75, P3156, Pyeonganok, P3394 and Kwangpyeongok were 18,901, 17,997, 17,675, 17,194, 17,188 kg/ha, respectively. Total digestible nutrient (TDN) yields of 32P75, P3156, P3394, Pyeonganok and Kwangpyeongok were 13,381, 12,590, 12,532, 12,140 and 12,036 kg/ha, respectively. Corn crude protein (CP), in vitro dry matter digestibility (IVDMD), neutral detergent fiber (NDF), acid detergent fiber (ADF) and TDN were 7.8%, 74.2%, 42.4%, 23.5% and 70.3%, respectively. In the case of nutritive values of corn, there was no significant difference between of corn varieties of domestic and foreign origin.

Influence of Sodium Hypochorite & EDTA on the Microtensile Bond Strength of Ethanol Wet Bonding (Ethanol Wet Bonding에서 NaOCl과 EDTA가 결합강도에 미치는 영향)

  • Kim, Deok-Joong;Song, Yong-Beom;Park, Sang-Hee;Kim, Hyoung-Sun;Lee, Hye-Yoon;Yu, Mi-Kyung;Lee, Kwang-Won
    • Journal of Dental Rehabilitation and Applied Science
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    • v.29 no.1
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    • pp.37-44
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    • 2013
  • Sodium hypochlorite and ethylene diamine tetra acetic acid are substances usually used during endodontic treatment. Several studies found that the bonding was negated with certain irrigants and some of the used irrigants have demineralizing and chealating effects, so it was advocated to omit the etching step in etch and rinse adhesive systems. The purpose of this in vitro study was to evaluate the influence of NaOCl & EDTA on the bonding strength of ethanol wet bonding. Thirty human molars were selected and mesiodistally sectioned into halves, thus providing sixty specimens. The specimens were randomly assigned to 4 groups(n=15) according to the irrigant regimen used : (1) irrigated with distilled water for 10min (control); (2) irrigated with 5.25% NaOCl(10min), flushed with 5.25% NaOCl(1min) (3) irrigated with 5.25% NaOCl, flushed with 17% EDTA (4) irrigated with 5.25% NaOCl, flushed with 17% EDTA. Each group was acid-etched with 37% phosphoric acid(except group 4) and had their dentin surfaces dehydrated with ethanol solutions : 50%, 70%, 80%, 95%, 3x100%, 30s for each application. After dehydration, a primer( 50% all bond 3 resin + 50% ethanol) was used, followed by the adhesive(ALL-BOND 3 RESIN) application. Resin composite build-ups were then prepared using an incremental technique. Specimens were sectioned into beams and submitted to a tensile load using a Micro Tensile Tester(Bisco Inc.). The data were statistically analyzed using one-way ANOVA and Tukey HSD at p<0.5 level. There was no significant difference on G1(control) and G2(irrigated with NaOCl only ). (p>0.05). G3(flushed with EDTA) showed significantly high tensile bonding strength compared to the G2 (p<0.05). G4( treated with EDTA but no acid-etching) was significantly lower value than G3. (p<0.05) Although there was no significant difference, 5.25% NaOCl seemed to have an adverse effect on the bonding strength of ethanol wet bonding. The flushing with EDTA after NaOCl irrigation prevents the decrease of bonding strength. The use of 17% EDTA as a final flush can enhance the bonding strength but EDTA flushing can't substitute for a acid-etching.

Metabolic comparison between standard medicinal parts and their adventitious roots of Cynanchum wilfordii (Maxim.) Hemsl. using FT-IR spectroscopy after IBA and elicitor treatment (IBA 및 elicitor 처리에 따른 백수오 기내 생산 부정근 및 표준품의 FT-IR 스펙트럼 기반 대사체 비교 분석)

  • Ahn, Myung Suk;So, Eun Jin;Jie, Eun Yee;Choi, So Yeon;Park, Sang Un;Moon, Byeong Cheol;Kang, Young Min;Min, Sung Ran;Kim, Suk Weon
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.250-256
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    • 2018
  • To determine whether metabolite fingerprinting for whole cell extracts based on Fourier transform infrared spectroscopy (FT-IR) can be used to discriminate and compare metabolic equivalence, standard medicinal parts of Cynanchum wilfordii (Maxim.) Hemsl. and their adventitious roots were subjected to FT-IR. The principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) from FT-IR spectral data showed that whole metabolic pattern from the adventitious root of Cynanchum wilfordii was highly similar to its standard medicinal parts. These results clearly showed that mass proliferation of adventitious roots could be applied for the novel supply of standard medicinal parts of medicinal plants. Furthermore, FT-IR spectroscopy combined with multivariate analysis established in this study could be applied as an alternative tool for discriminating of whole metabolic equivalence from standard medicinal parts. Thus, it is proposed that these metabolic discrimination systems from the adventitious root of Cynanchum wilfordii could be applied for metabolic standardization of in vitro grown Cynanchum wilfordii.

Component Analysis and Antioxidant Activity of Oenanthe javanica Extracts (돌미나리 추출물의 함유성분 분석과 항산화 활성)

  • Hwang, Seok-Jun;Park, Sung-Jin;Kim, Jong-Dai
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.227-234
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    • 2013
  • The purpose of this study was to determine the possibility of using Oenanthe javanica as a natural health food source. To accomplish this, its general and biological activities were measured. Its carbohydrate, crude protein, crude lipid, and ash contents were 44.7, 9.8, 8.9, and 27.8%, respectively. The K content was largest for minerals followed by Ca, P, and Mg, which means that Oenanthe javanica is an alkali material. The concentrations of total phenol and flavonoids of OJE were $88.9{\pm}0.46$ mg GAE/g, and $28.6{\pm}0.64$ mg QE/g, respectively. Gallic acid, catechin, chlorogenic acid, and caffeic acid in OJE as measured by using HPLC were $0.9{\pm}0.23$, $1.2{\pm}0.19$, $227.1{\pm}0.62$, and $4.0{\pm}0.35$ mg/g. The DPPH and ABTS radical scavenging activities of OJE were 72.2%, and 66.1%, respectively, at $1,000{\mu}g/mL$. The FRAP and reducing power of OJE were 0.79, and 0.41 absorbance units value respectively, at $1,000{\mu}g/mL$. OJE possessed significant antioxidant properties, which suggests its great potential as a functional ingredient for food applications.

THE EFFECT OF WASHING PHOSPHORIC ACID ETCHANT ON SHEAR BOND STRENGTH OF AN ORTHODONTIC ADHESIVE (인산 부식액의 수세가 교정용 접착레진의 전단결합강도에 미치는 영향)

  • Kim, Hee-Kyun;Lee, Ki-Soo;Park, Young-Guk
    • The korean journal of orthodontics
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    • v.26 no.5 s.58
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    • pp.497-507
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    • 1996
  • The aim of present study in vitro was to evaluate and compare the effects of different washing times of enamels etched with low phosphoric acid solution which makes unsoluble salts and etched but contaminated with saliva on shear bond strength of an orthodontic adhesive to enamel, and to observe the washing effect on the etched enamel surface by scanning electron microscope. All brackets were bonded with Mono-$Lok2^{TM)}$) on the labial surface of extracted human bicuspids after etching with $20w/w\%\;and\;37w/w$ and phosphoric acid solution for 60seconds and then washing for 0,5,10 and 20seconds respectedly. After etching with $37w/w\%$ phosphoric acid solution and contaminating with saliva for 30seconds and then washing for 0,5,20 and 30seconds and re-etching for 10seconds. After 24hours passed in the $37^{\circ}C$ water bath, the shear bond strengths were measured on Universal Test Machine. The data were evaluated and tested by ANOVA and Duncan's multiple range test, and those results were as follows. 1. There was no significant differences between (p>0.05) shear bond strength of bonded brackets with 5, 10, 20seconds washing etched enamel using $37{\%}w/w{\%}$ phosphoric acid solution. 2. The shear bond strength of bonded brackets with $20w/w\%$ phosphoric acid and then washing for 5seconds showed bonded strength durable to occlusal force but its coefficiency score was high and etched surface was not cleaned completely and therefore it was assumed that its clinical application is not applicable. 3. There was no significant differences between (p>0.05) shear bond strengths of bonded brckets with washing for 5seconds etched enamel using $37w/w\%$ phosphoric acid solution and 10,20 seconds washing etched enamel using $20w/w\%$ phosphoric acid solution. 4. The shear bond strength of washing for 5seconds etched enamel which was contaminated with saliva showed sufficient bonded strength durable to occlusal force but its coefficiency score was high and therefore its clinical application was not applicable. 5. After etching, the sample contaminated with saliva showed the sufficient shear bond strength even washing 20seconds without re-etching.

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Elimination of Apple stem grooving virus from 'Mansoo' pear (Pyrus pyrifolia L.) by an antiviral agent combined with shoot tip culture (항바이러스제 처리와 경정배양에 의한 배(Pyrus pyrifolia L.) '만수'의 Apple stem grooving virus 무병화)

  • Cho, Kang Hee;Shin, Juhee;Kim, Dae-Hyun;Park, Seo Jun;Kim, Se Hee;Chun, Jae An;Kim, Mi Young;Han, Jeom Hwa;Lee, Han Chan
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.391-396
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    • 2016
  • In this study, in vitro-cultured 'Mansoo' pear (Pyrus pyrifolia L.) plants infected with Apple stem grooving virus (ASGV) were used for testing the efficiency of the virus elimination methods. The shoot tips cut from infected plants were treated by thermotherapy ($37^{\circ}C$), cold therapy ($4^{\circ}C$), chemotherapy with ribavirin, and combination of these methods. Treatment periods were 2, 4, and 8 weeks, and concentrations of ribavirin were 20 and $40mg{\cdot}L^{-1}$. The efficiency of ASGV elimination was evaluated by reverse transcription polymerase chain reaction. The shoot survival rate was the highest at 100% after cold therapy, chemotherapy, and combination of two methods, while the rate was the lowest at 33.3% after thermotherapy for 2 weeks. The shoot survival rate after chemotherapy decreased gradually as the treatment period was prolonged. The ASGV elimination rate was the highest at 100% after ribavirin treatment at a concentration of $40mg{\cdot}L^{-1}$ and combination of ribavirin treatment and thermotherapy for 2 weeks, whereas the ASGV elimination rate after cold therapy was the lowest at 16.7%. However, the efficiency of ASGV elimination was enhanced up to 43.3% by the combination of cold therapy and ribavirin treatment. The efficiency of ASGV elimination for all treatments was increased as the treatment period was prolonged. Based on these results, we suggest that ribavirin treatment at a concentration of $20mg{\cdot}L^{-1}$ for 4 weeks or at a concentration of $40mg{\cdot}L^{-1}$ for 2 weeks combined with shoot tip culture was efficient for the elimination of ASGV from pear.

Effect of Digestive Enzymes on the Allergenicity of Autoclaved Market Pork Sausages (가압가열 처리한 시판 돈육 소시지의 항원성에 미치는 소화효소의 영향)

  • Kim, Seo-Jin;Kim, Koth-Bong-Woo-Ri;Song, Eu-Jin;Lee, So-Young;Yoon, So-Young;Lee, So-Jeong;Lee, Chung-Jo;Ahn, Dong-Hyun
    • Food Science of Animal Resources
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    • v.29 no.2
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    • pp.238-244
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    • 2009
  • Food allergy is a serious nutritional problem in both children and adults. Therefore, food allergenicity reduction methods are greatly needed. The allergenicity is altered by various manufacturing processes, and the digestibility of food proteins can be affected by food processing. This study was conducted to investigate the effect of in-vitro digestibility on the allergenicity of autoclaved market pork sausages using pepsin (30min) and trypsin (5, 30, 60, 90, and 120min). The binding ability of the porcine serum albumin (PSA) from sausages A and B significantly decreased by about 30 and 23%, respectively, after autoclave treatment (121; 5, 10, and 30 min). After the pepsin and trypsin treatments, the binding ability of products A and B at 30 min decreased. These competitive indirect enzyme-linked immunosorbent assay (ci-ELISA) results corresponded well with the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting results. The results demonstrated that the allergenicity of pork sausages considerably decreased after autoclave treatment, and were also maintained or decreased after enzyme treatment. Accordingly, autoclave treatment represents a promising processing technology for the reduction of the allergenicity of diverse food products.

A Study on the In-Vitro Test of Thrombogenesis in Centrifugal Biopump (원심성 바이오 펌프를 이용한 혈전 생성의 생체 외 검사에 관한 연구)

  • Na, Myung-Hoon;Kim, Won-Gon;Kim, Joo-Hyun;Kim, Mee-Hyung;Yu, Jae-Hyeon;Lim, Seung-Pyung;Lee, Young
    • Journal of Chest Surgery
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    • v.32 no.10
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    • pp.863-873
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    • 1999
  • 배경: 혈전 생성은 심혈관계 삽입물이나 순환 보조장치의 발전에 있어서 중요한 문제이므로 이러한 장치들의 혈전성에 대한 평가는 필수적이다. 따라서 이러한 장치들의 혈전성 연구를 시행하기 위한 효과적인 생체 외 실험 방식이 개발되어 원심성 순환 펌프의 발전에 큰 도움이 되고 있으며, 최근 엄밀하게 혈액과 공기의 접촉을 차단하는 방식이 믿을 수 있는 생체 외 실험 방식으로 강조되고 있다. 본 논문에서는 모의 순환 회로를 이용하여 기존에 행해진 연구 방법 상의 공기 접촉에 따른 핼액 응고 기전의 활성화 여부를 알아보기 위하여 공기 접촉 여부에 따른 혈액 인자의 변화를 관찰하여 확인하고, 또한 회로 내에서 생성된 혈전을 생체 내에서 형성된 혈전과 비교 분석하여 이 모의 순환 회로를 이용한 생체 외 검사가 새로 고안된 심혈관계 장치들의 혈전성을 평가하는데 있어서 동물실험을 대신할 수 있다는 기존의 주장을 검증하고자 하였다. 연구재료 및 방법: 바이오펌프를 이용한 같은 모의 순환 회로 한 쌍을 준비하고 동일 개체에서 얻은 헤파린을 첨가한(1u/$m\ell$) 신선한 혈액을 공기와 접촉시키지 않도록 한 A-회로와 공기와 접촉시킨 B-회로에 충전시켜 실험을 동시에 진행하여 결과를 얻었으며 총 12번 시행하였다. 회로에 충전한 혈액의 activated clotting time(ACT)은 정상의 3~5배였으며 ACT가 정상 범위의 1.5배가되면 실험을 끝냈고, Hematocrit(HCT), 혈소판, 동맥혈가스분석(ABGA), factor Ⅷ, factor \ulcorner, 섬유소원, thromboxane B2(TXB2), free hemoglobin(fHb) 등을 측정하였다. 실험이 끝난 후 A-회로와 B-회로에서 얻은 각 검사를 평가하고 분석하였으며 생성된 혈전을 생체 내에서 생성된 혈전과 조직 검사로 비교하였다. 본 연구의 자료분석은 SPSS 통계 프로그램을 이용하였고 유의수준 0.05를 기준으로 유의도를 판단하였다. 결과: 정상 ACT는 186.9$\pm$20.5초(평균$\pm$표준편차)이었고, 헤파린을 넣은 초기 ACT는 982.2$\pm$165.5초이었으며, 실험시간은 보통 60분에서 180분 사이였다. HCT, fHb, 혈소판, TXB2, factor Ⅷ, factor \ulcorner, 섬유원소의 초기값은 35.5$\pm$3.2%, 12.4$\pm$6.5 mg/dL, 354$\pm$56($\times$103)/$\mu$L, 72.6$\pm$15.1 mg/dL, 29.3$\pm$1.2%, 137.9$\pm$42.1 mg/dL, 그리고 17.7$\pm$9.7%이었다. 공기 접촉에 따른 차이를 보았을 때 ACT(p=0.398), HCT(p=0.988), fHb(p=0.898), 혈소판 (p=0.904), TXB2(p=0.985), factor Ⅷ(p=0.872), factor \ulcorner(p=0.489), 섬유원소(p=0.973)으로 전 예에서 통계적 유의성이 없었다. 실험 후 양 군에서 생성된 혈전의 현미경 소견은 B-회로의 혈전에 여러 군데 공기 방울이 관찰되는 것 이외에 A-회로에서 생성된 혈전과 차이가 없었고, 생체에서 생성된 혈전과 비교했을 때 그 양상이 같았다. A-회로와 B-회로에서 생성된 혈전의 총 량은 8.4$\pm$3.7 mL와 9.4$\pm$3.1 mL로 통계상 유의성은 없었다. (p=0.624). 결론: 모의 순환 회로를 이용한 혈전성 평가를 위한 생체 외 실험에 있어서 공기와의 접촉 여부는 혈액 응고 기전의 활성화나 혈전 생성에 별다른 영향을 미치지 않는다. 모의 순환 회로에서 형성된 혈전과 문헌에 있는 인체 내에서 형성된 혈전을 비교한 결과 그 형태가 일치하므로 이 모의 순환회로를 이용한 실험이 동물실험을 대신할 수 있다고 판단된다.

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Nutritional Composition and Cytoprotective Effect of Moringa oleifera Lam. (Drumstick-tree (Moringa oleifera Lam.)의 주요 영양성분 및 추출물의 신경세포 보호 효과)

  • Jin, Su Il;Kim, Hyeon Ju;Jeong, Ji Hee;Jin, Dong Eun;Choi, Sung-Gil;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.46 no.5
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    • pp.609-615
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    • 2014
  • The cytoprotective effect of Moringa oleifera Lam. (drumstick tree) on neuronal cells was investigated to confirm the physiological benefits associated with this natural food resource. First, the drumstick tree extract was chemically analyzed to determine inherent nutritional constituents. Calcium and potassium were identified as the major mineral constituents, and palmitic acid (C16:0, 16.33%) and gadoleic acid (C20:01, 66.34%) were detected as the major fatty acids. Moreover, drumstick tree extract contained 94.78 mg/100 g vitamin E and 112.61 mg/100 g niacin. PC12 cells were used to study the cytoprotective effects of drumstick tree extract. Intracellular accumulation of reactive oxygen species was significantly reduced when $H_2O_2$ treated-neuronal cells were cultured in a medium containing the methanolic extract of drumstick tree, compared to cells treated with only $H_2O_2$. Cell viability assay using MTT showed that the extract protected cells against $H_2O_2$-induced neurotoxicity and inhibited LDH leakage from the cell membrane. Caspase assay showed that the extract exerted cytoprotective effect against apoptosis. Consequently, these data suggest that drumstick tree is a useful natural resource with positive effects on human health.