Resveratrol에 의한 자궁근종세포의 성장억제 효과

강보영 ( Bo Young Kang ),조치흠 ( Chi Heum Cho ),김숙현 ( Sook Hyun Kim ),신소진 ( So Jin Shin ),권상훈 ( Sang Hoon Kwon ),박준철 ( Joon Cheol Park ),이정호 ( Jeong Ho Rhee ),김종인 ( Jong In Kim ),차순도 ( Soon Do Cha ),윤성도 ( 대한산부인과학회 2006 Obstetrics & Gynecology Science Vol.49 No.12

목적: 자궁근종세포에 resveratrol을 투여한 후 세포에 미치는 영향을 분석하여 치료에 유용한지를 알아보고자 하였다. 연구 방법: 자궁근종으로 수술 받은 10예의 환자에서 자궁근종과 정상 자궁근 세포를 배양하였다. Resveratrol의 자궁근종세포에 대한 세포생존율을 알아보기 위해 MTS 분석을 이용하였고, 세포 주기 회로에 관계하는 유전자의 발현을 보기 위해 Western blot analysis를 시행하였다. 세포주기분석은 FACS를 이용하였다. 결과: 일차 배양된 자궁근종세포에 resveratrol을 농도별로 처리한 결과 24시간 후 100 μM/L의 농도에서 30%의 증식억제 효과를 보였으며 농도가 증가할수록 증식억제 효과가 증가하였다. p53 단백은 농도가 증가할수록 발현이 증가하였고, resveratrol의 처리 농도에 비례하여 비활성 형태의 pro-caspase 3 단백질의 양적 감소와 PARP cleavage가 증가됨을 관찰하였고, pRB의 탈 인산화가 일어나는 것을 확인하였다. Resveratrol의 농도가 증가할수록 DNA 분절의 증가와, sub G1 주기의 지연 증가가 확인되어 세포자멸사의 양적 증거를 확인하였다. 결론: 이상으로 resveratrol이 자궁근종세포의 증식억제에 효과가 있으며, 이는 세포자멸사에 이르게 하고 세포주기와 세포자멸사에 관련된 유전자들의 발현에 영향을 미침으로써 향후 자궁근종의 치료나 예방에 있어서 효과적인 대체 치료 약물의 가능성이 있다고 사료된다. Objective: To examine the effect of resveratrol on cell proliferation and cell cycle progression in the human uterine leiomyoma cells. Methods: MTS [3-(4,5-dimethylthiazol-2-yl)5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] reduction assay was carried out to determine the viability of human uterine leiomyoma cells. Western blot analysis was done using anti pRB, anti- p21cip1/waf1, anti- p53, anti- cyclin E, anti CDK2 antibodies to detect the presence and expression of these proteins in treatment with resveratrol. DNA fragmentation assay was done to find the rate of apoptosis. Cell cycle analysis for resveratrol treated in human uterine leiomyoma cells was done by FACS (fluorescence-activated cell sorter) analysis. Results: Resveratrol induced growth inhibition in a dose dependent manner, treatment with 100 μM/L resveratrol blocked 30% cell growth. From Western blot analysis it revealed resveratrol induced the expression of p53 increasing. Caspase pathway was activated and cleavage of PARP was occurred. Apoptosis took place but in a reduced manner. FACS results showed that resveratrol increased the percentage of cells in sub G1 phase. Conclusion: Resveratrol, a dietry phytoalexin, inhibited cell proliferation and induced cell cycle arrest at sub G1 by enhancing the production of p53. These results indicate that resveratrol will be a promising agent chemopreventives or therapeutics against human uterine leiomyoma cells.

Resveratrol이 포도주의 관능평가에 미치는 영향

김태희(Tae-Hee Kim),이동희(Dong-Heui Yi),김형주(Hyung Joo Kim) 한국식품영양과학회 2009 한국식품영양과학회지 Vol.38 No.12

본 연구에서는 한국에서 시판되고 있는 14종의 포도주를 포도주 생산 연도별(2003, 2006, 2007), 포도 품종별(Malbec, Carmenere, Merlot, Syrah, Cabernet Sauvignon, Chardonnay), 포도주 생산 지역별(남아프리카 공화국, 아르헨티나, 호주, 미국, 칠레)로 준비하여 각 포도주의 resveratrol 함량을 HPLC로 분석하였다. 분석을 통하여 동일 회사, 동종 포도주라도 연도에 따라서 resveratrol 함량이 다르고, 같은 품종이라도 지역에 따라, 같은 자연환경이라도 품종에 따라 resveratrol 함량의 차이를 확인하고자 하였다. 그리고 포도주 생산 연도별(2003, 2006, 2007), 포도주를 제조할 때 사용한 포도 품종별(Malbec, Carmenere, Merlot, Syrah, Cabernet Sauvignon, Chardonnay), 포도주를 생산한 나라별(남아프리카 공화국, 아르헨티나, 호주, 미국, 칠레)로 관능평가를 실시하여 resveratrol 함량이 포도주에서 향(나무냄새, 페놀향), 맛(떫은맛), 색(붉은색, 황금색), 수렴성(쓴맛)과 무게감등에 영향을 주는지 알아보고자 하였다. 연구결과, 대부분의 적포도주는 resveratrol 평균 함량 0.2~5.8 ㎎/L 범주에 있었고, 백포도주는 대부분의 적포도주보다 낮은 함량으로 확인되었다. Resveratrol 함량의 분석 결과와 관능평가를 비교한 결과 phenol 화합물의 한 종류인 resveratrol 함량이 높을수록 관능평가의 점수도 평균적으로 높이 평가되었다. 14종의 포도주를 관능평가에서 백포도주는 대부분의 적포도주보다 모든 항목에서 낮은 점수를 얻었고, 적포도주는 껍질이 두껍고 씨가 굵고 많은 품종일수록 관능평가의 각 항목에서 평균적으로 높은 점수로 평가되었다. Resveratrol를 포함하는 포도주의 각종 phenol 화합물 성분은 포도껍질에 많이 존재하며, 포도주 제조 시 발효로 생성된 에탄올에 의하여 포도주에 용출되어, 적포도주는 백포도주보다 그 함량이 많고, 일반적으로 포도주는 1700~1900 ㎎/L의 농도를 가지고 있는데, 본 연구 결과 resveratrol 농도 비례관계에 있는 phenol계 물질들은 주로 적포도주에서 붉은 색깔과 씁쓸하고 텁텁한 맛의 평가에 영향을 주고, 포도주의 숙성에 많은 영향을 미칠 뿐 아니라 포도주의 향중 나무 냄새, 페놀 향에 영향을 주며, 수렴성과 무게감 등 관능평가에 긍정적인 영향을 주는 것으로 확인되었다. In this study, effect of resveratrol concentration on wine taste was investigated. Content of resveratrol in 14 different wines were quantified using HPLC. The resveratrol concentrations in the Australian wine samples (grape species: Syrah) with 3 vintage years were analyzed and different concentrations of resveratrol were observed (2.89~3.84 ㎎/L). Variation in wine grape species with the same wine manufacturer (Chile, 2006) and variation in different manufacturing country of origin with same wine grape species (Cabernet Sauvignon, 2006) also produced the variations in resveratrol concentration. The preference analysis (color, aroma, taste, astringency and overall acceptability) of the sample wines were determined by 15 panelists using 5-point hedonic scale. When the analysis results of the resveratrol concentrations in the sample wines were compared with the preference analysis of sample wines, the resveratrol concentration in the wine produced the positive effects in the wine preference analysis in the factors of aroma, taste and color.

Resveratrol Enhances 5-Hydroxytryptamine Type 3A Receptor-Mediated Ion Currents: The Role of Arginine 222 Residue in Pre-transmembrane Domain I

Lee, Byung-Hwan,Hwang, Sung-Hee,Choi, Sun-Hye,Shin, Tae-Joon,Kang, Jiyeon,Lee, Sang-Mok,Nah, Seung-Yeol Pharmaceutical Society of Japan 2011 Biological & pharmaceutical bulletin Vol.34 No.4

<P>Resveratrol, which is found in grapes, red wine, and berries, has many beneficial health effects, such as anti-cancer, neuro-protective, anti-inflammatory, and life-prolonging effects. However, the cellular mechanisms by which resveratrol acts are relatively unknown, especially in terms of possible regulation of receptors involved in synaptic transmission. 5-Hydroxytryptamine type 3A (5-HT<SUB>3A</SUB>) receptor is one of several ligand-gated ion channels involved in fast synaptic transmission. In the present study, we investigated the effect of resveratrol on mouse 5-HT<SUB>3A</SUB> receptor channel activity. 5-HT<SUB>3A</SUB> receptor was expressed in <I>Xenopus</I> oocytes, and the current was measured using a two-electrode voltage clamp technique. Treatment of resveratrol itself had no effect on the oocytes injected with H<SUB>2</SUB>O as well as on the oocytes injected with 5-HT<SUB>3A</SUB> receptor cRNA. In the oocytes injected with 5-HT<SUB>3A</SUB> receptor cRNA, co- or pre-treatment of resveratrol with 5-HT potentiated 5-HT-induced inward peak current (<I>I</I><SUB>5-HT</SUB>) with concentration-, reversible, and voltage-independent manners. The EC<SUB>50</SUB> of resveratrol was 28.0±2.4 μ<SMALL>M</SMALL>. The presence of resveratrol caused a leftward shift of 5-HT concentration–response curve. Protein kinase C (PKC) activator or inhibitor had no effect on resveratrol action on <I>I</I><SUB>5-HT</SUB>. Site-directed mutations of pre-transmembrane domain 1 (pre-TM1) such as R222A, R222D, R222E, R222K, and R222T abolished or attenuated resveratrol-induced enhancement of <I>I</I><SUB>5-HT</SUB>, indicating that resveratrol might interact with pre-TM1 of 5-HT<SUB>3A</SUB> receptor. These results indicate that resveratrol might regulate 5-HT<SUB>3A</SUB> receptor channel activity <I>via</I> interaction with the N-terminal domain and these results further show that resveratrol-mediated regulation of 5-HT<SUB>3A</SUB> receptor channel activity might be one of cellular mechanisms of resveratrol action.</P>

Resveratrol의 CD4+ T 세포 활성과 분화 억제 효과

서동원,이영주,이상명 한국자원식물학회 2014 한국자원식물학회지 Vol.27 No.5

Resveratrol is a naturally occurring stilbene which is safe and well-described compound with a potent anti-inflammatory activity. Recent studies suggested that resveratrol suppressed various inflammation mediated diseases such as asthma, chronic colitis, rheumatoid arthritis, and type 1 diabetes. These studies indicated that resveratrol might directly modulate CD4 + helper T cells (Th cells)-mediated immune responses. However, it is not fully elucidated whether resveratrol directly regulates CD4 + Th cell activation and differentiation. In the present study, CD4 + Th cells were purified from C57BL/6 and treated with various concentrations of resveratrol. We found that resveratrol directly suppressed CD4 + Th cells activation, leading to a defect in T cell proliferation. When CD4 + Th cells were treated with resveratrol, cytokine production was also significantly reduced in a dose dependent manner. In accordance with these results, resveratrol even inhibited CD4 + Th cells differentiation into Th1, Th2 or Th17, which produces IFN-γ, IL-4 or IL-17 respectively. We also found that resveratrol could induce apoptosis of CD4 + T cells at a high concentration. Our data demonstrated that resveratrol inhibited directly CD4 + Th cells activation and differentiation. It suggests that resveratrol could be an efficient therapeutic strategy for autoimmune diseases in which CD4 + Th cells play a critical role. Resveratrol은 천연 stilbene으로 안전성 있는 항염증 활성을 가진 화합물로 알려져 있다. 최근의 연구들에서 resveratrol이 천식, 만성 대장염, 류마티스성 관절염과 같이 염증에 의해발생하는 다양한 질병을 억제한다고 보고되었다. 이러한 연구들은 resveratrol이 CD4+ helper T cells (Th cells)에 의한 면역반응을 조절할 것이라고 제시하였다. 그러나 resveratrol이 직접적으로 Th cells의 활성화와 분화를 조절하는지 완전히 밝혀지지 않았다. C57BL/6에서 Th cells을 분리하여 다양한 농도의resveratrol을 세포에 처리하였다. 본 연구에서는 resveratrol이 직접적으로 Th cells의 활성화와 증식을 억제하는 것을 확인하였다. Th cells에 resveratrol을 처리하였을 때 IFN-γ, IL-4,IL-17 사이토카인 생성이 농도에 따라 유의하게 감소하였고 또한 Th cells이 이러한 사이토카인들을 분비하는 Th1과 Th2과Th17으로 분화되는 것이 억제되었다. 그리고 고농도의 resveratrol이 Th cells의 세포사멸을 유도하는 것으로 확인되었다. 본 연구에서는 resveratrol이 Th cells의 활성화와 분화를 직접적으로억제하는 것을 확인하였으며, 이는 resveratrol이 CD4+ Thcells에 의해 발생되는 자가면역질환의 효과적인 치료법이 될수 있을 것이라고 제시한다.

Primary 인체 전립선 암세포에서 Resveratrol의 Apoptosis 유도 효과

강혜인(Hye-In Kang),김재용(Jae-Yong Kim),조현동(Hyun-Dong Cho),박경욱(Kyung-Wuk Park),강점순(Jum-Soon Kang),서권일(Kwon-Il Seo) 한국식품영양과학회 2010 한국식품영양과학회지 Vol.39 No.8

본 연구에서는 resveratrol을 전립선 암 치료제로의 활용가능성을 조사하기 위하여 primary 인체 전립선 암세포에 대한 resveratrol의 성장억제 효과 및 그 기전에 대하여 조사하였다. Resveratrol은 RC-58T/h/SA#4 세포에서 농도 및 시간에 의존적으로 세포의 증식을 억제하였으며, IC50 값은 암세포인 RC-58T/h/SA#4, LNCaP, PC-3에서는 각각 245, 320, 340 μM, 전립선 정상세포인 RWPE-1에서는 982 μM로 나타나 정상세포에서보다는 암세포에서 그 독성이 크게 나타났다. 또한 resveratrol에 의해 유도된 세포 사멸은 핵 응축, sub-G1 함량 증가 및 DNA 분절 현상이 나타나 apoptosis를 유도함을 알 수 있었다. Resveatrol은 caspase-8, -9 및 effector casapse-3 활성을 농도 의존적으로 증가시켰으며, caspase 저해제인 z-VAD-fmk로 caspase의 처리 시resveratrol에 의한 apoptosis 유도 현상이 유의적으로 감소되어 resveratrol에 의한 RC-58T/h/SA#4 세포의 apoptosis 유도에 caspase가 중요한 역할을 하고 있음을 확인하였다. Resveratrol에 의해 anti-apoptotic 인자인 Bcl-2 및 Bid 단백질의 발현은 감소하였으나, pro-apoptotic 인자인 Bax 단백질 발현은 변화가 없었다. 따라서 본 연구는 resveratrol이 RC-58T/h/SA#4세포에서 caspase 의존형 미토콘드리아 경로에 의해 유도되며, resveratrol은 전립선암 치료제로서 사용 가능성을 시사한다. To evaluate resveratrol as a prostate cancer preventive material, we investigated its anti-proliferative and apoptotic effects in RC-58T/h/SA#4 primary human prostate cancer cells. Resveratrol significantly decreased the number of viable RC-58T/h/SA#4 cells in a dose- and time-dependent manner. Resveratrol showed cytotoxicity against RC-58T/h/SA#4, LNCaP, PC-3 human prostate cancer cells with IC50 values of 245, 320 and 340 μM, respectively. However the cytotoxic potential of resveratrol against normal RWPE-1 cells was lower (IC50=982 μM). Resveratrol induced cell death as evidenced by the increased formation of apoptotic bodies, nuclear condensation, sub-G1 phase, and DNA fragmentation. Resveratrol activated initiator caspases 8, and 9 as well as effector caspase 3 in a dose-dependent manner. Furthermore, the general caspase inhibitor z-VAD-fmk significantly inhibited resveratrol-induced apoptosis compared to cells without treatment. These results clearly indicate that resveratrol-induced apoptosis was dependent on caspase activation. Further, resveratrol modulated the down regulation of Bcl-2 (anti-apoptotic), and Bid. However, the level of Bax (pro-apoptotic) remained unchanged. These results suggest that resveratrol induced apoptosis in RC-58T/h/SA#4 cells via a mitochondrial-mediated caspase-dependent pathway, suggesting therapeutic potential against prostate cancer.

Inhibition of Sphingolipid Metabolism Enhances Resveratrol Chemotherapy in Human Gastric Cancer Cells

( Kyong Oh Shin ),( Nam Young Park ),( Cho Hee Seo ),( Seon Pyo Hong ),( Ki Wan Oh ),( Jin Tae Hong ),( Sang Kil Han ),( Yong Moon Lee ) 한국응용약물학회 2012 Biomolecules & Therapeutics(구 응용약물학회지) Vol.20 No.5

Resveratrol, a chemopreventive agent, is rapidly metabolized in the intestine and liver via glucuronidation. Thus, the pharmacokinetics of resveratrol limits its efficacy. To improve efficacy, the activity of resveratrol was investigated in the context of sphingolipid metabolism in human gastric cancer cells. Diverse sphingolipid metabolites, including dihydroceramides (DHCer), were tested for their ability to induce resveratrol cytotoxicity. Exposure to resveratrol (100 mM) for 24 hr induced cell death and cell cycle arrest in gastric cancer cells. Exposure to the combination of resveratrol and dimethylsphingosine (DMS) increased cytotoxicity, demonstrating that sphingolipid metabolites intensify resveratrol activity. Specifically, DHCer accumulated in a resveratrol concentration- dependent manner in SNU-1 and HT-29 cells, but not in SNU-668 cells. LC-MS/MS analysis showed that specific DHCer species containing C24:0, C16:0, C24:1, and C22:0 fatty acids chain were increased by up to 30-fold by resveratrol, indicating that resveratrol may partially inhibit DHCer desaturase. Indeed, resveratrol mildly inhibited DHCer desaturase activity compared to the specific inhibitor GT-11 or to retinamide (4-HPR); however, in SNU-1 cells resveratrol alone exhibited a typical cell cycle arrest pattern, which GT-11 did not alter, indicating that inhibition of DHCer desaturase is not essential to the cytotoxicity induced by the combination of resveratrol and sphingolipid metabolites. Resveratrol-induced p53 expression strongly correlated with the enhancement of cytotoxicity observed upon combination of resveratrol with DMS or 4-HPR. Taken together, these results show that DHCer accumulation is a novel lipid biomarker of resveratrol-induced cytotoxicity in human gastric cancer cells.

Resveratrol Up-regulates Cysteine-rich Angiogenic Inducer 61 (CYR61) in Human Colorectal Cancer Cells

Eun-Hee Kwak(곽은희),Jong-Sik Kim(김종식) 한국생명과학회 2013 생명과학회지 Vol.23 No.2

파이토케미칼 resveratrol은 항산화, 항염증, 항암등을 포함하는 다양한 생리활성을 가지고 있는 것으로 알려져 있다. 본 연구에서는 resveratrol이 CCN family 중의 하나인 cysteine-rich angiogenic inducer 61 (CYR61) 유전자의 발현을 유도할 수 있는지 연구하였다. 결과에 의하면 resveratrol은 3개의 다른 인간 대장암 세포주에서 CYR61 단백질의 발현을 유도하였을 뿐만 아니라, HCT116세포주에서는 처리한 resveratrol 농도와 시간 의존적으로 CYR61 단백질의 발현을 유도하였다. 이러한 CYR61 단백질의 발현이 resveratrol의 어떤 생리활성과 관련이 있는지 확인하기 위하여 몇 종류의 NSAIDs와 항산화제를 처리하여 CYR61 단백질의 발현을 확인하였으나, 오직 resveratrol의 처리에 의해서만 CYR61 단백질의 발현이 유도되었다. 또한, CYR61의 발현은 암 억제유전자인 p53과는 관련이 없는 것으로 판단되었다. Promoter assay를 통하여 프로모터 -732 ~ +54 사이에 조절부위가 있음을 확인하였고, 파이토케미칼 Indole-3-carbinol이나 6-gingerol에 의해서도 CYR61의 발현이 유도되지 않음을 확인하였다. 이러한 연구결과는 resveratrol에 의한 CYR61 유전자의 발현은 resveratrol특이적이며, 이러한 연구결과는 resveratrol만의 특이한 생리활성을 이해하는데 도움을 줄 것으로 기대된다. In this paper, we investigated whether resveratrol could induce the expression of cysteine-rich angiogenic inducer 61 (CYR61), which is a member of the CCN families. We showed that resveratrol up-regulated CYR61 protein expression in three different human colorectal cancer cell lines. In addition, resveratrol induced CYR61 protein expression in a dose- and time-dependent manner in a HCT116 cell line. To investigate the relationship between various biological activities of resveratrol and CYR61 expression, HCT116 cells were incubated with several NSAIDs, antioxidants, or resveratrol. Interestingly, resveratrol only induced CYR61 protein expression. The expression of CYR61 was not related to the presence of p53. A promoter assay revealed that the 786-bp promoter region (-732/+54) contains a regulatory region and that indole-3-carbinol and 6-gingerol could not induce CYR61 expression. In conclusion, our results indicate up-regulation of CYR61 is extremely resveratrol specific. The results can help to shed light on the unique biological function of resveratrol.

Resveratrol prevents alveolar bone loss in an experimental rat model of periodontitis

Bhattarai, G.,Poudel, S.B.,Kook, S.H.,Lee, J.C. Elsevier BV 2016 Acta Biomaterialia: structure-property-function re Vol.29 No.-

Resveratrol is an antioxidant and anti-inflammatory polyphenol. Periodontitis is induced by oral pathogens, where a systemic inflammatory response accompanied by oxidative stress is the major event initiating disease. We investigated how resveratrol modulates cellular responses and the mechanisms related to this modulation in lipopolysaccharide (LPS)-stimulated human gingival fibroblasts (hGFs). We also explored whether resveratrol protects rats against alveolar bone loss in an experimental periodontitis model. Periodontitis was induced around the first upper molar of the rats by applying ligature infused with LPS. Stimulating hGFs with 5μg/ml LPS augmented the expression of cyclooxygenase-2, matrix metalloproteinase (MMP)-2, MMP-9, and Toll-like receptor-4. LPS treatment also stimulated the production of reactive oxygen species (ROS) and the phosphorylation of several protein kinases in the cells. However, the expression of heme oxygenase-1 (HO-1) and nuclear factor-E2 related factor 2 (Nrf2) was inhibited by the addition of LPS. Resveratrol treatment almost completely inhibited all of these changes in LPS-stimulated cells. Specifically, resveratrol alone augmented HO-1 induction via Nrf2-mediated signaling. Histological and micro-CT analyses revealed that administration of resveratrol (5mg/kg body weight) improved ligature/LPS-mediated alveolar bone loss in rats. Resveratrol also attenuated the production of inflammation-related proteins, the formation of osteoclasts, and the production of circulating ROS in periodontitis rats. Furthermore, resveratrol suppressed LPS-mediated decreases in HO-1 and Nrf2 levels in the inflamed periodontal tissues. Collectively, our findings suggest that resveratrol protects rats from periodontitic tissue damage by inhibiting inflammatory responses and by stimulating antioxidant defense systems. Statement of significance: The aims of this study were to investigate how resveratrol modulates cellular responses and the mechanisms related to this modulation in lipopolysaccharide (LPS)-stimulated human gingival fibroblasts (hGFs) and protects rats against alveolar bone disruption in an experimental periodontitis model. Our findings suggest that resveratrol protects rats from periodontitic tissue damage by inhibiting inflammatory responses and by stimulating antioxidant defense systems. On the basis of our experiment studies, we proposed that resveratrol could be used as novel bioactive materials or therapeutic drug for the treatment of periodontitis or other inflammatory bone diseases like osteoporosis, arthritis etc. Furthermore, it could be also used for the modification or coating of implant materials as an antiinflammatory molecules which will help to accelerate bone formation. There are a few of reports suggesting antioxidant and anti-inflammatory potentials of resveratrol. However, our results highlight the cellular mechanisms by which resveratrol inhibits LPS-mediated cellular damages using human-originated gingival fibroblasts and also support the potential of resveratrol to suppress periodontitis-mediated tissue damages. We believe that the present findings might improve a clinical approach of using of resveratrol on human, although further detailed experiments will be needed.

Resveratrol이 근세포 내 AMPK 활성화와 미토콘드리아 생합성에 미치는 영향

정수련(SuRyunJung),김상현(SangHewunKim),김기진(KiJinKim) 한국체육학회 2013 한국체육학회지 Vol.52 No.2

본 연구의 목적은 resveratrol 처치가 근세포 내 AMPK 활성화와 미토콘드리아 생합성에 미치는 영향을 알아봄으로서 resveratrol이 ``exercise pill``로 활용될 수 있는지의 여부를 검증해보는 것이다. C2C12 mouse myoblast cell을 대상으로 농도에 따른 미토콘드리아 단백질과 AMPK 단백질의 변화양상을 관찰하기 위해 resveratrol의 최종 농도가 0,5, 10, 20, 50 uM이 되도록 처치하였으며, 1일 18시간, 총 3일간 처치한 후 harvesting하여 분석하였다. 연구 결과 resveratrol 처치는 C2C12 세포내 미토콘드리아 생합성을 증가시켰으며, AMPK, ACC의 인산화와 PGC-1α 단백질도 함께 증가시켰다. 그러나 세포독소를 측정한 결과 resveratrol 처치 후 24시간이 경과한 시점에서 20, 50 uM의 resveratrol 처치군이 0, 10 uM resveratrol 처치군에 비해 세포독소가 유의하게 증가한 것으로 나타났으며, 세포의 생존률도 유의하게 감소한 것으로 나타났다. 따라서 resveratrol 처치는 미토콘드리아 생합성을 증가시키나 이는 세포독성에 의한 ``인공적 효과``라고 생각된다. The purpose of this study to investigate the effects of resveratrol on AMPK activity and mitochondria biogenesis. C2C12 mouse myoblast cell were treated with 0, 5, 10, 20, 50 uM resveratrol (final concentration) for 16hr/day during 3 day. After treatment cells were harvested and analyzed. 3d resveratrol teat were significantly increase protein level of electron transfer chain enzyme and p-AMPK, pACC, PGC-1α. But at same concentration, Cytotoxicity was significantly increased and cell viability was significantly decreased. Therefore resveratrol increased mitochondria biogenesis bout this artificial effects induced by cytotoxicity.

Neuroprotective effects of resveratrol via anti-apoptosis on hypoxic-ischemic brain injury in neonatal rats

신진영,서민애,최은진,김진경,서억수,이준화,정혜리,김우택 대한소아청소년과학회 2008 Clinical and Experimental Pediatrics (CEP) Vol.51 No.10

Purpose : Resveratrol, extracted from red wine and grapes, has an anti-cancer effect, an antiinflammatory effect, and an antioxidative effect mainly in heart disease and also has neuroprotective effects in the adult animal model. No studies for neuroprotective effects during the neonatal periods have been reported. Therefore, we studied the neuroprotective effect of resveratrol on hypoxic-ischemic brain damage in neonatal rats via anti-apoptosis. Methods : Embryonic cortical neuronal cell culture of rat brain was performed using pregnant Sprague-Dawley (SD) rats at 18 days of gestation (E18) for the in vitro approach. We injured the cells with hypoxia and administered resveratrol (1, 10, and 30 µg/mL) to the cells at 30 minutes before hypoxic insults. In addition, unilateral carotid artery ligation with hypoxia was induced in 7-day-old neonatal rats for the in vivo approach. We injected resveratrol (30 mg/kg) intraperitoneally into animal models. Real-time PCR and Western blotting were performed to identify the neuroprotective effects of resveratrol through anti-apoptosis. Results : In the in vitro approach of hypoxia, the expression of Bax, caspase-3, and the ratio of Bax/Bcl-2, indicators of the level of apoptosis, were significantly increased in the hypoxia group compared to the normoxia group. In the case of the resveratrol-treated group, expression was significantly decreased compared to the hypoxia group. And the results in the in vivo approach were the same as in the in vitro approach. Conclusion : The present study demonstrates that resveratrol plays neuroprotective role in hypoxic-ischemic brain damage during neonatal periods through the mechanism of anti-apoptosis. Purpose : Resveratrol, extracted from red wine and grapes, has an anti-cancer effect, an antiinflammatory effect, and an antioxidative effect mainly in heart disease and also has neuroprotective effects in the adult animal model. No studies for neuroprotective effects during the neonatal periods have been reported. Therefore, we studied the neuroprotective effect of resveratrol on hypoxic-ischemic brain damage in neonatal rats via anti-apoptosis. Methods : Embryonic cortical neuronal cell culture of rat brain was performed using pregnant Sprague-Dawley (SD) rats at 18 days of gestation (E18) for the in vitro approach. We injured the cells with hypoxia and administered resveratrol (1, 10, and 30 µg/mL) to the cells at 30 minutes before hypoxic insults. In addition, unilateral carotid artery ligation with hypoxia was induced in 7-day-old neonatal rats for the in vivo approach. We injected resveratrol (30 mg/kg) intraperitoneally into animal models. Real-time PCR and Western blotting were performed to identify the neuroprotective effects of resveratrol through anti-apoptosis. Results : In the in vitro approach of hypoxia, the expression of Bax, caspase-3, and the ratio of Bax/Bcl-2, indicators of the level of apoptosis, were significantly increased in the hypoxia group compared to the normoxia group. In the case of the resveratrol-treated group, expression was significantly decreased compared to the hypoxia group. And the results in the in vivo approach were the same as in the in vitro approach. Conclusion : The present study demonstrates that resveratrol plays neuroprotective role in hypoxic-ischemic brain damage during neonatal periods through the mechanism of anti-apoptosis.