• 대한한의학방제학회지
• /
• 제25권4호
• /
• pp.509-526
• /
• 2017

Background and objectives : Soeumin Bojungykgi-tang (seBYTE) has been used to supplement qi in Korean medicine. It has been demonstrated to possess various biological functions such as anti-cancer, anti-aging and anti-inflammatory effects. The present study evaluated the protective roles of seBYTE in hepatotoxic in vitro and in vivo model. Methods : To investigate cytoprotective effect of seBYTE, HepG2 cells were pretreated with seBYTE and then subsequently exposed to $10{\mu}m$ AA for 12 h, followed by $5{\mu}m$ iron. Cell viability was examined by MTT assay, and expression of apoptosis-related proteins was evaluated by immunoblot analysis. For responsible molecular mechanisms, ROS production, GSH contents, and mitochondrial membrane potential were measured. In addition, hepatoprotective effect of seBYTE in vivo was assessed in $CCl_4$-induced animal model. Results : seBYTE prevented AA + iron-induced cytotoxicity in concentration dependent manner. In addition, ROS production, GSH depletion, and mitochondrial dysfunction induced by AA + iron were significantly reduced by seBYTE pretreatment. Furthermore, seBYTE recovered expression of the pro-apoptotic proteins such as PARP and pro-caspase-3. In animal experiment, plasma ALT and AST levels were significantly elevated in $CCl_4$ treatment, but seBYTE significantly decreased the ALT and AST levels. Moreover, seBYTE alleviated the numbers of histological activity index, percentages of degenerative regions, degenerated hepatocytes, infiltrated inflammatory cells, nitrotyrosine- and 4-hydroxynonenal-positive cells in liver. Conclusions : These results showed that hepatoprotective effect of seBYTE against on $CCl_4$-induced hepatic damages is partly due to antioxidative and anti-apoptotic process.

• Reproductive and Developmental Biology
• /
• 제33권2호
• /
• pp.99-105
• /
• 2009

Curcumin is a major active component of the food flovour tumeric. It has been used for the treatment of many diseases such as inflammatory and infectious diseases, cancer and other disease due to its antioxidant properties. Curcumin is a powerful scavenger of many free radicals such as superoxide anion, hydroxyl radical and nitric oxide. The objective of this study was to investigate the antioxidative effects of curcumin against hydrogen peroxide on semen quality during in vitro storage of boar semen. The sperm treated with different concentration of curcumin (1, 5 and 10 ${\mu}M$) in the presence or absence of hydrogen peroxide (250 ${\mu}M\;H_2O_2$) were incubated for 3, 6 and 9 hr at $37^{\circ}C$ and analyzed sperm characteristics such as motility, membrane integrity (MI), lipid peroxidation (LPO), reactive oxygen species (ROS) and DNA fragmentation (DF). The sperm motility and MI in $H_2O_2$ treated group ($47.8%{\pm}6.8$ and $24.8%{\pm}2.2$) were significantly decreased when compare to curcumin treated group ($79.8%{\pm}2.7$ and $34.6%{\pm}1.0$, respectively) irrespective of incubation periods(p<0.05). The LPO of spermatozoal plasma membrane was measured by thiobarbituric acid (TBA) reactions for malondialdehyde (MDA), MDA level in control ($11.6{\pm}0.6\;nmol/L{\times}10^6$) and curcumin groups ($10.7{\pm}0.3\;nmol/L{\times}10^6$) were lower than those of curcumin plus $H_2O_2$ ($17.1{\pm}0.8\;nmol/L{\times}10^6$) or $H_2O_2$ group ($22.5{\pm}1.9\;nmol/L{\times}10^6$) from 3 to 9 hr incubation periods. The DF by sperm chromatin dispersion (SCD) test and ROS production measured by 2',7'-dichlorofluorescein (DCF) fluorescence intensity were no significantly difference through all experimental groups (p>0.05). Correlation among evaluation methods for sperm quality, motility vs MI and DF vs ROS was positively correlated while motility vs DF and ROS vs LPO were negatively correlated in all treatment groups. These results demonstrate that curcumin can effectively improve the sperm quality during in vitro storage of boar semen through its hydrogen peroxide scavenging mechanism as an antioxidant.

• Nutrition Research and Practice
• /
• 제8권2호
• /
• pp.138-145
• /
• 2014

BACKGROUND/OBJECTIVES: This study was performed to investigate the in vitro antioxidant and cytoprotective effects of fermented sesame sauce (FSeS) against hydrogen peroxide ($H_2O_2$)-induced oxidative damage in renal proximal tubule LLC-PK1 cells. MATERIALS/METHODS: 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical ($^{\bullet}OH$), and $H_2O_2$ scavenging assay was used to evaluate the in vitro antioxidant activity of FSeS. To investigate the cytoprotective effect of FSeS against $H_2O_2$-induced oxidative damage in LLC-PK1 cells, the cellular levels of reactive oxygen species (ROS), lipid peroxidation, and endogenous antioxidant enzymes including catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-px) were measured. RESULTS: The ability of FSeS to scavenge DPPH, $^{\bullet}OH$ and $H_2O_2$ was greater than that of FSS and AHSS. FSeS also significantly inhibited $H_2O_2$-induced ($500{\mu}M$) oxidative damage in the LLC-PK1 cells compared to FSS and AHSS (P < 0.05). Following treatment with $100{\mu}g/mL$ of FSeS and FSS to prevent $H_2O_2$-induced oxidation, cell viability increased from 56.7% (control) to 83.7% and 75.6%, respectively. However, AHSS was not able to reduce $H_2O_2$-induced cell damage (viability of the AHSS-treated cells was 54.6%). FSeS more effectively suppressed $H_2O_2$-induced ROS generation and lipid peroxidation compared to FSS and AHSS (P < 0.05). Compared to the other sauces, FSeS also significantly increased cellular CAT, SOD, and GSH-px activities and mRNA expression (P < 0.05). CONCULUSIONS: These results from the present study suggest that FSeS is an effective radical scavenger and protects against $H_2O_2$-induced oxidative damage in LLC-PK1 cells by reducing ROS levels, inhibiting lipid peroxidation, and stimulating antioxidant enzyme activity.

• Journal of Nutrition and Health
• /
• 제42권8호
• /
• pp.750-758
• /
• 2009

건강한 성인 남자 48명을 대상으로 각각 당근즙 섭취군과 ${\beta}$-carotene 섭취군, placebo 약 섭취군으로 나누어 8주동안 실시하였으며, 당근즙은 매일 300 mL을 주었고 정제된 항산화 비타민은 당근즙에 함유된 ${\beta}$-carotene과 같은 양을 주어 항산화 영양 상태에 대한 개선 효과를 비교하고자 하였다. 그 결과, 혈장 비타민 C, 비타민 E 및 ${\beta}$-carotene 함량은 당근즙 및 항산화 비타민 섭취 전보다 유의적으로 높게 나타났고, 그 증가 정도가 비슷하게 나타나 식품으로부터 항산화 영양소를 충분히 섭취하는 것이 흡연자의 산화적 손상과 관련된 질병을 예방하는 좋은 방법이 될 수 있을 것이라 사료된다.

• 한국식품영양과학회지
• /
• 제41권1호
• /
• pp.7-13
• /
• 2012

본 연구에서는 산화적 스트레스에 의한 간 손상 개선 소재 개발을 위하여 꾸지뽕나무 각 부위별(잎, 줄기, 열매), 용매별(80% 에탄올, 10% 에탄올, 물) 추출물의 항산화 활성 및 간세포 보호효과를 측정하였다. 총 폴리페놀 함량과 플라보노이드 함량은 꾸지뽕 잎 80% 에탄올 추출물에서 가장 높게 나타났으며, 부위별로는 잎>줄기>열매 순이었고, 추출용매별로는 80% 에탄올>10% 에탄올>물 추출물 순으로 나타났다. DPPH 라디칼 소거능과 ABTS 라디칼 소거능 또한 잎80% 에탄올 추출물이 가장 높았으며, HepG2 세포에서 $H_2O_2$로 유도된 산화적 손상에 대해서는 꾸지뽕 잎 80% 에탄올 추출물만 유의적으로 높은 세포보호활성을 나타내었으며, HepG2/2E1 세포에서 알코올로 유도된 산화적 손상에 대한 각 부위별, 용매별 추출물의 간세포보호효과 또한 꾸지뽕잎 80% 에탄올 추출물이 가장 높게 나타났다. 부위별로는 잎>줄기>열매 순이었고, 추출 용매별로는 꾸지뽕 잎의 경우 80% 에탄올>10% 에탄올>물 순이었으며, 줄기와 열매의 경우는 용매별로 유의적인 차이가 나타나지 않았다. 이상의 결과로부터 꾸지뽕나무 잎 추출물은 우수한 항산화활성을 가질 뿐만 아니라 $H_2O_2$와 알코올로 유도된 간 손상으로부터 간세포 보호활성을 보임을 확인하였다. 이에 꾸지뽕나무 잎 추출물은 산화적 스트레스에 의한 간 손상으로부터 간세포 보호효과를 갖는 기능성 소재로 활용될 수 있을 것으로 사료된다.

• 식물조직배양학회지
• /
• 제26권1호
• /
• pp.53-58
• /
• 1999

Superoxide dismutase (SOD) 고생산세포주로 선발된 카사바 (Manihot esculenta Crantz) 배양세포에 감마선을 처리하여 세포생장과 SOD, peroxidase (POD), catalase (CAT)의 항산화효소 활성을 조사하였다. 카사바 캘러스는 계대배양 후 약 15일에 대수증식생장기에 진입한 후 30일에 최대생장을 나타내었으며 계속적인 배양에 따라 생장이 현저히 감소하였다. SOD와 POD의 비활성도 (units/mg protein)는 계대배양 직후에 가장 높았으며 대수증식 초기까지 감소한 후 다시 배양후기까지 증가하였다. CAT 활성은 세포생장과 비례하여 활성변화를 나타내었으며 계대배양 직후와 배양후기에 낮은 활성을 나타내었다. 계대배양 7일째의 카사바 캘러스에 감마선을 처리한 한 결과, 처리 후 14일째 배양세포는 방사선량에 비례하여 세포생장에 크게 영향을 주어 50Gy와 70Gy에서 각각 약 50%와 80%의 생장이 억제되었다. 이 시기의 SOD와 POD는 방사선량에 비례하여 증가하여 70Gy를 조사한 경우 각각 4배, 2.5배 증가하였으나 CAT은 방사선에 거의 영향을 받지 않았다. 카사바 캘러스배양에서 계대배양, 배양후기의 세포노화 및 배지내 영양고갈에 따른 배양스트레스는 SOD와 POD에 의해 주로 조절되고 있음이 시사되었다.

• 한국수정란이식학회지
• /
• 제28권3호
• /
• pp.243-250
• /
• 2013

Pyracantha is a genus of thorny evergreen large shrubs in the family of Rosaceae, with common names Firethorn or Pyracantha. It's extract has also been used in cosmetics as a skin-whitening agent and functioning through tyrosinase inhibition. Recent studies have shown that pyracantha extract possesses antioxidant activities and may significantly improve lipoprotein metabolism in rats. Although the mode of action of Pyracantha extract is not fully understood, a strong relationship was observed between antioxidant and apoptosis in some types of cells. Thus, the aim of this study was to evaluated the effect of pyracantha extract on blastocysts formation and their quality of the porcine parthenogenetic embryos. After parthenogenetic activation by chemicals, presumptive porcine parthenogenetic embryos were cultured in PZM-3 medium supplemented with extracts of pyracantha leaf, stalk and root for 6 day (1, 5 and $10{\mu}g/ml$, respectively). In our results, the frequency of blastocyst formation in pyracantha root extract ($5{\mu}g/ml$) treated group had increased that of other groups. Furthermore, blastocysts derived from pyracantha root extract ($5{\mu}g/ml$) treated group had increased the total cell numbers and reduced apoptotic index. Blastocyst development was significantly improved in the pyracantha root extract ($5{\mu}g/ml$) treated group when compared with the $H_2O_2$ treated group (p<0.05). Subsequent evaluation of the intracellular levels of ROS in pyracantha root extract ($5{\mu}g/ml$) treated groups under $H_2O_2$ induced oxidative stress were decreased (p<0.05). In conclusion, our results indicate that treatment of pyracantha root extract may improve in vitro development of porcine parthenogenetic embryos through its antioxidative and antiapoptotic effects.

• Asian-Australasian Journal of Animal Sciences
• /
• 제31권9호
• /
• pp.1420-1430
• /
• 2018

Objective: Epigallocatechin-3-gallate (EGCG) is a major ingredient of catechin polyphenols and is considered one of the most promising bioactive compounds in green tea because of its strong antioxidant properties. However, the protective role of EGCG in bovine oocyte in vitro maturation (IVM) has not been investigated. Therefore, we aimed to study the effects of EGCG on IVM of bovine oocytes. Methods: Bovine oocytes were treated with different concentrations of EGCG (0, 25, 50, 100, and $200{\mu}M$), and the nuclear and cytoplasmic maturation, cumulus cell expansion, intracellular reactive oxygen species (ROS) levels, total antioxidant capacity, the early apoptosis and the developmental competence of in vitro fertilized embryos were measured. The mRNA abundances of antioxidant genes (nuclear factor erythriod-2 related factor 2 [NRF2], superoxide dismutase 1 [SOD1], catalase [CAT], and glutathione peroxidase 4 [GPX4]) in matured bovine oocytes were also quantified. Results: Nuclear maturation which is characterized by first polar body extrusion, and cytoplasmic maturation characterized by peripheral and cortical distribution of cortical granules and homogeneous mitochondrial distribution were significantly improved in the $50{\mu}M$ EGCG-treated group compared with the control group. Adding $50{\mu}M$ EGCG to the maturation medium significantly increased the cumulus cell expansion index and upregulated the mRNA levels of cumulus cell expansion-related genes (hyaluronan synthase 2, tumor necrosis factor alpha induced protein 6, pentraxin 3, and prostaglandin 2). Both the intracellular ROS level and the early apoptotic rate of matured oocytes were significantly decreased in the $50{\mu}M$ EGCG group, and the total antioxidant ability was markedly enhanced. Additionally, both the cleavage and blastocyst rates were significantly higher in the $50{\mu}M$ EGCG-treated oocytes after in vitro fertilization than in the control oocytes. The mRNA abundance of NRF2, SOD1, CAT, and GPX4 were significantly increased in the $50{\mu}M$ EGCG-treated oocytes. Conclusion: In conclusion, $50{\mu}M$ EGCG can improve the bovine oocyte maturation, and the protective role of EGCG may be correlated with its antioxidative property.

• Nutrition Research and Practice
• /
• 제4권5호
• /
• pp.351-355
• /
• 2010

Our previous proteomic study demonstrated that oxidative stress and antioxidant delphinidin regulated the cellular level of $p27^{kip1}$ (referred to as p27) as well as some heat shock proteins in human colon cancer HT 29 cells. Current study was conducted to validate and confirm the regulation of these proteins using both in vitro and in vivo systems. The level of p27 was decreased by hydrogen peroxide in a dose-dependent manner in human colon carcinoma HCT 116 (p53-positive) cells while it was increased upon exposure to hydrogen peroxide in HT 29 (p53-negative) cells. However, high concentration of hydrogen peroxide (100 ${\mu}M)$ downregulated p27 in both cell lines, but delphindin, one of antioxidative anthocyanins, enhanced the level of p27 suppressed by 100 ${\mu}M$ hydrogen peroxide. ICR mice were injected with varying concentrations of hydrogen peroxide, delphinidin and both. Western blot analysis for the mouse large intestinal tissue showed that the expression of p27 was upregulated by 25 mg/kg BW hydrogen peroxide. To investigate the association of p27 regulation with hypoxia-inducible factor 1-beta (HIF-$1{\beta}$), the level of p27 was analyzed in wild-type mouse hepatoma hepa1c1c7 and Aryl Hydrocarbon Nuclear Translocator (arnt, HIF-$1{\beta}$)-defective mutant BPRc1 cells in the absence and presence of hydrogen peroxide and delphinidin. While the level of p27 was responsive to hydrogen peroxide and delphinidin, it remained unchanged in BPRc1, suggesting that the regulation of p27 requires functional HIF-$1{\beta}$. We also found that hydrogen peroxide and delphinidin affected PI3K/Akt/mTOR signaling pathway which is one of upstream regulators of HIFs. In conclusion, hydrogen peroxide and antioxidant delphinidin seem to regulate intracellular level of p27 through regulating HIF-1 level which is, in turn, governed by its upstream regulators comprising of PI3K/Akt/mTOR signaling pathway. The results should also encourage further study for the potential of p27 as a biomarker for intracellular oxidative or antioxidant status.

• 한국산학기술학회논문지
• /
• 제19권9호
• /
• pp.402-407
• /
• 2018

본 연구에서는 1.1-Diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical ($^{\cdot}OH$), 그리고 superoxide radical ($O_2{^-}$) 소거능 실험을 통해 쑥부쟁이의 항산화 효과를 평가하였다. 쑥부쟁이는 에탄올로 추출한 뒤, n-hexane, methylene chloride ($CH_2Cl_2$), ethylacetate (EtOAc), 그리고 n-butanol (n-BuOH)로 분획하였다. DPPH 라디칼 소거능 실험에서 모든 추출 및 분획물이 $10-100{\mu}g/mL$의 농도에서 농도 의존적으로 라디칼 소거능이 증가하였으며, 특히 EtOAc 분획물은 가장 강한 DPPH 라디칼 소거능을 나타내었다. 또한 쑥부쟁이는 $50{\mu}g/mL$ 농도에서 80% 이상의 $^{\cdot}OH$ 라디칼 소거율을 보였다. 특히 EtOAc 분획물은 $0.03{\mu}g/mL$에서 IC50 값을 나타내어 추출물과 분획물 중 가장 강한 $^{\cdot}OH$ 라디칼 소거능을 보여주었다. 또한, EtOAc 분획물은 $O_2{^-}$ 라디칼 소거능에서도 가장 강한 소거활성을 보였다. 이러한 결과는 쑥부쟁이가 산화스트레스로 인한 질병을 예방하는 천연 항산화제로서의 활용 가능성이 있음을 보여준다.