• 대한수의학회지
• /
• 제39권2호
• /
• pp.301-310
• /
• 1999

Anthrax caused by Bacillus anthracis is one of the most important zoonotic diseases. The bacterium produces several virulence factors. Of the factors, protective antigen (PA) of tripatite toxin has been identified as a central component in the pathogenesis of anthrax. However, precise roles of PA and other cellular components in the reaction with the target cells remain to be elucidated, especially in the initial stage of the disease. Three B anthracis antigens were prepared for investigation; PA, sonicated cellular antigens (S-Ag) and formalin-inactivaed whole cell antigens (W-Ag). PA was purified from culture supernatant of the bacterium using FPLC system with MonoQ. S-Ag and W-Ag were prepared by sonication and formalin inactivation of the cultured cells, respectively. Purity of the antigens was confirmed by SDS-PAGE and Western blot analysis. The roles of these antigens in the production of inflammatory mediators such as NO, IL-6 and $TNF{\alpha}$ from mouse peritoneal macrophages were investigated. PA alone did not induce the production of the inflammatory mediators while the other antigens, S-Ag and W-Ag, did in a dose and time dependent manner. These results suggested that in addition to major virulence factors, other cellular antigens are also involved in the initial stage of the disease by the induction of inflammatory mediators.

• 대한화장품학회지
• /
• 제22권2호
• /
• pp.70-75
• /
• 1996

DMHF (2.5-dimethyl-4-hydroxy-3[2H]-furanone), an antioxidative compound from the reaction of L-cysteine/D-glucose scavenged efficiently 1,1-diphenyl-2-picryl hydrazyl free radicals. It exhibited an inhibitory effect on the autoxidation of linolenic acid, and the protective effect against UV cytotoxicity in cultured human fibroblast. In addition, DMHF appeared to prevent the cellular melanogenesis in the cultured murine melanoma cells more effectively than kojic acid, a well known inhibitor of melanogenesis, while the former was not so effective as the latter for the inhibition of the tyrosinase. Considering that cellular melanogenesis is a metabolic process triggered by oxidative stress, it ovas tentatively deduced that the antioxidative property of DMHF might afford the effect against cellular pigmentation by alleviating the causative stress. In toxicological tests such as irritation and sensitization, this compound turned out to be safe. The results of this study suggest that DMHF may be a novel inhibitor of melanogenesis, and that night be useful for application in cosmetics.

• Journal of Nutrition and Health
• /
• 제36권2호
• /
• pp.125-132
• /
• 2003

The present study was attempted to investigate and compare the antioxidant potency of several well-know flavonoids, antioxidant vitamin and commercially available popular beverages. The antioxidant potency was assessed by the effect on reducing oxidative DNA damage of human lymphocytes. Cellular oxidative DNA damage was measured by SCGE (single-cell gel electrophoresis), also known as comet assay. Lymphocytes were pre-treated for 30 minutes with wide ranges of doses of apigenin, kaempferol, luteolin, myricetin, rutin, quercetin, $\alpha$-tocopherol (10,25,50,100,200,500,1000 $\mu$M) ,green tea extract or grape juice (10,50,100,250,500,1000 $\mu$g/mL) followed by a $H_2O$$_2$(100 $\mu$M) treatment for 5 min as an oxidative stimulus. The physiological function of each antioxidant substance on oxidative DNA damage was analyzed as tail moment (tail length $\times$ percentage migrated DNA in tail) and expressed as relative DNA damage score after adjusting by the level of control treatment. Cells treated with $H_2O$$_2$alone (positive control) had an extensive DNA damage compared with cells treated with phosphate buffered saline (PBS, negative control) or pre-treated with all the tested samples. Of all the six flavonoids, quercetin was the most potent antioxidant showing the lowest $ED_{50}$/ of 8.5 $\mu$g/mL (concentration to produce 50% protection of relative DNA damage). The antoxidant potency of individual flavonoids were ranked as follows in a decreasing order; luteolin (18.4 $\mu$g/mL), myricetin (19.0 $\mu$g/mL) , rutin (22.2 $\mu$g/mL) , apigenin (24,3 $\mu$g/mL) , kaempferol (25.5 $\mu$g/mL). The protective effect of $\alpha$-tocopherol was substantially lower (highest $ED_{50}$value of 55.0 $\mu$g/mL) than all the other flavonoids, while the protective effect was highest in green tea and grape juice with low ED5O value of 7.6 and 5.3, respectively. These results suggest that flavonoids, especially quercetin, and natural compounds from food product, green tea and grape juice, produced powerful anti-oxidative activities, even stronger than $\alpha$-tocopherol. Taken together, supplementation of antioxidants to lymphocytes followed by oxidative stimulus inhibited damage to cellular DNA, supporting a protective effect against oxidative damage induced by reactive oxygen species.

• 한국응용과학기술학회지
• /
• 제32권4호
• /
• pp.781-788
• /
• 2015

본 연구에서는 건조된 까마중(Solanum nigrum L.) 전초의 50% 에탄올 추출물, 에틸아세테이트 분획, 아글리콘 분획을 이용하여 항산화 실험을 진행하였다. 1,1-diphenyl-2-picrylhydrazyl(DPPH)을 이용한 자유라디칼 소거 활성($FSC_{50}$)은 50% 에탄올 추출물, 에틸아세테이트 분획, 아글리콘 분획에서 각각 $215.46{\mu}g/mL$, $42.43{\mu}g/mL$, $52.28{\mu}g/mL$이였다. Luminol-의존성 화학발광법을 이용한 $Fe^{3+}-EDTA/H_2O_2$ 계에서의 총항산화능($OSC_{50}$) 평가에서, 까마중 50% 에탄올 추출물은 $25.25{\mu}g/mL$, 에틸아세테이트 분획은 $7.05{\mu}g/mL$, 아글리콘 분획은 $6.25{\mu}g/mL$를 나타냈다. $^1O_2$로 유도된 적혈구 세포손상에 대한 보호효과 측정에서는 까마중 50% 에탄올 추출물 및 아글리콘 분획은 $5{\sim}25{\mu}g/mL$ 농도에서는 세포보호효과를 나타냈으나 높은 농도에서는 세포보호 활성을 나타내지 않았다. 이러한 결과를 통해 까마중 추출물 및 분획물들은 항산화 활성을 가지고 있지만, 세포 수준에서는 비교적 높은 농도에서 활성산소로 유도된 세포 손상을 촉진할 수도 있음을 시사한다. 따라서 화장품 등에 까마중 추출물을 응용할 때는 사용상 주의가 필요한 것으로 판단된다.

• Preventive Nutrition and Food Science
• /
• 제16권1호
• /
• pp.12-17
• /
• 2011

This study was designed to investigate the protective effect of Sasa borealis leaf extract on 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress in LLC-PK1 cells (porcine kidney epithelial cells). The butanol fraction from Sasa borealis leaf extract (SBBF) was used in this study because it possessed strong antioxidant activity and high yield among fractions. Exposure of LLC-PK1 cells to 1 mM AAPH for 24 hr resulted in a significant decrease in cell viability, but SBBF treatment protected LLC-PK1 cells from AAPH-induced cell damage in a dose dependant manner. To determine the protective action of SBBF against AAPH-induced damage of LLC-PK1 cells, we measured the effects of SBBF on lipid peroxidation and antioxidant enzymes activities of AAPH treated cells as well as scavenging activities on superoxide anion radical and hydroxyl radical. SBBF had a protective effect against the AAPH-induced LLC-PK1 cellular damage and decreased lipid peroxidation and increased activities of antioxidant enzymes such as superoxide dismutase and glutathione peroxidase. Furthermore, SBBF showed strong scavenging activity against superoxide anion radical. The $IC_{50}$ value of SBBF was $28.45{\pm}1.28\;{\mu}g/mL$ for superoxide anion radical scavenging activity. The SBBF also had high hydroxyl radical scavenging activity ($IC_{50}=31.09{\pm}3.08\;{\mu}g/mL$). These results indicate that SBBF protects AAPH-induced LLC-PK1 cells damage by inhibiting lipid peroxidation, increasing antioxidant enzyme activities and scavenging free radicals.

• 한방비만학회지
• /
• 제20권1호
• /
• pp.10-19
• /
• 2020

Objectives: Oxidative stress-mediated neuroinflammation has been supposed as a crucial factor that contributes to the pathogenesis of many neurodegenerative diseases. In this study, we aimed to investigate the protective activity against oxidative stress and neuroinflammation of protocatechuic acid (PA), active phenolic compound from Momordica Charantia. Methods: Protective activity of PA from oxidative stress was performed under in vitro conditions. Our study investigated the protective mechanism of PA from neuroinflammation in cellular system using C6 glial cell. To investigate the improvement the effects on oxidative stress and neuroinflammation, we induced oxidative stress by H₂O₂ (100 μM) stimulation and induced neuroinflammation by treatment with lipopolysaccharide (LPS) (1 ㎍/mL) and interferon-gamma (IFN-γ) (10 ng/mL) in C6 glial cells. Results: PA showed strong radical scavenging effect against 1,1-dipenyl-2-picrylhydrazyl, hydroxy radical (·OH) and nitric oxide (NO). Under oxidative stress treated by H₂O₂, the result showed the increased mRNA expressions of oxidative stress markers such as nuclear factor-kappaB (NF-κB), cyclooxygenase (COX-2) and inducible nitric oxide (iNOS). However, the treatment of PA led to reduced mRNA expressions of NF-κB, COX-2 and iNOS. Moreover, PA attenuated the production of interleukin-6 and scavenged NO generated by both endotoxin LPS and IFN-γ together. Furthermore, it also reduced LPS and IFN-γ-induced mRNA expressions of iNOS and COX-2. Conclusions: In conclusion, our results collectively suggest that PA, phenolic compound of Momordica Charantia, could be a safe anti-oxidant and a promising anti-neuroinflammatory molecule for neurodegenerative diseases.

• Parasites, Hosts and Diseases
• /
• 제45권4호
• /
• pp.255-266
• /
• 2007

The protective effect of the Synadenium carinatum latex lectin (ScLL), and the possibility of using it as an adjuvant in murine model of vaccination against American cutaneous leishmaniasis, were evaluated. BALB/c mice were immunized with the lectin ScLL (10, 50, 100$[\mu}g$/animal) separately or in association with the soluble Leishmania amazonensis antigen (SLA). After a challenge infection with $10^6$ promastigotes, the injury progression was monitored weekly by measuring the footpad swelling for 10 weeks. ScLL appeared to be capable of conferring partial protection to the animals, being most evident when ScLL was used in concentrations of 50 and 100${\mu}g$/animal. Also the parasite load in the interior of macrophages showed significant reduction (61.7%) when compared to the control group. With regard to the cellular response, ScLL 50 and 100 ${\mu}g$/animal stimulated the delayed-type hypersensitivity (DTH) reaction significantly (P < 0.05) higher than SLA or SLA plus ScLL 10 weeks after the challenge infection. The detection of high levels of IgG2a and the expression of mRNA cytokines, such as IFN-$\gamma$, IL-12, and TNF-$\alpha$ (Th1 profiles), corroborated the protective role of this lectin against cutaneous leishmaniasis. This is the first report of the ScLL effect on leishmaniasis and shows a promising role for ScLL to be explored in other experimental models for treatment of leishmaniasis.

• Journal of Nutrition and Health
• /
• 제36권1호
• /
• pp.24-31
• /
• 2003

The present study was attempted to investigate the antioxidant capacity of popular yellow-green vegetable juices (kale, Angelica keishei, carrot, small water dropwort) and to investigate the effect of vegetable juices on protecting oxidative damage to DNA in cultured Chinese hamster lung (CHL) cells. Antioxidant capacity was analyzed by TRAP assay (Total radical-trapping antioxidant potential). Cellular DNA dmamage was measured by SCGE (single-cell gel electrophoresis, also known as comet assay. Cells incubated in medium with PBS (negative control) or with various concentration of the freeze dried green juices (25, 50, 100, 250 $\mu\textrm{g}$/$m\ell$) resuspended in PBS were treated with $H_2O_2$ (200 ${\mu}{\textrm}{m}$) as an oxidative stimulus for 5 min at 4$^{\circ}C$. The physiological function of each vegetable juice on oxidative DNA damage was analyzed and expressed as tail moment (tail length X percentage migrated DNA in tail) . Kale juice had the highest TRAP value suggesting that kale has the highest antioxidant capacity followed by Angelica keishei, small water dropwort and carrot. Cells treated with $H_2O_2$ had extensive DNA damage compared with cells treated with PBS or pre-treated with vegetable juice extracts. All green juices inhibited $H_2O_2$-induced DNA damage with kale being the most effective juice among the tested juices. These results indicate that green juice supplementation to CHL cells followed by oxidative stimulus inhibited damage to cellular DNA, supporting a protective effect against oxidative damage induced by reactive oxygen species. (Korean J Nutrition 36(1) : 24-31, 2003)

• 생약학회지
• /
• 제45권1호
• /
• pp.11-16
• /
• 2014

Protocatechuic acid is an active phenolic acid compound from Momordica charantia. In this study, we investigated the protective effect of protocatechuic acid against oxidative stress under cellular system using C6 glial cell. The oxidative stress was induced by hydrogen peroxide ($H_2O_2$) and amyloid beta 25-35 ($A{\beta}_{25-35}$), and they caused the decrease of cell viability and overproduction of reactive oxygen species (ROS). However, the treatment of protocatechuic acid significantly elevated the decreased cell viability and inhibited the overproduction of ROS by $H_2O_2$. In addition, protocatechuic acid significantly recovered the cellular damage induced by $A{\beta}_{25-35}$. In particular, protocatechuic acid at the concentration $10{\mu}g/mL$ decreased the elevated ROS level to normal level. These results indicate that protocatechuic acid may have neuroprotective effect through attenuating oxidative stress.

• Preventive Nutrition and Food Science
• /
• 제18권1호
• /
• pp.18-22
• /
• 2013

To investigate radical scavenging effects and protective activities of bitter melon (Momordica charantia) against oxidative stress, in vitro and a cellular system using LLC-$PK_1$ renal epithelial cells were used in this study. The butanol (BuOH) fraction of bitter melon scavenged 63.4% and 87.1% of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals at concentrations of 250 and $500{\mu}g/mL$, respectively. In addition, the BuOH fraction of bitter melon effectively scavenged hydroxyl radicals (${\cdot}OH$). At all concentrations tested, the scavenging activity of the BuOH fraction was more potent than that of the positive control, ascorbic acid. Furthermore, under the LLC-$PK_1$ cellular model, the cells showed a decline in viability and an increase in lipid peroxidation through oxidative stress induced by pyrogallol, a generator of superoxide anion ($O_2{^-}$). However, the BuOH fraction of bitter melon significantly and dose-dependently inhibited cytotoxicity. In addition, 3-morpholinosydnonimine (SIN-1), a generator of peroxynitrite ($ONOO^-$) formed by simultaneous releases of nitric oxide and $O_2{^-}$, caused cytotoxicity in the LLC-$PK_1$ cells while the BuOH fraction of bitter melon ameliorated oxidative damage induced by $ONOO^-$. These results indicate that BuOH fraction of bitter melon has protective activities against oxidative damage induced by free radicals.