난자의 성숙과정과 노화에 관한 이해는 인공수정과 체외수정 최적기를 판단하기 위하여 가장 중요한 연구내용으로 알려져 있다. 이러한 기작은 번식 호르몬들에 의하여 조절되는 것으로 알려져 있으나 난자 세포질 변화에 관한 내용은 잘 알려져 있지 않다. 본 연구에서는 산화질소물(nitric oxide, NO)이 난자 성숙과정에서 증가하는 것을 밝혔으며 난자의 미성숙단계(germinal vesicle stage, GV)와 난자핵막붕괴단계(germinal vesicle breakdown, GVBD) 및 성숙완료단계(metaphase II, MII)단계에서 생산되는 NO의 양을 비교하였다. 또한, 난자를 체외에서 배양할 때, MII단계로 성숙되지 않는 성장 단계의 난자에서는 NO의 증가 현상을 관찰할 수 없었고, 세포질이 불균일한 노화된 난자에서는 NO가 증가된 상태로 유지되는 특성이 있음을 밝혔다. 이러한 결과는 NO의 작용이 난자의 성숙과정과 난자 노화과정에서 중요한 기능을 담당하고 있음을 보여주고 있다.

Steaming is a method that has traditionally been used for medicinal plant extraction. This study investigated nitrite oxide production, ferrous ion chelating activity, α-glucosidase, xanthine oxidase, and acetylcholinesterase inhibitory activities of ethanol, acetone and hot-water extracts of Codonopsis lanceolata prepared by steaming seven times. MTT assay showed that each extract was non-toxic up to a concentration of 700 μg/mL confirming that there was no cytotoxicity in all extracts. The α-glucosidase, xanthine oxidase, and acetylcholinesterase inhibitory activities exhibited by the hot-water extract obtained from steaming seven times were higher (83.1%) than the other extracts. Higher production of nitrite oxide and better ferrous chelating activity was recorded with hot-water extract compared to ethanol and acetone extracts. These results indicated that more steaming of Codonopsis lanceolata extracts would be required to validate the possibility of developing antioxidants. Also, further study is needed to determine if the components present in the tested extracts might be useful in the prevention of Alzheimer's disease. These results showed that hot-water extracts may be useful for their antioxidant and the production inhibitory activity of nitrite oxide. It will be helpful in the investigation of the constituent analysis of the steam-processed product of Codonopsis lanceolata.

Colorectal cancer is a major cause of morbidity and mortality that accounts for over 9% of all incidences of cancer. Additionally, colorectal cancer is widely recognized as an environmental disease related to ill-defined cultural, social and lifestyle factors including physical activity, obesity, cigarette smoking and heavy alcohol consumption. Accordingly, natural phytochemicals and extracts have attracted attention because of their beneficial biological effects. Coenzyme Q10 (CoQ10) is a common supplementary medicine applied to increase bioenergetic capacity in various diseases. Therefore, in this study, we investigated whether CoQ10 treatment has any inhibitory effects and its related cellular mechanisms in human colon cancer HCT116 cells. A MTT assay revealed that CoQ10 slightly decreased the proliferation of HCT116 cells; however, glutathione- and superoxide dismutase- activity were unchanged in response to CoQ10 treatment. A DCF-DA assay revealed that CoQ10 slightly increased ROS release of HCT116 cells. However, in a nitric oxide (NO) assay, CoQ10 significantly increased NO production in a dose-dependent manner. The results of western blot analysis revealed that the protein levels of Bax, p21 and p53 were increased, whereas the protein level of Bcl2 was decreased suggesting that the CoQ10-mediated inhibitory mechanism is associated with apoptotic signaling. Taken together, our findings indicate that CoQ10 has an inhibitory effect on the growth of colon cancer cells via NO production that is associated with regulation of factors involved in apoptotic signaling including Bax, Bcl2, p21 and p53.

Russula rosacea, a mycorrhizal fungus, has been used for edible and medicinal purposes. This study was conducted to evaluate the in vitro antioxidant, anti-hyperglycemic, anti-cholinesterase, and nitric oxide inhibitory effects of the fruiting bodies from R. rosacea extracted with methanol, and hot water. The 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activities of the methanol and hot water extracts (2.0 mg/ml) of R. rosacea were comparable with BHT, the positive control. The chelating effects of the mushroom and hot water extracts were significantly higher than that of BHT. The reducing power of methanol and hot water extract (6 mg/ml) were significantly lower than that of BHT. Seven phenolic compounds were detected from acetonitrile and hydrochloric acid solvent extract of the mushroom. alpha-amylase and alpha-glucosidase inhibitory activities of methanol and hot water extracts were lower than that of acarbose, the positive control. The acetylcholinesterase and butyrylcholinesterase inhibitory effects were moderate compared with galanthamine, the standard drug. Nitric oxide (NO) production in lipopolysaccharide (LPS) induced RAW 264.7 cells were inhibited significantly by the mushroom extracts in a concentration dependent manner. Therefore, we demonstrated that fruiting bodies of R. rosacea possess in vitro antioxidant, anti-hyperglycemic, anti-cholinesterase, and NO production inhibitory activities. The experimental results suggest that the fruiting bodies of R. rosacea are good natural antioxidant, anti-hyperglycemic, anti-cholinesterase, and anti-inflammatory sources.

 ,  , Cordyceps (vegetable wasp and plant worm), an entomopathogenic fungi, has been used as a herbal medicine in Asian countries since ancient times. Cordyceps nutans is common but there is little research on this species. This study investigated the optimal culture conditions of C. nutans and the inhibitory effect on nitric oxide (NO) production in RAW 264.7 cell treated culture broth. The optimal conditions for the mycelial growth were 25℃ and pH 7.0-8.0. Mycelial growth was highest on mushroom complete medium (MCM), V8 juice agar (V8A), and yeast malt dextrose (YMD) medium. Mycelial growth on mushroom minimal medium (MMM) did not occur, so nutrient source was essential. Dextrose and sucrose as carbon sources, and ammonium citrate as a nitrogen source were satisfactory for mycelial growth. Cytotoxicity of C. nutans culture broth was not found in RAW 264.7 cells. C. nutans culture broth suppressed NO production of lipopolysaccharide (LPS)-stimulated RAW 264.7 cell in a dose-dependent manner. Thus, our results provided the optimal conditions for cultivation of C. nutans and showed that C. nutans may have excellent physiological activities.

In the present study, we have demonstrated that a novel synthetic chemical JSH-21 of N¹-Benzyl-4-methylbenzene-1,2-diamine could inhibit nitric oxide (NO) production in lipopolysaccharide (LPS)-activated macrophages RAW 264.7. The JSH-21 showed an IC50 value of 9.2 uM on the LPS-induced NO production. Furthermore, JSH-21 attenuated LPS-induced mRNA and protein levels of inducible NO synthase (iNOS) in the cells, as well as inhibited LPS-induced iNOS promoter activity. These results indicates hat the compound could down-regulate iNOS expression at the transcription level. Since NF-kB activation is a key mechanism in the expression of LPS-inducible iNOS gene, we further examined whether JSH-21 could affect LPS-induced NF-kB activation. JSH-21 inhibited LPS-induced nuclear import of NF-kB p65 in macrophages RAW 264.7 and sequentially prevented NF-kB transcriptional activity. However, JSH-21 was not effective in a LPS-induced degradation of cytoplasmic IkB-alpha in the cells. These results suggest that JSH-21 could inhibit LPS-induced NF-kB activation targeting nuclear import of NF-kB, an event downstream IkB degradation. Taken together, this study may provide a pharmacological potential of JSH-21 in the NO- or NF-kB-associated inflammatory disorders.

Nitric oxide(NO) is a labile, uncharged, reactive radical that functions as a sensitive mediator of intercellular communication in diverse tissues. It has been reported that NO is produced by osteoblast and these results may suggest that NO is integrally involved in the regulation of osteoclast formation and osteoclast resorption activity by osteoblastic cells. We examined the effect of cytokines on NO release by mouse bone marrow cell. We also examined the effects of cytokines and sodium nitroprusside(SNP) on the formation of osteoclast-like cell from mouse bone marrow cells in culture. Cytokines stimulated NO production of mouse bone marrow cells, and N-nitro-L-arginine methyl ester, a specific inhibitor of NO synthase, suppressed the cytokine-induced NO production. SNP showed dual action in the generation of osteoclasts. The addition of (30μM)SNP inhibited the formation of tartrate resistant acid phosphatase(TRAP)(+) multinucleated cell, whereas lower concentration(30μM) of SNP enhanced it. Although the precise action of NO remains to be elucidated in detail, the action of NO in osteoclast generation in our studies seems to be associated, at least in part, with bone metabolism and bone pathophysiology.

Inflammation in the brain has known to be associated with the development of a various neurologiacal diseases. The hallmark of neuro-inflammation is the activation of microglia, brain macrophage. Pro-inflammatory compounds including nitric oxide(NO) are the main cause of neuro-degenerative disease such as Alzheimer"s disease. In the study, we examined whether Harmonia axyridis extracts inhibit the NO production by a direct method using Griess reagent, western blotting and by RT-PCR(Reverse Transcription-Polymerase Chain Reactionin) the gene expression of inducible nitric oxide synthase(iNOS). Distilled water(H₂O) and methanol(MeOH) extracts of H. axyridis inhibited the protein expression of TNF-α(Tumor Necrosis Factor) and IL-6(Interleukin) in LPS (Lipo-polysaccharide) stimulated BV -2 cells at the concentration of 100 ng/㎖. Incubation of BV-2 cells with the extracts of H₂O of MeOH inhibited the LPS induced NO and iNOS protein. And this inhibition of iNOS protein is concordant with the inhibition of iNOS mRNA expression. These data suggested that H. axyridis extracts may play a crucial role in inhibiting the NO production.

만성적인 염증은 낭포성 섬유증, 암, 치매, 아토피성 질환, 비만 등과 같은 염증성 질환의 원인이 된다. 또한 염증의 발생단계에 관여하는 일부 신호물질은 피부조직의 손상과 노화에도 영향을 주는 것으로 알려져 있기 때문에 염증발생 매커니즘을 조절하기 위한 연구가 활발하게 이루어지고 있다. 최근에는 염증반응을 억제하거나 예방하기 위해, 몇몇 식물로부터 항염증 소재를 개발하려는 연구들이 이루어지고 있다. 특히 Stevia rebaudiana는 특유의 풍미를 가지는 천연감미료 스테비올배당체(steviol glycoside, SG)를 생성하는데, 일부 SG 에 대한 연구를 통해 염증억제 활성이 있는 것으로 알려져 있다. 본 연구에서는 기존연구를 통해 항염증 효능이 있는 것으로 확인된 스테비오사이드, 리버디오사이드 A, 스테비올 이외에도 항염증 소재로 활용될 가능성이 있는 SG가 더 존재할 것으로 예상하였다. 이를 확인하기 위하여 우리는 S. rebaudiana에서 얻은 SG의 nitric oxide (NO) 생성억제활성을 RAW 264.7 세포주를 대상으로 스크리닝 하였다. 그 결과 steviol β-glucopyranosyl ester (SGE)가 동일한 농도 조건의 SG 중에서 가장 높은 억제활성을 보여주었다. 또한, interleukin-1α (IL-1 α), interleukin-1β (IL-1β), cyclooxygenase-2 (COX-2), nuclear factor kappa-light chain-enhancer of activated B cells (NF-κB), inducible nitric oxide synthase (iNOS)와 같은 염증관련 인자의 mRNA 발현량 또한 농도의존적으로 감소시키는 것으로 확인되었다. 이러한 연구결과를 통해 SGE는 마우스 대식세포인 RAW 264.7 세포에서 항염증 활성 및 NO 생성 억제 효과가 있음을 확인하였다. 이를 통하여 SGE가 항염증 소재로 활용될 잠재성이 있음을 확인하였다.

Background: The flowering plant Hippophae rhamnoides L. has been used for many studies on fruit or leaf extracts. This study was conducted to investigate the development of a new cosmetic material from H. rhamnoides fruits and leaves that have by antioxidant, anti-inflammatory and wrinkle improvement activities. Methods and Results: The antioxidant abilities of H. rhamnoides extracts, including of a water-soluble fruit powder (FW), a fatsoluble fruit powder (FF), a supercritical extract of fruit by-product (BS), and a mixture of leaf and fruit (MIX), were investigated in vitro. A DPPH radical assay for antioxidant activity was performed for these fractions alongside assay to evaluate the total phenolic and flavonoid content (TPC and TFC). As expected, the MIX had the highest DPPH radical scavenging activity (RC50 = 10.27㎍/㎖), and the TPC and TFC also were highest in MIX (225.7 ㎎·GAE/g, and 25.18 ㎎·QE/g, respectively). Nitric oxide (NO) production in LPS-induced RAW264.7 cells was estimated and the results indicated an over 75% decrease of NO production in FF and MIX. In other assays, the highest elastase inhibitory activity was found in FW. Conclusions: These results revealed that H. rhamnoides extracts have a high potential for antioxidant, anti-inflammatory and antiwrinkle activities. H. rhamnoides products are suggested to be applied as the functional materials of cosmetic ingredients.

Background: Inula japonica Thunb. is a plant belonging to the family compositae. Inulae flos (flower of I. britannica var. chinensis Regal.) is the dried flower of I. japonica Thunb. and contains various flavonoids (patulitrin, nepitrin and kaempferol), which have been utilized in traditional oriental medicine to treat nausea, phlegm, and coughs. However, ethanol extract of I. britannica (IJE) has not been previously studied for its use in cancer treatment, and its effects on oxidative stress, or inflammation. Thus, the present study investigated the anti-oxidant, anti-inflammatory, and anti-colorectal cancer effects of IJE using RAW264.7 and HCT- 116 cells, which are human colorectal cancer cell line. Methods and Results: IJE contained flavonoids (80.95 ± 5.3 ㎎/g) and polyphenols (310.53 ± 10.6 ㎎/g). Moreover, it reduced lipopolysaccharide (LPS)-induced nitric oxide (NO) production and H2O2-induced oxidative stress by decreasing reactive oxygen species (ROS) levels. Additionally, the 500 ㎍/㎖ IJE treatment increased caspase-3 activity and apoptotic cell death in HCT-116 cells. Conclusions: These results demonstrate that the anti-cancer effect of IJE against human colorectal cancer cells involves caspase-3 activation and apoptotic cell death. IJE also inhibited LPS-induced NO production, and H2O2-induced oxidative stress in RAW264.7 cells. However, further studies are required to explore how IJE treatment regulates signal transduction in NO and ROS production.

해방풍이 가지는 건강 기능성 소재로서의 가치를 확인하고자, 해방풍 부위별 용매추출물의 항산화 활성 및 면역조절 효과를 조사하였다. 해방풍 부위별 용매추출물의 추출수율은 10.73-30.57이며, 총 폴리페놀 및 총 플라보노이드 함량은 해방풍 잎 70% 에탄올 추출물(EL)에서 각각 10.79 g/100 g 및 2.01 g/100 g으로 가장 높은 값을 나타내었다. DPPH radical 및 ABTS radical 소거활성은 해방풍 잎 70% 에탄올 추출물(EL)이 100-1,000 μg/mL 농도에서 12.90-84.70% 및 11.78-57.64%로 가장 우수한 radical 소거활성을 나타내었으며, superoxide radical 소거활성 및 FRAP 활성은 해방풍 잎 70% 에탄올 추출물(EL)이 100-1,000 μg/mL 농도에서 각각 49.91-84.05% 및 255.38-975.28 μM로 가장 우수한 활성을 나타내었다. 대식세포주인 RAW 264.7 세포에 해방풍 부위별 용매추출물을 처리하였을 때 nitric oxide 생성 억제능을 확인하기 위해 분석한 결과, 추출용매에 따른 nitric oxide 생성량은 70% 에탄올 및 70% 메탄올 추출물에서 가장 낮은 생성량을 보였으며, 해방풍 부위별에 따른 nitric oxide 생성량은 뿌리, 잎 및 씨앗 순으로 낮았다. 그 중에서 해방풍 뿌리 70% 에탄올 추출물(ER)에서 가장 낮은 nitric oxide 생성량을 보여 우수한 nitric oxide 억제 효과를 확인하였으며, 이를 이용하여 기능성 소재, 미용식품 및 화장품 제조용 원료 등 다양하게 활용 가능할 것으로 사료된다.

Background: To date, the anti-wrinkle efficacy of phellodendri cortex has not been defined. In this study, we investigated the nitric oxide (NO) production and elastase inhibitory activities of 80% methanol extract of Phellodendri cortex and its ethyl acetate fraction. Methods and Results: We prepared 80% methanol extract, and its fractions from phellodendri cortex. The treatment of RAW 264.7 cell with 25㎍/㎖ 80% methanol extract and ethyl acetate fraction resulted in no toxicity. We conducted assays of nitric oxide (NO) production and elastase inhibition. In the NO production assay, the ethyl acetate fraction showed an inhibitory effect approximately 17 times stronger than the 80% methanol extract. In elastase inhibitory assay, the ethyl acetate fraction also showed a stronger effect than the 80% methanol extract. In order to standardize the extract and fraction, we used TLC to separate the extract and observed the plate under UV light. We confirmed that the known pharmacological ingredients berberine, and palmatine in the 80% methanol extract and the ethyl acetate fraction. Conclusions: These results indicated that phellodendri cortex extract and its ethyl acetate fraction produced strong inhibitory effect on elastase and NO production.

Background : Ganoderma lucidum is a non-toxic, medicinal mushroom, which is known to possess anti-inflammatory and immunomodulating activities. However, the effects and mechanism of action of Ganoderma lucidum on lipopolysaccharide (LPS) and interferon-gamma (IFN-γ)-induced nitric oxide (NO) production and its-related cytokine expression are not yet fully understood. This study aimed to evaluate the effect of Ganoderma lucidum on NO production and NO-mediated pro-inflammatory cytokine expression in LPS/IFN-γ-induced RAW 264.7 macrophage-like cells. Methods and Results : The results showed that Ganoderma lucidum inhibited inducible NO synthase (iNOS) expression of RAW 264.7 macrophage-like cells at non-cytotoxic concentrations probably through the reduction of reactive oxygen species (ROS) production. After pre-treatment of cells with non-toxic doses of Ganoderma lucidum; NO production was significantly decreased. Moreover, Ganoderma lucidum treatment suppressed LPS/IFN-γ -stimulated pro-inflammatory cytokine secretion, including interleukin-1β and interleukin-6, in a dose-dependent manner. Conclusion : Taken together, these results indicate that the anti-inflammatory activation of Ganoderma lucidum in LPS/IFN-γ-stimulated macrophages might be due to abrogation of NO-dependent cytokine release by impairment of iNOS expression via ROS generation.

Background: In this study, we investigated the antibacterial and nitric oxide (NO) production inhibitory activities of 75% ethanol extract of Prunus sargentii branches and its fractions against acne pathogens. Methods and Results: The antibacterial activity against acne causing pathogens was determined using the disc diffusion assay. The ethyl acetate fraction showed higher activities against Propionibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis than those shown by other fractions. In the DPPH radical and NO scavenging assays, the butanol fraction showed strong DPPH radical and NO scavenging abilities. These activities were related to the total polyphenol and flavonoid contents of butanol fraction. On the other hand, the chloroform and ethyl acetate fractions exhibited the highest NO production inhibitory activity in Lipopolysaccharide (LPS)-stimulated Raw 264.7 cells compared to those exhibited by other fractions. Conclusions: The extract and its ethyl acetate fraction from the branches of P. sargentii exhibited antibacterial activity and could be used as functional materials in antimicrobial related fields. Moreover, the chloroform and ethyl acetate fractions are potential antiinflammatory agents and butanol fraction acts as an effective radical scavenger.

Arabis glabra is a localized common rhizomatous flowering plant, This plant is often used in Korean traditional systems of medicine as a remedy for blood cleaning, detoxification, abscess, gastrospasm, arthritis, contraction and diarrhea. Generally drugs that are used for arthritis have antinociceptive and anti-inflammatory properties. However, validity of the anti-inflammatory activity has not been scientifically investigated so far. Therefore, the aim of this study was to investigate the anti-inflammatory potential of A. glabra using the ethanolic extract and its sub-fractions. To evaluate the anti-inflammatory effects, we examined the inflammatory mediators such as nitric oxide (NO) and prostaglandin E2 (PGE2) on RAW 264.7macrophages. Our results indicated that hexane and chloroform fraction significantly inhibited the LPSinduced NO and PGE2 production in the cells. The hexane fraction inhibitory activity for NO tests with IC50 values showed in 21.0 ㎍/㎖. The chloroform fraction inhibitory activity for PGE2 tests with IC50 values showed in 18.0 ㎍/㎖. These efficacy are expected to be able to present the potential for the development of health functional food for the prevention inflammatory diseases because it has sufficient preventive medical possibilities. Further, it is determined that it is necessary to further study the mechanism of cytokine and protein expression associated with inflammation.