Humulus japonicus (HJ) is a widely used herbal medicine for pulmonary tuberculosis, hypertension, leprosy, and venomous wounds in Asia, particularly in China. Although HJ has certain physiological activities, such as longitudinal bone growth, antioxidation and alleviation of rheumatism, its anticancer activities, other than in colorectal and ovarian cancer, are yet to be studied. In this study, we investigated the anti-cancer activity and mechanism of methanol extracts of HJ (MeHJ) against human FaDu hypopharyngeal squamous carcinoma cells. MeHJ suppressed FaDu cell viability without affecting normal cells (L929), which was demonstrated using the MTT and Live & Dead assays. Furthermore, MeHJ effectively inhibited colony formation of FaDu cells, even at non-cytotoxic concentrations, and significantly induced apoptosis through the proteolytic cleavage of caspase-9, -3, -7, poly (ADP-ribose) polymerase and through the downregulation of BCL-2 and upregulation of BAX in FaDu cells, as determined by DAPI staining, flow cytometry, and western blot analyses. Collectively, these findings suggest that the inhibitory effects of MeHJ on the growth and colony formation of oral cancer cells may be mediated by caspase- and mitochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, MeHJ has the potential to be used as a natural chemotherapeutic drug against human oral cancer.
Asarum sieboldii Miq. (Aristolochiaceae) is a perennial herbaceous plant and has been used as traditional medicine for treating diseases, cold, fever, phlegm, allergies, chronic gastritis, and acute toothaches. Also, it has various biological activities, such as antiallergic, antiinflammatory, antinociceptive, and antifungal. However, the anticancer effect of A. sieboldii have been rarely reported, except anticancer effect on lung cancer cell (A549) of water extracts of A. sieboldii . This study investigated the anticancer activity of methanol extracts of A. sieboldii (MeAS) and the underlying mechanism in human FaDu hypopharyngeal squamous carcinoma cells. MeAS inhibited FaDu cells grown dose-dependently without affecting normal cells (L929), as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyl tetrazolium bromide and live and dead assay. In addition, concentration of MeAS without cytotoxicity (0.05 and 0.1 mg/mL) inhibited migration and colony formation. Moreover, MeAS treatment significantly induced apoptosis through the proteolytic cleavage of caspase-3, -7, -9, poly (ADP-ribose) polymerase, and downregulation of Bcl-2 and upregulation of Bax in FaDu cells, as determined by fluorescence-activated cell sorting analysis, 4`6-diamidino- 2-phenylindole stain, and western blotting. Altogether, these results suggest that MeAS exhibits strong anticancer effects by suppressing the growth of oral cancer cells and the migration and colony formation via caspase- and mitochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, MeAS can serve as a natural chemotherapeutic for human oral cancer.
Ficus carica L. (fig ) is one of the first cultivated crops and is as old as humans. This plant has been extensively used as a traditional medicine for treating diseases, such as cough, indigestion, nutritional anemia, and tuberculosis. However, the physiological activity of fig leaves on oral cancer is as yet unknown. In this study, we investigated the anticancer effect of methanol extracts of Ficus carica (MeFC) and the mechanism of cell death in human FaDu hypopharyngeal squamous carcinoma cells. MeFC decreased the viability of oral cancer (FaDu) cells but did not affect the viability of normal (L929) cells, as determined by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay and Live and Dead assay. In addition, MeFC induced apoptosis through the proteolytic cleavage of procaspase-3, -9, poly (ADP-ribose) polymerase (PARP), downregulation of Bcl-2, and upregulation of Bax, as determined by 4′,6-diamidino-2-phenylindole dihydrochloride staining and western blot analysis. Moreover, a concentration of MeFC without cytotoxicity (0.25 mg/mL) significantly suppressed colony formation, a hallmark of cancer development, and completely inhibited the colony formation at 1 mg/mL. Collectively, these results suggest that MeFC exhibits a potent anticancer effect by suppressing the growth of oral cancer cells and colony formation via caspase- and mitochondrial-dependent apoptotic pathways in FaDu human hypopharyngeal squamous carcinoma cells. Therefore, the methanol extract of Ficus carcica leaves provide a natural chemotherapeutic drug for human oral cancer.
In this study, we investigated the protective effects of Ulva lactuca methanol extracts against ultraviolet B (UVB)-induced DNA damage in HaCaT cells. First, the contents of general and antioxidative nutrient contents of Ulva lactuca were measured. The moisture, carbohydrate, crude protein, crude fat and ash were 14.01%, 44.80%, 23.19%, 3.10% and 14.90%, respectively. Magnesium that acts as DNA repair enzyme cofactor was the most abundant mineral followed by Ca, P and Fe. The total phenolic and anthocyanoside contents of Ulva lactuca were 2.69 mg/g and 0.13 mg/g, respectively. Cells treated with Ulva lactuca methanol extracts for 24 hours post UVB exposure increased cell viability in a concentration-dependent manner compared to the non-treated control. Also, Ulva lactuca methanol extracts decreased the levels of UVB-induced DNA damage such as cyclobutane pyrimidine dimer and DNA damage response (DDR) proteins such as p-p53 and p21. These results suggest that Ulva lactuca methanol extracts comprising physiological active substances such as Mg, polyphenols and anthocyanosides promote DNA repair by regulating genes related with DDR.
Codium fragile (Suringar) Hariot is an edible green seaweed that belong to the Codiaceae family and has been used in Oriental medicine for the treatment of enterobiasis, dropsy, and dysuria. Methanol extract of codium fragile has anti-oxidant, anti-inflammatory and anti-cancer properties, although the anti-cancer effect on oral cancer has not yet been reported. In this study, we investigated the anti-cancer activity and the mechanism of cell death by methanol extracts of Codium fragile (MeCF) on human FaDu hypopharyngeal squamous carcinoma cells. Our data showed that MeCF inhibits cell viability in a dose-dependent manner, and markedly induced apoptosis, as determined by the MTT assay, Live/Dead assay, and DAPI stain. In addition, MeCF induced the proteolytic cleavage of procaspase -3, -7, -9 and poly(ADP-ribose) polymerase(PARP), and upregulated or downregulated the expression of mitochondrial-apoptosis factor, Bax(pro-apoptotic factor), and Bcl-2(anti-apoptotic factor), . Futhermore, MeCF induced a cell cycle arrest at the G1/S phase through suppressing the expression of the cell cycle cascade proteins, p21, CDK4, CyclinD1, and phospho-Rb. Taken together, these results indicated that MeCF inhibits cell growth, and this inhibition is mediated by caspase- and mitochondrial-dependent apoptotic pathways through cell cycle arrest at the G1/S phase in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, methanol extracts of Codium fragile can be provided as a novel chemotherapeutic drug due to its growth inhibition effects and induction of apoptosis in human oral cancer cells.
치자 씨의 에틸 아세테이트, 증류수 및 70% 메탄올(v/v)의 3가지 용매를 사용한 추출물의 용매 별 플라보노이드 함량 및 항산화 활성, 금속 킬레이트 능력 측정을 통하여 치자의 기능성 식품 재료로서의 가치를 검토하기 위하여 본 실험을 수행하였다. 치자 씨의 탄닌 함량은 1.517±0.003 mg CE/g (mg of catechin equivalents) 건조 중량으로 나타났으며, 치자 씨 용매 별 추출 수율은 증류수 (36.61%), 70% 메탄올(30.10%), 에틸아세테이트(20.40%) 순으로 나타났다. 추출 용매 별 항산화 활성은 농도(0.2, 0.4, 0.6 mg/mL)가 증가할수록 유의적으로 증가하였으며(p<0.05), 양성대조구로 사용된 ascorbic acid, BHA, EDTA 보다 낮은 활성이 관찰되었다. 플라보노이드 함량(mg QE/g, mg of quercetin equivalents)은 70% 메탄올(0.799), 증류수(0.565), 에틸아세테이트(0.117) 순으로 관찰되었으며, 항산화 활성은 대부분의 분석에서 용매 별 플라보노이드 함량 순과 비슷한 양상으로 나타났지만 ABTS 라디칼 소거능과 SOD 유사활성은 증류수 추출물이 70% 메탄올 추출물의 항산화 활성 보다 높은 것으로 나타났다. 치자 씨의 각 용매별 수율과 항산화 활성을 비교하였을 때, 증류수와 70% 메탄올 용매를 사용하여 추출 이용하는 것이 바람직하다고 생각된다. 본 실험 결과 치자 씨는 플라보노이드 화합물을 다량 함유하고 있으며 그로 인한 높은 수준의 항산화 활성과 생리활성을 가지고 있어 기능성 식품 및 천연 항산화제로서의 가치가 매우 기대된다.
The effects of antioxidant measured by DPPH radical scavenger activity, the extract of harvestry in July shown highest activity(91.8±0.01%). The all extracts of 4 parts(leaf, f1ower, root, branch) of harvestry in July were shown better radical scavenger activity than BHT(89.2±0.01%). The antibacteria activity of Agrimonia pilosa extracts was determined by using a paper disc method against bacteria, the methanol extracts(70%) showed the most active antibacteria activity against 12 kinds of harmful microorganisms. The methanol extracts(70%) showed the highest antibacterial activity against S. aureus and B. subtilis. In addition, the minimum inhibitory concentration(MIC) of methanol extracts(70%) against S. aureus, B. subtilis, C. diphtheriae, S. mutans, and S. epidermidis were revealed 0.625~10mg/mL. As a result, antibacteria activity in methanol(70%) extract of Agrimonia pilosa shown higher gram positive bacteria than gram negative bacteria.
This study aimed to evaluate the efficacy of the antioxidative and antidiabetic activities of the flowers, leaves, and roots of the Jerusalem artichoke (Helianthus tuberosus L.). The total polyphenol and flavonoid contents of the leaves were higher than those of the flowers and roots. However, the DPPH radical-scavenging and hydroxyl radical-scavenging activities of the flowers were higher than those of the leaves and roots. The nitrite-scavenging ability under acidic conditions was high in Jerusalem artichoke flower extracts. The α-glucosidase inhibitory activity and α-amylase inhibitory activity of a methanol extract of Jerusalem artichoke roots were about 60% (5 mg/mL concentration). Based on these experiments, it can be concluded that the flowers leaves, and roots of the Jerusalem artichoke can be used as natural preservatives. Therefore, they can be developed as functional foods, to take advantage of their antioxidant activity and abundant polyphenols. This study suggests that the whole Jerusalem artichoke, including roots, leaves, and flowers, is useful as a functional, nutritious food product.
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.
본 연구에서는 기능성 식품 및 화장품 소재로써 갈색 느티만가닥버섯(Hypsizygus marmoreus)의 이용가능성을 조사하기 위해서 갈색 느티만가닥버섯 메탄올 추출물의 항산화 활성 및 tyrosinase 저해 효과를 갓(pileus)과 대(stipes)로 분리하여 부위별로 조사하였다. 메탄올 추출물 중 갓과 대의 총 폴리페놀 함량은 각각 8.7±3.27ug/mg과 5.6±2.85ug/mg이었고플라보노이드 함량은 각각 2.8±3.81ug/mg과 1.4±1.95ug/mg이었으며 총 폴리페놀과 플라보노이드 함량 모두 대보다 갓에서 높게 나타났다. Tyrosinase 저해 활성은 추출물의 농도에 따라 증가하는 경향을 나타내었으나 양성 대조구로 사용한 2% 알부틴(arbutin)과 비교했을 때 1200mg/ml의 고농도에서도 갓은66.9%, 대는 57.97%의 낮은 저해 활성을 나타내었다.항산화 활성은 DPPH에 의한 라디칼소거 활성을 측정하여 확인하였으며 갓과 대의 DPPH 라디칼 소거능은 20mg/ml의 농도에서도 각각 52.55%와 30.35%로 낮게 나타났다. 추출물이 B16BL6 mouse melanomacell에 미치는 영향을 알아보기 위해 WST-1 assay (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulphonate)를 이용하여 세포생존율을 조사하였으며 갓과 대 모두 200-2,000ug/ml의 농도로처리하였을 때 100% 이상의 생존율을 나타내었고추출물의 처리 농도가 증가함에 따라 세포생존율도증가하는 경향을 나타내었으며 양성대조군인 0.04%adenosine을 처리한 경우보다 높게 나타났다.
Oxidative stress has been reported to be one of causes of neuritis. This study examined antioxidative activities of methanol extracts of six amphibian species known to be medicinal animals (Rana catesbeiana, R. coreana, R. rugosa, R. dybowskii, R. nigromaculata, and Hyla japonica) and investigated their effects of inhibiting nitric oxide (NO) production and cytotoxicity on the murine macrophage RAW264.7 cells. As inflammation is closely associated with reactive oxygen species, assays on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, xanthine oxidase inhibitory activity, superoxide anion radical scavenging activity and NO scavenging activity of the extracts of the six species were performed to investigate their antioxidative activity. The results obtained were as follows; All extracts showed antioxidative activity, and the activity of R. dybowskii was the highest in comparison among those. Anti-inflammatory effects of the extracts were also examined, the five extracts except that of R. rugosa did not show cytotoxicity for RAW264.7 cells at the maximal concentration (1,000 μg mL-1). Selectivity index, meaning NO scavenging activity compared to cytotoxicity, showed the highest level in the extract of R. dybowskii. These results will be very useful basic data for future studies on prevention and treatment of human diseases to understand the biological roles of amphibian extracts throughout the antioxidative or anti-inflammatory pathways.
This study was conducted to investigate the physiological activities of the water and ethanol extracts from three strains of Hypsizygus marmoreus. Largely, anti-hypertensive, anti-inflammatory, anti-diabetic, anti-tumorigenic activities were carried out. In addition, β-glucan contents were also measured. Mangadak 2 ho has the 36%(w/w) β-glucan contents, it is highest value of all strains. And it also showed highest inhibitory activities of angiotensin converting enzyme. GONJI No.6 highly inhibited releasing of nitric oxide from Raw 264.7. Anti-diabetic activity, the Hypsizygus marmoreus(from Japan) showed highest inhibitory activity on amyloglucosidase. MTT assay was performed on the anti-tumorigenic test. The cancer cell were used to HeLa, A549. repectively.
This study was conducted to investigate the physiological activities of the water and methanol extracts from Auricularia auricula-judae(black and brown) and Auricularia polytricha. Largely, anti-oxidant, anti-tumorigenic, anti-inflammatory, anti-diabetic activities were carried out. DPPH scavenging activity was performed on the anti-oxidant test. MTT assay and nitrite scavenging acitivity were performed on the anti-tumorigenic test. The cancer cell were used to HeLa, A549, HT-29 and MCF-7 and normal cell was used as control. lastly, in the anti-inflammatory and anti-diabetic activities, NO assay and α-amyloglucosidase inhibition were performed, repectively.
Antioxidant activities of Hericium erinaceum was in vestigated in subfractions of methanol extract such as hexane, chloroform, ethylacetate, butanol and water subfractions. Chlorofrom fraction exhibited the highest antioxidant activity and showed the highest level of total phenolic contents. The level of total phenolics may be correlated with the antioxidant activities of the subfractions. Therefore, the phenolic compound in the chloroform subfractions exhibiting high antioxidant acticity was identified by mass spectrometry using LC-MS/MS. In addition, hot water extract of this mushroom were tested for antimutagenic activity by Ames Salmonellaassay.Thisextractshowed inhibitiory effect on the mutagenesis induced in Salmonella typhimurium strain TA100 and TA98 by the directly actingmutagen 4-nitroquinoline-1-oxide(0.15μg/plate) and sodium azide (0.5μg/plate). The extract also inhibited mutagenesis induced in Salmonella typhimurium strain TA100 and TA98 by the indirectly actingmutagen 2-aminoanthracen(2μg/plate) at 5000μg/plate and 200 μg/plate, respectively.
To find an alternative for synthetic pesticides, methanol extract from plant samples were tested for their insecticidal activity against insect. The extract of Asiasarum sieboldii had strongly insecticidal activity against Plutella xylostella. Roots of A. sieboldii were extracted with methanol, and the concentrated extract was partitioned with n-hexane, ethylacetate, n-buthanol and H2O. The highest activity was shown in the hexane fraction. Activity-guided fractionation led to the isolation of two amides from hexane fration through the repeated silica gel column chromatographic separations. From the interpretation of spectropic data including NMR, MS, IR, the chemical structures of compounds were determined as dodeca-2E,4E,8Z,10Z-tetraenoic acid isobutylamide and dodeca-2E,4E,8Z, 10E-tetraenoic acid isobutylamide. These compounds showed insecticidal activity on P. xylostella by 96.7% at 100ppm. The liquid formulation controlled on cabbage effectively. The extract and compounds from A. sieboldii showed insecticidal activity against Nilaparvata lugens. As a naturally occurring pesticide, A. sieboldii could be useful as a new botanic insecticide.
곰취(Ligularia fischeri)의 이용성을 높이데 필요한 기초자료를 확보하고자 메탄올을 용매로 하여 추출 온도 및 시간에 따른 생리활성 효과를 조사하였다. 곰취 메탄올추출물 1,000mg·L-1에 함유된 총 페놀함량, 총플라보노이드 함량은 각각 75.8-297.7mg·L-1 and 45.6-173.6mg·L-1이었다. 추출온도와 시간은 총 페놀함량, 총 플라보노이드 함량, 전자공여 능 의 경우 95℃에서 6시간 동안 추출했을 때 가장 좋았다. 그러나 아질산염 소거능은 75℃에서 12시간 추출한 것에서 97.4%로 가장 높게 나타났다. 곰취 메탄을 추출물 200mg·L-1 및 400mg·L-1 농도는 각각 폐암과 위암세포 증식을 90% 이상 억제시켰다. 따라서 곰취는 높은 생리활성 기능을 나타내는 채소로 나타났으며, 곰취를 가공품으로 이용하기 위해 메탄올로 추출할 때는 95℃에서 6시간 정도 하는 것이 좋을 것으로 생각되었다.
The effects of the Rhynchosia nulubilis extracts on serum lipid concentrations were evaluated in rats. Forty-two male Sprague-Dawley rats weighing 100±10g were divided into six groups and fed high fat and high cholesterol diets for four weeks. Experimental groups were administered with following diets; basal fat diet(control) and basal and high fat diets with 1% cholesterol and Rhynchosia nulubilis extract(40mg/kg, 400mg/kg and 4000mg/kg). The concentrations of serum triglyceride in rats fed the Rhynchosis mulubilis extract, high fat, 1% cholesterol, and RN-HFC(-40, -400, -4000mg/kg) groups were lower than those in other groups. The concentrations of total cholesterol in the RN-HFC (-40, -400, -4000mg/kg) groups were lower than those in fat diet groups. The concentrations of HDL-cholesterol in serum of the RN-HFC(-40, -400, 4000mg/kg) groups were significantly higer than those of other groups. The levels of LDL-cholesterol in serum of the RN-HFC(-40, -400, -4000mg/kg) groups were tended to be lower than those of other groups. These results suggest that Rhynchosia nulubilis extract may reduce elevated levels of serum lipid concentrations in rats fed high fat diets.