• Title/Summary/Keyword: In vitro assays

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Water-extract of Helianthus annuus Seed Exhibits Potent Anti-asthma Activity In Vitro and In Vivo (해바라기씨 추출물의 천식 완화 효과)

  • Heo Jin-Chul;Park Ja-Young;Woo Sang-Uk;Chung Shin-Kyo;Jeong Kyu-Shick;Lee Jin-Man;Ma Jin-Yeol;Lee Sang-Han
    • Food Science and Preservation
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    • v.13 no.4
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    • pp.495-500
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    • 2006
  • In order to investigate whether antioxidant biomaterials inhibits IL-4 and/or IL-13 expression in vitro and in vivo, we carried out antioxidant assays by enzyme or cell-based assays with Helianthus annuus extract. Antioxidant assays include DPPH, FRAP, hydroxyl radical assays. Helianthus annuus extract exhibited SOD scavenging activity, and had different patterns by each solvent extracted reaction. DW extract inhibited oxidative stress by $H_2O_2$ that induced apoptosis. We measured $CD4^+$ cell and IL-/13 cytokine expression in a classical mouse animal model. The result show that Helianthus annuus extract showed strung inhibition of immune response in the lung. These result suggest that Helianthus annuus extract can reduce inflammation induced by n mouse asthma model.

In Vitro and In Vivo Antioxidant Activity of Aged Ginseng (Panax ginseng)

  • Chung, Soo Im;Kang, Mi Young;Lee, Sang Chul
    • Preventive Nutrition and Food Science
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    • v.21 no.1
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    • pp.24-30
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    • 2016
  • Fresh ginseng roots were aged in an oven at $80^{\circ}C$ for 14 d. The in vitro and in vivo antioxidant activities of this aged ginseng, in comparison with those of the white and red ginsengs, were evaluated. In in vitro antioxidant assays, the ethanolic extracts from aged ginseng showed significantly higher free radical scavenging activity and reducing power than those of the white and red ginsengs. In in vivo antioxidant assays, mice were fed a high fat diet supplemented with white, red, or aged ginseng powders. High fat feeding resulted in a significant increase in lipid peroxidation and a substantial decrease in antioxidant enzymes activities in the animals. However, diet supplementation of ginseng powders, particularly aged ginseng, markedly reduced lipid peroxidation and enhanced the antioxidant enzymes activities. The results illustrate that the aged ginseng has greater in vitro and in vivo antioxidant capacity than the white and red ginsengs. The aged ginseng also showed considerably higher total saponin, phenolic, and flavonoid contents, indicating that its antioxidant capacity may have been partly due to its high levels of antioxidant compounds. This new ginseng product may be useful as a functional food with strong antioxidant potential.

Oxidative stability of extracts from red ginseng and puffed red ginseng in bulk oil or oil-in-water emulsion matrix

  • Lee, Sang-Jun;Oh, Sumi;Kim, Mi-Ja;Sim, Gun-Sub;Moon, Tae Wha;Lee, JaeHwan
    • Journal of Ginseng Research
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    • v.42 no.3
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    • pp.320-326
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    • 2018
  • Background: Explosive puffing can induce changes in the chemical, nutritional, and sensory quality of red ginseng. The antioxidant properties of ethanolic extracts of red ginseng and puffed red ginseng were determined in bulk oil and oil-in-water (O/W) emulsions. Methods: Bulk oils were heated at $60^{\circ}C$ and $100^{\circ}C$ and O/W emulsions were treated under riboflavin photosensitization. In vitro antioxidant assays, including 2,2-diphenyl-1-picrylhudrazyl, 2,2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid, ferric reducing antioxidant power, total phenolic content, and total flavonoid content, were also performed. Results: The total ginsenoside contents of ethanolic extract from red ginseng and puffed red ginseng were 42.33 mg/g and 49.22 mg/g, respectively. All results from above in vitro antioxidant assays revealed that extracts of puffed red ginseng had significantly higher antioxidant capacities than those of red ginseng (p < 0.05). Generally, extracts of puffed red and red ginseng had high antioxidant properties in riboflavin photosensitized O/W emulsions. However, in bulk oil systems, extracts of puffed red and red ginseng inhibited or accelerated rates of lipid oxidation, depending on treatment temperature and the type of assay used. Conclusion: Although ethanolic extracts of puffed red ginseng showed stronger antioxidant capacities than those of red ginseng when in vitro assays were used, more pro-oxidant properties were observed in bulk oils and O/W emulsions.

Pre-clinical QT Risk Assessment in Pharmaceutical Companies - Issues of Current QT Risk Assessment -

  • Takasuna, Kiyoshi; Katsuyoshi, Chiba;Manabe, Sunao
    • Biomolecules & Therapeutics
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    • v.17 no.1
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    • pp.1-11
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    • 2009
  • Since the Committee for Proprietary Medicinal Products (CPMP) of the European Union issued in 1997 a "points to consider" document for the assessment of the potential for QT interval prolongation by non-cardiovascular agents to predict drug-induced torsades de pointes (TdP), the QT liability has become the critical safety issue in the development of pharmaceuticals. As TdP is usually linked to delayed cardiac repolarization, international guideline (ICH S7B) has advocated the standard repolarization assays such as in vitro IKr (hERG current) and in vivo QT interval, or in vitro APD (as a follow up) as the best biomarkers for predicting the TdP risk. However, the recent increasing evidence suggests that the currently used above biomarkers and/or assays are not fully predictive for TdP, but also does not address potential new druginduced TdP due to the selective disruption of hERG protein trafficking to the cell membrane or VT and/or VF with QT shortening. There is, therefore, an urgent need for other surrogate markers or assays that can predict the proarrhythmic potential of drug candidate. In this review, we provide an ideal pre-clinical strategy to predict the potentials of QT liability and lethal arrhythmia of the drug candidates with recent issues in this field in mind, not at the expense of discarding therapeutically innovative drugs.

In vitro antioxidant property and α-glucosidase and pancreatic lipase inhibiting activities of Jeju camellia mistletoe (Korthalsella japonica (Thunb.) Engl.) extracts (제주 동백나무 겨우살이(Korthalsella japonica (Thunb.) Engl.)의 항산화 및 α-glucosidase와 pancreatic lipase 저해 활성)

  • Park, Eun Mi;Kim, Min Young
    • Journal of Applied Biological Chemistry
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    • v.60 no.3
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    • pp.241-244
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    • 2017
  • The antioxidant activity of various solvent extracts from Jeju camellia mistletoe (Korthalsella japonica (Thunb.) Engl.) was investigated using various in vitro assays as the 1,1-diphenyl-2-picrylhydrazyl radical scavenging, ferrous ion chelating and reducing power assays. Methanol and ethanol extracts showed the most potent antioxidant activity in all assays tested followed by water extract. The inhibitory effect of the Jeju camellia mistletoe extracts on pancreatic lipase and $\acute{a}$-glucosidase was also evaluated and the results showed that methanol and ethanol extracts markedly reduced both enzyme activities. Therefore, the methanol and ethanol extracts of Jeju camellia mistletoe is definitely worthy of further investigation for these beneficial effects on nutraceutical medicine.

Isolation and Biological Activities of an Alkaloid Compound (3-methylcanthin-5, 6-dione) from Picrasma quassiodes (D. Don) Benn.

  • Yin, Yu;Lee, Seok-Ki;Wang, Myeong-Hyeon
    • Natural Product Sciences
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    • v.17 no.1
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    • pp.5-9
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    • 2011
  • An alkaloid, 3-methylcanthin-5, 6-dione, was isolated from the stem of Picrasma quassioides (D. Don) Benn. and characterized by comprehensive analyses of its 1D and 2D NMR spectra. It was also evaluated for its cytotoxic activity in vitro against three human cancer cell lines (MDA-MB-231, HT-29 and NCI-N87), using MTT assays. We found that 3-methylcanthin-5, 6-dione exhibited significant anti-inflammatory activity via inhibiting NO production induced in LPS-stimulated murine macrophage RAW264.7 cells. The antioxidant activity of 3-methylcanthin-5, 6-dione was measured by DPPH free radical scavenging assays, hydroxyl radical scavenging assays and reducing power assays. Our results showed that 3-methylcanthin-5, 6-dione has significant biological activities.

Chloroquine and Valproic Acid Combined Treatment in Vitro has Enhanced Cytotoxicity in an Osteosarcoma Cell Line

  • Wang, Chuan-Kun;Yu, Xi-Dong;Li, Qiang;Xie, Gang;Teng, Yue
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.8
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    • pp.4651-4654
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    • 2013
  • Choroquine (CQ) and valproic acid (VPA) have been extensively studied for biological effects. Here, we focused on efficacy of combined CQ and VPA on osteosarcoma cell lines. Viability of osteosarcoma cell lines (U20S and HOS) was analyzed by MTT assay. Apoptotic assays and colony formation assays were also applied. ROS generation and Western Blotting were performed to determine the mechanism of CQ and VPA combination in the process of apoptosis. The viability of different osteosarcoma cell lines significantly decreased after CQ and VPA combination treatment compared with either drug used alone, and apoptosis was increased significantly. ROS generation was triggered leading to expression of apoptosis related genes being increased and of antiapoptotic related genes being decreased. From our data shown here, CQ and VPA combination treatment in vitro enhanced cytotoxicy to osteosarcoma cells.

In Vitro Studies on the Genotoxic Effects of Wood Smoke Flavors

  • Chung, Young-Shin;Ahn, Jun-Ho; Eum, Ki-Hwan;Choi, Seon-A;Oh, Se-Wook;Kim, Yun-Ji;Park, Sue-Nie;Yum, Young-Na;Kim, Joo-Hwan;Lee, Michael
    • Toxicological Research
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    • v.24 no.4
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    • pp.321-328
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    • 2008
  • Smoke flavors based on the thermal decomposition of wood have been applied to a variety of food products as an alternative for traditional smoking. Despite its increasing use, the available genotoxicity data on wood smoke flavors (WSF) are still controversial. Thus, potential genotoxic effects of WSF in four short-term in vitro genotoxicity assays were investigated, which included the Ames assay, chromosomal aberration assay, micronucleus test and the alkaline comet assay. WSF did not cause any mutation in the Ames assay using five tester strains at six concentrations of 0.16, 0.31, 0.63, 1.25, 2.5 and 5 ${\mu}l/plate$. To assess clastogenic effect, the in vitro chromosomal aberration assay was performed using Chinese hamster lung cells. No statistically significant increase in the number of metaphases with structural aberrations was observed at the concentrations of 1.25, 2.5, and 5 ${\mu}l/ml$. The in vitro comet assay and micronucleus test results obtained on L5178Y cells also revealed that WSF has no genotoxicity potential, although there was a marginal increase in micronuclei frequencies and DNA damage in the respective micronucleus and comet assays. Taken together, based on the results obtained from these four in vitro studies, it is concluded that WSF is not a mutagenic agent in bacterial cells and causes no chromosomal and DNA damage in mammalian cells in vitro.

Comparative evaluation of the mutagenicity and genotoxicity of smoke condensate derived from Korean cigarettes

  • Kim, Ha Ryong;Lee, Jeong Eun;Jeong, Mi Ho;Choi, Seong Jin;Lee, Kyuhong;Chung, Kyu Hyuck
    • Environmental Analysis Health and Toxicology
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    • v.30
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    • pp.14.1-14.7
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    • 2015
  • Objectives Cigarette smoking is associated with carcinogenesis owing to the mutagenic and genotoxic effects of cigarette smoke. The aim of this study was to evaluate the mutagenic and genotoxic effects of Korean cigarettes using in vitro assays. Methods We selected 2 types of cigarettes (TL and TW) as benchmark Korean cigarettes for this study, because they represent the greatest level of nicotine and tar contents among Korean cigarettes. Mutagenic potency was expressed as the number of revertants per ${\mu}g$ of cigarette smoke condensate (CSC) total particulate matter whereas genotoxic potency was expressed as a concentration-dependent induction factor. The CSC was prepared by the International Organization for Standardization 3308 smoking method. CHO-K1 cells were used in vitro micronucleus (MNvit) and comet assays. Two strains of Salmonella typhimurium (Salmonella enterica subsp. enterica ; TA98 and TA1537) were employed in Ames tests. Results All CSCs showed mutagenicity in the TA98 and TA1537 strains. In addition, DNA damage and micronuclei formation were observed in the comet and MNvit assays owing to CSC exposure. The CSC from the 3R4F Kentucky reference (3R4F) cigarette produced the most severe mutagenic and genotoxic potencies, followed by the CSC from the TL cigarette, whereas the CSC from the TW cigarette produced the least severe mutagenic and genotoxic potencies. Conclusions The results of this study suggest that the mutagenic and genotoxic potencies of the TL and TW cigarettes were weaker than those of the 3R4F cigarette. Further study on standardized concepts of toxic equivalents for cigarettes needs to be conducted for more extensive use of in vitro tests.

Anti-oxidant and Anti-tumor Activities of Crude Extracts by Gastrodia elata Blume (천마추출물의 항산화 및 항암 활성)

  • Heo Jin-Chul;Park Ja-Young;An Sang-Mi;Lee Jin-Man;Yun Chi-Young;Shin Heung-Mook;Kwon Taeg-Kyu;Lee Sang-Han
    • Food Science and Preservation
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    • v.13 no.1
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    • pp.83-87
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    • 2006
  • Gastrodia elata Blume is a major imp0l1ant medicinal resource in Korea. In order to confirm the biological activities of Gastrodia elata Blume, we carried out various in vitro assays. Of them, anti-oxidant and anti-tumor activities were detected from assays. The prototype of Gastrodia elata Blume extracts was used for 1he evaluation of DPPH, FRAP, hydroxyradical scavenging assay as anti-oxidant assays, as well as anti-tumor asctivities as wound assay and invasion assay. As a result, the prototype of Gastrodia elata Blume extracts showed potent anti-oxidative activity and anti-tumor activity in vitro. These above results suggest that 1he Gastrodia elata Blume extracts could have potential to alleviate oxidation process, cell motility activity, and tumorigenesis.