• Title/Summary/Keyword: Biological Assay

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Enhancement of Anti-inflammatory Activity by Fermentation of Sargassum siliquanstrum (꽈배기모자반의 발효를 통한 항염증 활성의 증진)

  • Lee, Sol-Ji;Lee, Dong-Geun;Kim, Mihyang;Kong, Chang-Suk;Yu, Ki-Hwan;Kim, Yuck-Young;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.26 no.3
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    • pp.318-324
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    • 2016
  • This study was aimed to verify anti-inflammatory activity of fermented Sargassum siliquanstrum with lactic acid bacteria. Anti-inflammatory activities were compared by measuring the amount of nitric oxide (NO) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and suppressive effect on inducible nitric oxide synthase (iNOS) expression in stably transfected RAW 264.7 cells. Inhibitory activities of NO production and iNOS expression were measured after confirmation of NO radical scavenging activities. Fermentation increased NO radical scavenging activities from 7.6% to 15.2% compared to non-fermented condition, and fermentation with Lactobacillus sp. SH-1 was the most efficient. Fermentation without algal debris showed better NO radical scavenging activities than that with debris. Fermentation with Lactobacillus sp. SH-1 also showed the highest NO production inhibitory activity (64.1%) in LPS-stimulated RAW 264.7 cells. LPS-induced iNOS expression was diminished to 28.6, 35.6, 49.4 and 58.5 at 50, 100, 500 and 1,000 μg/ml, respectively, by fermentation with Lactobacillus sp. SH-1. According to MTT assay, fermented S. siliquanstrum did not influence the cell viability at all concentrations tested, meaning no or less cytotoxicity. These results suggest that S. siliquanstrum has NO radical scavenging activity and anti-inflammatory activity. Thus biological activities of S. siliquanstrum were upgraded by fermentation, which could be used for the development of functional foods.

Biological Activities of Isolated Icariin from Epimedium koreanum Nakai (삼지구엽초로부터 분리한 Icariin의 생리활성)

  • Kim, Seo-Jin;Park, Myoung-Su;Ding, Tian;Wang, Jun;Oh, Deog-Hwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1397-1403
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    • 2011
  • Epimedium koreanum Nakai is a wild medicinal plant commonly consumed in South Korea due to its health beneficial effects. In the present study, the antioxidative, antimutagenic and immunological activities of E. koreanum Nakai extracts were investigated for their use in food. The yields of icariin compounds from the ethanol extract as well as the ethyl acetate, butanol, hexane, water, and chloroform fractions of E. koreanum were 27.9, 2.5, 1.7, 1.4, and 1.3 ${\mu}g/g$, respectively. The icariin components (295.5 ${\mu}g/g$) were collected from the ethyl acetate fraction by thin layer chromatography (TLC) and analyzed via high performance liquid chromatography (HPLC). The antioxidant activities of each fraction were as follows: ethyl acetate (49.0 ${\mu}g/mL$), butanol (59.2 ${\mu}g/mL$), hexane (119.8 ${\mu}g/mL$), water (122.0 ${\mu}g/mL$), and chloroform (138.5 ${\mu}g/mL$), based on $RC_{50}$ ${\mu}g/mL$. Icariin, isolated and identified as the main component, showed strong antioxidant activity with a $RC_{50}$ value of 15.3 ${\mu}g/mL$, which was higher than those of ascorbic acid (19.5 ${\mu}g/mL$) and ${\alpha}$-tocopherol (18.2 ${\mu}g/mL$). In an Ames test, none of the fractions produced mutagenic effects on Salmonella Typhimurium TA98 and TA100. In an immunomodulating activity test, the effects of E. koreanum Nakai on B cells (Rhamos) and T cells (Jurkat) were investigated. These results show that the growth and viability of B and T cells were increased by isolated icariin components for 1.27 and 1.28 fold, respectively. These results also provide preliminary data for the development of E. koreanum Nakai as an edible food material.

Mitigation Effects of Foliar-Applied Hydrogen Peroxide on Drought Stress in Sorghum bicolor (과산화수소 엽면 처리에 의한 수수에서 한발 스트레스 완화 효과)

  • Shim, Doo-Do;Lee, Seung-Ha;Chung, Jong-Il;Kim, Min Chul;Chung, Jung-Sung;Lee, Yeong-Hun;Jeon, Seung-Ho;Song, Gi-Eun;Shim, Sang-In
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.2
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    • pp.113-123
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    • 2020
  • Global climatic change and increasing climatic instability threaten crop productivity. Due to climatic change, drought stress is occurring more frequently in crop fields. In this study, we investigated the effect of treatment with hydrogen peroxide (H2O2) before leaf development on the growth and yield of sorghum for minimizing the damage of crops to drought. To assess the effect of H2O2 on the growth of sorghum plant, 10 mM H2O2 was used to treat sorghum leaves at the 3-leaf stage during growth in field conditions. Plant height, stem diameter, leaf length, and leaf width were increased by 7.6%, 9.6%, 8.3% and 11.5%, respectively. SPAD value, chlorophyll fluorescence (Fv/Fm), photosynthetic rate, stomatal conductance, and transpiration rate were increased by 3.0%, 4.9%, 26.0%, 23.4% and 12.7%, respectively. The amount of H2O2 in the leaf tissue of sorghum plant treated with 10 mM H2O2 was 0.7% of the applied amount after 1 hour. The level increased to approximately 1.0% after 6 hours. The highest antioxidant activity measured by the Oxygen Radical Absorbance Capacity assay was 847.3 µmol·g-1 at 6 hour after treatment. However, in the well-watered condition, the concentration of H2O2 in the plant treated by the foliar application of H2O2 was 227.8 µmol·g-1 higher than that of the untreated control. H2O2 treatment improved all the yield components and yield-related factors. Panicle length, plant dry weight, panicle weight, seed weight per plant, seed weight per unit area, and thousand seed weight were increased by 8.8%, 18.0%, 24.4%, 24.7%, 29.9% and 7.1%, respectively. Proteomic analysis showed that H2O2 treatment in sorghum increased the tolerance to drought stress and maintained growth and yield by ameliorating oxidative stress.

Protective effects of Aruncus dioicus var. kamtschaticus extract against hyperglycemic-induced neurotoxicity (포도당 처리로 유도된 뇌신경세포 독성에 대한 눈개승마 추출물의 보호효과)

  • Park, Su Bin;Lee, Uk;Kang, Jin Yong;Kim, Jong Min;Park, Seon Kyeong;Park, Sang Hyun;Choi, Sung-Gil;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.49 no.6
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    • pp.668-675
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    • 2017
  • To assess the physiological effects of Aruncus dioicus var. kamtschaticus extract on cytoxicity of a neuronal cell line, antioxidant activity, and neuroprotection against intensive glucose-induced oxidative stress were quantitated. Compared to the other fractions, the ethyl acetate fraction of Aruncus dioicus var. kamtschaticus (EFAD) showed the highest total phenolics and flavonoids. The 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assay and malondialdehyde inhibitory effect test confirmed the superior antioxidant activity of EFAD. Moreover, EFAD also decreased the intracellular ROS level and suppressed neuronal cell death against intensive glucose- or $H_2O_2$-induced oxidative stress. Additionally, assessment of ${\alpha}$-glucosidase and acetylcholinesterase inhibitory activities revealed that EFAD was an effective inhibitor of ${\alpha}$-glucosidase and acetylcholinesterase. Finally, high-performance liquid chromatography analysis identified caffeic acid as the main ingredient of EFAD. Overall, these results suggest that the EFAD is a good natural source of biological compounds that counteract hyperglycemic neuronal defects.

Cellular Antioxidant Activity and Whitening Effects of Dendropanax morbifera Leaf Extracts (황칠나무 잎 추출물의 세포 항산화 활성과 미백활성 측정)

  • Park, Su Ah;Park, Jun;Park, Chan Il;Jie, Young Jong;Hwang, Yun Chan;Kim, Yong Hyun;Jeon, So Ha;Lee, Hye Mi;Ha, Ji Hoon;Kim, Kyeong Jin;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.41 no.4
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    • pp.407-415
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    • 2013
  • In this study, we investigated the antioxidant activities on HaCaT and the whitening effects on B16F1 melanoma cells of Dendropanax morbifera leaf extract. In an antioxidative activity assay using HaCaT cells, the ethyl acetate ($50{\mu}g/ml$) and aglycone fractions ($25{\mu}g/ml$) of the D. morbifera leaf extract didn't exhibit any characteristics of cytotoxicity. When HaCaT cells were exposed to a single large dose ($800mJ/cm^2$) of UVB, the extracts protected the cells against UVB radiation. When HaCaT cells were treated with 10 mM $H_2O_2$ and $4{\mu}M$ rose bengal, the ethyl acetate ($6.25{\sim}50{\mu}g/ml$) and aglycone ($6.25{\sim}25{\mu}g/ml$) fractions protected the cells against oxidative damage in a concentration dependent manner. When the whitening effects of D. morbifera leaf extract were tested in melanoma B16/F1 cells treated with the a-melanocyte stimulating hormone (${\alpha}$-MSH), the extracts inhibited ${\alpha}$-MSH-stimulated intra/extracellular melanogenesis in a concentration dependent manner. The inhibitory effects of the ethyl acetate and aglycone fractions of D. morbifera leaf extract were 21% and 44% at $25{\mu}g/ml$, respectively. Both are more effective than arbutin (15% at $25{\mu}g/ml$) which is known as a whitening agent. These results indicate that fractions of the D. morbifera leaf can function as cell protectants and natural antioxidants in biological systems, particularly skins exposed to UV radiation by quenching and/or scavenging $^1O_2$ and other ROS, and protecting cells against ROS. In addition, fractions of the D. morbifera leaf can be applied to new whitening cosmetics because of their inhibitory effects on ${\alpha}$-MSH stimulated melanogenesis in B16F1 melanoma cells.

Antioxidative Properties of Extract/Fractions of Suaeda asparagoides and Salicornia herbacea Extracts(I) (함초(나문재 및 퉁퉁마디) 추출물의 항산화 작용에 관한 연구(I))

  • Park, Soo-Nam;Jeon, So-Mi;Kim, So-I;Ahn, Jeung-Youb
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.3
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    • pp.145-152
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    • 2007
  • In this study, the antioxidative effects of Sueada asparagoides and Salicornia herbacea extracts were investigated. The free radical(1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity($FSC_{50}$) of extract/fractions of Sueada asparagoides was in the order: 100 % ethanol extract(329.33 ${\mu}g/mL$) < 50 % ethanol extract(40.73) < ethylacetate fraction(13.87) < deglycosylated aglycone fraction (7.80). In case of Salicornia herbacea, the free radical scavenging activities of ethylacetate fraction and aglycone fraction were 13.87 and 7.80 ${\mu}g/mL$, respectively. Reactive oxygen species(ROS) scavenging activities($OSC_{50}$) of Sueada asparagoides and Solicornia herbacea extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activity of Sueada asparagoides extracts was 50 % ethanol extract($OSC_{50}$, $0.99{\mu}g/mL$) < ethylacetate fraction (0.05) < aglycone fraction (0.03). Aglycone fraction showed the most prominent scavenging activity. In case of Salicornia herbacea, the ROS scavenging activities of ethylacetate fraction and aglycone fraction were 0.10 and 0.20 ${\mu}g/mL$, respectively. The protective effects of extract/fractions of Sueada asparagoides and Salicornia herbacea on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethanol extract(100%) of Sueada asparagoides diminished photohemolysis in a concentration dependent manner($1{\sim}100{\mu}g/mL$). Particularly deglycosylated aglycone fraction exhibited the most prominent celluar protective effect($\tau_{50}$, 310 min at 50 ${\mu}g/mL$). In case of Salicornia herbacea, ethylacetate fraction exhibited more potent protective effect. These results indicate that extract/fractions of Sueada asparagoides can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS.

Anti-oxidative Activities of Castanea crenata Leaf Extract/Fractions and Application on Cosmetics (밤나무 잎 추출물의 항노화 활성 및 화장품에의 응용( I ))

  • Kim, Jin-Young;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.4
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    • pp.259-268
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    • 2008
  • In this study, the antioxidative effects, inhibitory effects on tyrosinase and elastase and components of Castanea crenata leaf were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl radical, DPPH) scavenging activity ($FSC_{50}$) of extract / fractions of Castanea crenata left was in the order: 50% ethanol extract ($13.6{\mu}g/mL$) < ethyl acetate fraction (6.2) < aglycone fraction (2.1). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$ of extract / fractions from Castanea crenata leaf extract / fractions on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activity was in the order: aglycone fraction (0.8) < 50% ethanol extract (0.5) < ethyl acetate fraction (0.3). The scavenging activity ($IC_{50}$ for ${O_2}^{{\cdot}\;-}$ (superoxide anion radical) generated by NBT method was in the order: ethyl acetate fraction (145.5) < aglycone fraction (65.5). The protective effects on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethyl acetate fraction exhibited the most prominent cellular protective effect (${\tau}_{50}$, $191.9{\pm}12.2\;min$ at $10{\mu}g/mL$). The inhibitory effect of aglycone fraction ($9.1{\mu}g/mL$) on elastase was higher than oleanolic and ($13.7{\mu}g/mL$). And the inhibitory effect of aglycone fraction ($21.6{\mu}g/mL$) on tyrosinase was higher than arbutin ($226.2{\mu}g/mL$). But 50% ethanol extract rarely exhibited the inhibitory activity on tryosinase and elastase. Flavonoids were contained in Castanea crenata left (96.3 mg / 100 g dried Castanea crenata leaf). And flavonoids contained in ethyl acetate fraction were kaempferol, quercetin, quercitrin, and so on. Quercitrin is the most abundant component. These results indicate that extract / fractions of Castanea crenata can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging free radical and ROS, Castanea crenata leaf extract/ fractions could be used as new cosmeceutical for whitening and anti-wrinkle products.

Antioxidative and Antiaging Effects of Platycarya strobilacea Extract and Clinical Trial (굴피나무 추출물의 항산화, 항노화 효과 및 인체 시험)

  • Yang, Hee-Jung;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.4
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    • pp.275-286
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    • 2008
  • In this study, we investigated the anti-oxidative, anti-wrinkle and whitening effects of Platycarya strobilacea bark extracts. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$) of extract / fractions of Platycarya strobilacea was in the order: 50% ethanol extract ($6.75{\mu}g/mL$) < deglycosylated aglycone fraction ($6.62{\mu}g/mL$) < ethyl acetate fraction ($4.15{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some Platycarya strobilacea extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activity was ethyl acetate fraction (OSC50, $0.56{\mu}g/mL$) < 50% ethanol extract ($0.02{\mu}g/mL$) < deglycosylated aglycone fraction ($0.01{\mu}g/mL$). The deglycosylated aglycone fraction showed the most prominent scavenging activity. The protective effects of extract / fractions of Platycarya strobilacea on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethanol extract (50%) suppressed photohemolysis in a concentration dependent manner, particularly ethyl acetate fraction exhibited the most prominent cellular protective effect (${\tau}_{50}$, 717.27 min at $10{\mu}g/mL$). The inhibitory effect of Platycarya strobilacea extracts on tyrosinase were investigated to assess their whitening efficacy. Finally, their anti-elastase activities were measured to predict the anti-wrinkle efficacy in the human skin. The inhibitory effect ($IC_{50}$) on tyrosinase of some Platycarya strobilacea extracts was 50% ethanol extract ($243.98{\mu}g/mL$) < ethyl acetate fraction ($153.87{\mu}g/mL$) < deglycosylated aglycone fraction ($137.53{\mu}g/mL$). Also, The inhibitory effect of elastase ($IC_{50}$) of some Platycarya strobilacea extracts was 50% ethanol extract ($31.01{\mu}g/mL$) < ethyl acetate fraction ($14.42{\mu}g/mL$) < deglycosylated aglycone fraction ($1.48{\mu}g/mL$). The cream containing the ethyl acetate fraction of Platycarya strobilacea extracts was formulated. The skin hydration, transepidermal water loss, and the whitening effects were investigated after topical application of the cream. The skin hydration of cream containing extract was increased by $2{\sim}8%$ than the placebo cream, transepidermal water loss was decreased. The cream containing extract suppressed the melanogenesis of skin by 9.55% than the placebo cream. These results indicate that extract / fractions of Platycarya strobilacea can function as antioxidants in biological systems, particularly skin exposed to UV radiation by anti-oxidative activity and protect cellular membranes against ROS. The inhibitory effect on elastase and tyrosinase, and the increase of skin hydration and the whitening effect of the cream containing extract could be applicable to new functional cosmetics for antiaging.

Intracellular Lipid Accumulation Inhibition, Anticancer Activity, and Single Oral Dose Toxicity of Ethanolic Wolfiporia cocos Extracts (에탄올 복령추출물의 지방축적 억제활성, 항암활성 및 단회 경구 독성시험)

  • Park, Na-Hye;Lee, Hwa-Yong;Choi, Jong-Woon;Park, Seung-Chun
    • The Korean Journal of Mycology
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    • v.46 no.3
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    • pp.295-306
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    • 2018
  • In the present study, we compared the effects of 50% ethanolic extracts of Chinese and Korean Wolfiporia cocos (CPE and KPE) on in vitro lipid accumulation in 3T3-L1 cells and their anticancer activities in Sarcoma 180 cells. We further compared the anticancer activities and the 50% inhibitory concentrations ($IC_{50}$) of CPE with KPE with cultivated for one and two years in a landfill and a facility (LPE and FPE), respectively. In addition, the single oral dose toxicities of CPE and KPE were evaluated in mice. Lipid accumulation was inhibited after 48 hours, in CPE and KPE treated 3T3-L1 cells; however, no significant difference was observed between CPE and KPE in their lipid accumulation inhibitory activities. The anticancer activity of KPE was higher than that of CPE at $300{\mu}g/mL$ (p<0.05), revealing the possibility of an auxiliary biological means for origin identification. The anticancer activities of LPE and FPE were significantly stronger than that of CPE (p<0.05) but there was no difference between extracts from one- and two-year-old W. cocos, irrespective of the cultivation method. In single oral dose toxicity tests, CPE and KPE did not induce mortality during the 14-day observation. Thus, the 50% of lethal dose ($LD_{50}$) of CPE and KPE were estimated to be higher than 2,000 mg/kg. Taken together, our results indicate that the anticancer assay could be an auxiliary means of identifying the origin of W. cocos. In addition, artificial cultivation could be an alternative way to reduce the import of W. cocos. Lastly, 50% ethanolic W. cocos extracts could be potential candidates for obesity and cancer managements.

The Role of Heme Oxygenase-1 in Lung Cancer Cells (폐암세포주에서 Heme Oxygenase-1의 역할)

  • Jung, Jong-Hoon;Kim, Hak-Ryul;Kim, Eun-Jung;Hwang, Ki-Eun;Kim, So-Young;Park, Jung-Hyun;Kim, Hwi-Jung;Yang, Sei-Hoon;Jeong, Eun-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.3
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    • pp.304-313
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    • 2006
  • Background : Heme oxygenase-1 (HO-1) is an inducible enzyme that catalyzes the oxidative degradation of heme to form biliverdin, carbon monoxide (CO), and free iron. The current evidence has indicated a critical role of HO-1 in cytoprotection and also in other, more diverse biological functions. It is known that the high expression of HO-1 occurs in various tumors, and that HO-1 has an important role in rapid tumor growth because of its antioxidative and antiapoptotic effects. Therefore, the role of HO-1 was analyzed in human lung cancer cell lines, and especially in the A549 cell line. Material and Methods : Human lung cancer cell lines, i.e., A549, NCI-H23, NCI-H157 and NCI-H460, were used for this study. The expression of HO-1 in the untreated state was defined by Western blotting. ZnPP, which is the specific HO inhibitor we used, and the viability of cells were tested for by conducting MTT assaysy. The HO enzymatic activity, as determined via the bilirubin level, was also indirectly measured. Moreover, the generation of intracellular hydrogen peroxide (H2O2) was monitored fluorimetrically with using a scopoletin-horse radish peroxidase (HRP) assay and 2',7'-dichlorofluorescein diacetate (DCFH-DA). We have also transfected small HO-1 interfering RNA (siRNA) into A549 cells, and the apoptotic effects were evaluated by flow cytometric analysis and Western blotting. Results : The A549 cells had a greater expression of HO-1 than the other cell lines, whereas ZnPP significantly decreased the viability of the A549 cells more than the viability of the other lung cancer cells in a dose-dependant fashion. Consistent with the viability, the HO enzymatic activity also was decreased. Moreover, intracellular H2O2 generation via ZnPP was induced in a dose-dependent manner. Apoptotic events were, then induced in the HO-1 siRNA transfected A549 cells. Conclusion : HO-1 provides new important insights into the possible molecular mechanism of the antitumor therapy in lung cancer.