• Asian Pacific Journal of Cancer Prevention
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• v.16 no.5
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• pp.1833-1837
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• 2015

An aristolactam-type alkaloid, isolated from Orophea enterocarpa, is enterocarpam-III (10-amino-2,3,4,6-tetramethoxyphenanthrene-1-carboxylic acid lactam). It is cytotoxic to various human and murine cancer cell lines; however, the molecular mechanisms remain unclear. The aims of this study were to investigate cytotoxic effects on and mechanism (s) of human cancer cell death in human hepatocellular carcinoma HepG2 and human invasive breast cancer MDA-MB-231 cells compared to normal murine fibroblast NIH3T3 cells. Cell viability was determined by MTT assay to determine $IC_{10}$, $IC_{20}$ and $IC_{50}$ levels, reactive oxygen species (ROS) production with 2',7'-dichlorohydrofluorescein diacetate and the caspase-3, -8 and -9 activities using specific chromogenic (p-nitroaniline) tetrapeptide substrates, viz., DEVD-NA, IETD-NA and LEHD-NA and employing a microplate reader. Mitochondrial transmembrane potential (MTP) was measured by staining with 3, 3'-dihexyloxacarbocyanine iodide ($DiOC_6$) and using flow cytometry. The compound was cytotoxic to HepG2 and MDA-MB-231 cells with the $IC_{50}$ levels of $26.0{\pm}4.45$ and $51.3{\pm}2.05{\mu}M$, respectively. For murine normal fibroblast NIH3T3 cells, the $IC_{50}$ concentration was $81.3{\pm}10.1{\mu}M$. ROS production was reduced in a dose-response manner in HepG2 cells. The caspase-9 and -3 activities increased in a concentration-dependent manner, whereas caspase-8 activity did not alter, indicating the intrinsic pathway activation. Enterocarpam-III decreased the mitochondrial transmembrane potential (MTP) dose-dependently in HepG2 cells, suggesting that the compound induced HepG2 cell apoptosis via the mitochondrial pathway. In conclusion, enterocarpam-III inhibited HepG2 and MDA-MB-231 cell proliferation and induced human HepG2 cells to undergo apoptosis via the intrinsic (mitochondrial) pathway and induction of caspase-9 activity.

• Journal of Food Hygiene and Safety
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• v.30 no.4
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• pp.383-389
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• 2015

This study was to examine the antioxidative activity of fermented Rhynchosia nulubilis (FRN) in obese rats. Oxidative stress due to reactive oxygen species (ROS) can cause oxidative damage to cells. Mitochondria are especially important in the oxidative stress as ROS have been found to be constantly generated as an endogen threat. Mitochondrial defense depends mainly on superoxide dismutase whereas microsomal defense depends on catalase, which is an enzyme abundant in microsomes. Seven weeks-aged female Sprague-Dawley rats were divided into four groups and fed high fat diets for 44 days. Also fermented Rhynchosia nulubilis was administered orally for 44 days at 7.5 ml/kg of body weight of rats. The antioxidative activities of fermented Rhynchosia nulubilis were measured by the superoxide dismutase, catalase, malondialdehyde levels in liver homogenate. The levels of malondialdehyde in FRN-treated groups were lower than those in obese groups. Superoxide dismutase and catalase levels were significantly increased. These results demonstrated that fermented Rhynchosia nulubilis had the inhibitive effects of oxidative stress in obese rats, suggesting that fermented Rhynchosia nulubilis would be used as an ingredient of the useful functional products.

• Journal of Pharmacopuncture
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• v.10 no.3
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• pp.53-62
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• 2007

Objective The objective of this study was to investigate the antioxidative effects of Puerariae Radix extract. Method Total antioxidant capacity (TAC), Total antioxidant response (TAR), Total phenolic content, Reactive oxygen species (ROS), 1,1-Diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activities, lipid peroxidation were examined. Result Total antioxidant status was examined by total antioxidant capacity(TAC) and total antioxidant response(TAR) against potent free radical reactions. TAC and TAR of Puerariae Radix extract at the concentration of 5 mg/ml were 2.02 and 1.50 mM Trolox equivalents, respectively. Total phenolic content of Puerariae Radix extract at the concentration of 5 mg/ml was 2.29 mM gallic acid equivalent. Concentration of Puerariae Radix extract at which DPPH radical scavenging activity was inhibited by 50% was 5.91 mg/ml as compared to 100% by pyrogallol solution as a reference. The inhibitory effect of the extract on lipid peroxidation was examined using rat liver mitochondria induced by FeSO4/ascorbic acid. Puerariae Radix extract at the concentration of 1 mg/ml slightly but significantly decreased TBARS concentration. The extract further prevented lipid peroxidation in a dose-dependent manner. The effect of Puerariae Radix extract on reactive oxygen species (ROS) generation was examined using cell-free system induced by hydrogen peroxide/FeSO4. Addition of 1 mg/ml of Puerariae Radix extract significantly reduced dichloroflurescein (DCF) fluorescence. The extract caused concentration-dependent attenuation of the increase in DCF fluorescence, indicating that the extract significantly prevented ROS generation in vitro. Thus antioxidant effects of Puerariae Radix extract seem to be due to, at least in part, the prevention from free radicals-induced oxidation, followed by inhibition of lipid peroxidation. Conclusion As a result, Puerariae Radix seems to have antioxitative effect and antioxidant compount.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.9
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• pp.4367-4375
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• 2016

The objective of the present study was to evaluate the anticancer and radio-sensitizing efficacy of a Withania somnifera extract/Gadolinium III oxide nanocomposite (WSGNC) in mice. WSGNC was injected to solid Ehrlich carcinoma-bearing mice via i.p. (227 mg/kg body weight) 3 times/week during 3 weeks. Irradiation was performed by whole body fractionated exposure to 6Gy, applied in 3 doses of 2 Gy/week over 3 weeks. Biochemical analyses as well as DNA fragmentation were performed. Treatment of solid Ehrlich carcinoma bearing mice with WSGNC combined with ${\gamma}$-radiation led to a significant decrease in the tumor size and weight associated with a significant decrease in mitochondrial enzyme activities, GSH content and SOD activity as well as a significant increase in caspase-3 activity, MDA concentration and DNA fragmentation in cancer tissues. Combined treatment of WSGNC and low dose of ${\gamma}$-radiation showed great amelioration in lipid peroxidation and antioxidant status (GSH content and SOD activity) in liver tissues in animals bearing tumors. It is concluded that WSGNC can be considered as a radio-sensitizer and anticancer modulator, suggesting a possible role in reducing the radiation exposure dose during radiotherapy.

• Journal of the Korean Society of Food Science and Nutrition
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• v.13 no.4
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• pp.459-468
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• 1984

The currently accepted model of membrane structure proposes a dynamic, asymmetric lipid matrix of phospholipids and cholesterol with globular proteins embedded across the membrane to various degrees. Most phospholipids are in the bilayer arrangement and also closely associated with integral membrane proteins or loosely associated with peripheral proteins. Biological functions of membrane, such as membrane-bound enzyme functions and transport systems, are influenced by the membrane physical properties, which are determined by fatty acid composition of phospholipids, polar head group composition and membrane cholesterol content. Polar and non-polar region of the phospholipid molecule can interact, with changes in the conformation of a membrane-associated protein altering either its catalytic activity or the protein's interaction with other membrane proteins. Mammalian dietary studies attempted to change the lipid composition of a few cell membranes have shown comparisons, using essential fatty acid-deficient diets. In recent years, Clandinin and a few other workers have pioneered the study proving the influence of dietary fat fed in a nutritionally complete diet on composition of phospholipid classes of cell membrane. Modulation caused by diet fat was rapid and reversible in phospholipid fatty acyl composition of membranes of cardiac mitochondria, liver cell, brain synaptosome and lymphocytes. These changes were at the same time, accompanied by variety of membrane associated functions controlled by membrane-bound enzymes, tranporter and receptor proteins. The findings suggest the basic concept of the necessity of dietary fatty acid balance if consistency of optimal membrane structural lipid composition is to be maintained, as well as the overall inadequacy of describing the nutritional-biochemical quality of a dietary fat solely by its content of linoleic acid. Furthermore, they give light on the possible application to clinical and preventive medicine.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.45 no.6
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• pp.553-560
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• 2012

Hizikia fusiformis, a brown alga that is widely consumed in Korea, Japan, and China, possesses a number of potentially beneficial compounds, including antioxidants and anticoagulants. However, the molecular mechanisms of H. fusiformis in hepatoma cells have not been elucidated. This study investigated the antiproliferative effect and mechanism of action of a glycoprotein from H. fusiformis (HFGP) in HepG2 human hepatoma cells. In an MTS assay, 25 ${\mu}g/mL$ HFGP inhibited the proliferation of HepG2 cells by $52.36{\pm}2.37%$. HFGP caused the dose-dependent growth inhibition of HepG2 cells by inducing apoptosis and a sub-G1 phase arrest. The antiproliferative activity of HFGP was confirmed based on the expression of several apoptosis-related proteins, which was assessed by Western blot analysis. The expressions of Fas, Fas-associated death domain protein, Bax, and Bad was significantly up-regulated in HFGP-treated cells, and HFGP induced the translocation of Bax to mitochondria and the release of cytochrome c into the cytosol. Therefore, HFGP might be useful in the treatment of liver cancer.

• Food Science and Biotechnology
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• v.16 no.4
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• pp.531-536
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• 2007

The aerial part of Artemisia iwayomogi Kitamura has traditionally been used for inflammation, infectious disease, cancer, pyretic, diuretic, liver protective effect, and choleretic purposes in Korea. We investigated that the essential oil induces apoptosis in KB cell as evidenced by Hoechst-33258 dye staining, flow cytometry (cell cycles), and DNA fragmentation for nuclear condensation and Western blotting for activation of caspases-3, -8, -9, Bax, Bcl-2, cytochrome c, and poly (ADP-ribose) polymerase (PARP) cleavage. In the present study, we found that the essential oil could induce apoptosis in KB cells, as characterized by DNA fragmentation, activation of caspase-3, -8, and -9, and PARP cleavage. The efficacious induction of apoptosis was observed as a dose-dependent. The essential oil-induced apoptotic cell death was accompanied by up-regulation of Bax and down-regulation of Bcl-2. The essential oil also caused the loss of mitochondrial membrane potential and cytochrome c release from mitochondria to cytosol. These findings indicate that mitochondrial pathways might be involved in the essential oil-induced apoptosis and enhance our understanding of the anticancer function of the essential oil in herbal medicine.

• Korean Journal of Plant Resources
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• v.19 no.6
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• pp.649-654
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• 2006

The objective of present study was to investigate the anti oxidative and hepatoprotective effects of tomato extracts. Total antioxidant capacity and total antioxidant response were 5.5 and $19.8{\mu}g$ Trolox equivalent per mg of tomato extract, respectively. DPPH radical scavenging activity of tomato extracts ($10mg\;ml^{-1}$) was 70% as compared to 100% by pyrogallol solution as a reference. The effect of the tomato extracts on lipid peroxidation was examined using rat liver mitochondria induced by iron/ascorbate. Tomato extracts at the concentration of $0.5mg\;ml^{-1}$ significantly decreased TBARS concentration. Tomato extracts prevented lipid peroxidation in a dose-dependent manner. The effect of the tomato extracts on reactive oxygen species (ROS) generation was examined using cell-free system induced by $H_2O_2/FeSO_4$. Addition of $1mg\;ml^{-1}$ of tomato extracts significantly reduced dichlorofluorescein (DCF) fluorescence. Tomato extracts caused concentration-dependent attenuation of the increase in DCF fluorescence, indicating that tomato extracts significantly prevented ROS generation in vitro. The effect of tomato extracts on cell viability and proliferation was examined using hepatocyte culture. Primary cultures of rat hepatocytes were incubated with 1mM tert-butyl hydroperoxide (t-BHP) for 90 min in the presence or absence of tomato extracts. MTT values by addition of tomato extracts at the concentration of 2, 10, and $20mg\;ml^{-1}$ in the presence of t-BHP were 13, 33 and 48%, respectively, compared to 100% as control. Tomato extracts increased cell viability in a dose-dependent manner. These results demonstrate that tomato extracts suppressed lipid peroxidation and t-BHP-induced hepatotoxicity and scavenged ROS generation. Thus antioxidant and hepatoprotective effects of tomato extracts seem to be due to, at least in part, the prevention from free radicals-induced oxidation, followed by inhibition of lipid peroxidation.

• The Journal of Korean Medicine
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• v.28 no.1 s.69
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• pp.137-147
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• 2007

Objective : The objective of this study was to investigate the antioxidative effects of Carthami Flos extract. Methods : Total antioxidant status was examined by total antioxidant capacity(TAC) and total antioxidant response(TAR) against potent free radical reactions. The effect of Carthami Flos extract was examined far details of total phenolic content concentration at which 1,1-dipheny1-2-picrylhydrazyl(DPPH) radical scavenging activity was inhibited, the inhibitory effect on lipid peroxidation, and the effect on reactive oxygen species(ROS) generation. Results : TAC of Carthami Flos extract at the concentration of 5 mg/ml was 1.84 mM Trolox equivalent. 2. TAR of Carthami Flos extract, on the other hand, couldn't be determined due to interference from unidentified compounds. 3. Total phenolic content of Carthami Flos extract at the concentration of 5 mg/ml was 2.01 mM gallic acid equivalent. 4. Concentration of Carthami Flos extract at which DPPH radical scavenging activity was inhibited by 50% was 6.43 mg/ml as compared to 100% by Pyrogallol solution as a reference. 5. The inhibitory effect of the extract on lipid peroxidation was examined using rat liver mitochondria induced by FeS04/ascorbic acid. Carthami Flos extract at the concentration of 10 ms/ml slightly but significantly decreased TBARS concentration. The extract continued to prevent lipid peroxidation in a dose-dependent manner. 6. The effect of Carthami Flos extract on reactive oxygen species(ROS) generation was examined using a cell-free system induced by hydrogen peroxide/FeS04. Addition of 1 mg/ml of Carthami Flos extract significantly reduced dichlorofluorescein(DCF) fluorescence. Carthami Flos extract caused concentration-dependent attenuation of the increase in DCF fluorescence, indicating that the ektract significantly prevented ROS generation in vitro. Conclusion: : Antioxidant efffcts of Carffami ffor extract seem to be due, at least in part, to the prevention offree radical-induced oxidation, fellowed by inhibition of lipid peroxidation.

• The Journal of Korean Medicine
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• v.28 no.1 s.69
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• pp.148-158
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• 2007

Objective : The objective of this study was to investigate the antioxidant effects of Mori Folium extract. Methods Total antioxidant status was examined by total antioxidant capacity(TAC) and total antioxidant response(TAR) against potent free radical reactions. The effect of Mori Folium extract was examined by measuring total phenolic content, concentration at which 1,1-dipheny1-2-picrylhydrazyl(DPPH) radical scavenging activity was inhibited, inhibitory effect on lipid peroxidation, and the effect on reactive oxygen species(ROS) generation. Results : 1. TAC and TAR of Mori Folium extract at the concentration of 5 mg/ml were 1.61 and 1.24 mM Trolox equivalents, respectively. 2. Total phenolic content of Mori Folium extract at the concentration of 5 mg/Ml was 1.70 mM gallic acid equivalent. 3. Concentration of Mori Folium extract at which DPPH radical scavenging activity was inhibited by 50% was 2.29 m9/m4 as compared to 100% by Pyrogallol solution as a reference. 4. The inhibitory effect of the extract on lipid peroxidation was examined using rat liver mitochondria induced by FeSO$_4$/ascorbic acid. Mori Folium extract at the concentration of 10 mg/ml significantly decreased thiobarbituric acid reactive substances(TBARS) concentration. The extract prevented lipid peroxidation in a dose-dependent 5. The effect of Mori Folium extract on reactive oxygen species(ROS) generation was examined using a celt-free system induced by hydrogen peroxide FeSO$_4$. Addition of 1 mg/ml of Mori Folium extract significantly reduced dichlorofluorescein(DCf) fluorescence. The extract caused concentration-dependent attenuation of the increase in DCF fluorescence, indicating that the extract significantly prevented ROS generation in vitro. Conclusion ; The antioxidant effects of Mori Folium extract seem to be due, at least in part, to the prevention offree radical-induced oxidation, fllowed by inhibition of lipid peroxidation.