• Toxicological Research
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• 제17권
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• pp.271-280
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• 2001

We report on the applicability oj a method recently developed in our laboratory for measuring the antioxidant potential of isolated chemicals and extracts derived from natural products. Peroxyl radicals generated by thermal homolysis of 2,2'-azobis-amidinopropane (ABAP) oxidize $\alpha$-keto-${\gamma}$-methiolbutyric acid (KMBA) to ethylene, which is monitored by gas chromatography. Inhibition of ethylene formation in the presence of antioxidants that compete with KMBA for peroxyl radicals is the basis of the Total Oxyradical Scavenging Capacity Assay (TOSCA; Winston et al., 1998). Antioxidative activities of water-soluble extracts of American green tea, the anti-hepatotoxic flavonolignan from milk thistle (Silybum marianum) silybin, Ginkgo biloba extract EGb 761, and a series of naturally occuring sesquiterpene lactones (all ger-macranolides found in in fungi, liverworts, and plants) were studied. The specific TOSC value per $\mu$M silybin was 5.2, which is essentially comparable to that of Trolo $x^{ⓡ}$, a water-soluble vitamine E analog. Tea and Ginkgo extracts exhibited potent peroxyl radical scavenging capacity with values, respectively of =1700 and 1000 $\mu$mols Trolo $x^{ⓡ}$ equivalent per gram dry matter. The known anti-inflammatory activity of some germacranolides prompted study of their antioxidant capacity. None of the lactones exhibited antioxidant capacity toward peroxyl radicals comparable to Trolo $x^{ⓡ}$; costunilide, the most lipophilic, had a TOSC value = to glutathione. The potential role of peroxyl radicals in lipidperoxidation, other cellular damage, and var-ious disease states suggest a possible preventive role for silybin, green tea and Ginkgo biloba in oxidative stress caused by these free radical species.ecies.

• Asian-Australasian Journal of Animal Sciences
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• 제33권7호
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• pp.1113-1125
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• 2020

Objective: This study was investigated the effects of dietary supplementation of Antrodia cinnamomea fermented product on modulation of antioxidation, anti-inflammation, and lipid metabolism in broilers. Methods: Functional compounds and in vitro antioxidant capacity were detected in wheat bran (WB) solid-state fermented by Antrodia cinnamomea for 16 days (FAC). In animal experiment, 400 d-old broiler chickens were allotted into 5 groups fed control diet, and control diet replaced with 5% WB, 10% WB, 5% FAC, and 10% FAC respectively. Growth performance, intestinal microflora, serum antioxidant enzymes and fatty acid profiles in pectoral superficial muscle were measured. Results: Pretreatment with hot water extracted fermented product significantly reduced chicken peripheral blood mononuclear cells death induced by lipopolysaccharide and 2,2'-Azobis(2-amidinopropane) dihydrochloride. Birds received 5% and 10% FAC had higher weight gain than WB groups. Cecal coliform and lactic acid bacteria were diminished and increased respectively while diet replaced with FAC. For FAC supplemented groups, superoxide dismutase (SOD) activity increased at 35 days only, with catalase elevated at 21 and 35 day. Regarding serum lipid parameters, 10% FAC replacement significantly reduced triglyceride and low-density lipoprotein level in chickens. For fatty acid composition in pectoral superficial muscle of 35-d-old chickens, 5% and 10% FAC inclusion had birds with significantly lower saturated fatty acids as compared with 10% WB group. Birds on the 5% FAC diet had a higher degree of unsaturation, followed by 10% FAC, control, 5% WB, and 10% WB. Conclusion: In conclusion, desirable intestinal microflora in chickens obtaining FAC may be attributed to the functional metabolites detected in final fermented product. Moreover, antioxidant effects observed in FAC were plausibly exerted in terms of improved antioxidant enzymes activities, increased unsaturated degree of fatty acids in chicken muscle and better weight gain in FAC inclusion groups, indicating that FAC possesses promising favorable mechanisms worthy to be developed.

• Preventive Nutrition and Food Science
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• 제16권1호
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• pp.12-17
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• 2011

This study was designed to investigate the protective effect of Sasa borealis leaf extract on 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress in LLC-PK1 cells (porcine kidney epithelial cells). The butanol fraction from Sasa borealis leaf extract (SBBF) was used in this study because it possessed strong antioxidant activity and high yield among fractions. Exposure of LLC-PK1 cells to 1 mM AAPH for 24 hr resulted in a significant decrease in cell viability, but SBBF treatment protected LLC-PK1 cells from AAPH-induced cell damage in a dose dependant manner. To determine the protective action of SBBF against AAPH-induced damage of LLC-PK1 cells, we measured the effects of SBBF on lipid peroxidation and antioxidant enzymes activities of AAPH treated cells as well as scavenging activities on superoxide anion radical and hydroxyl radical. SBBF had a protective effect against the AAPH-induced LLC-PK1 cellular damage and decreased lipid peroxidation and increased activities of antioxidant enzymes such as superoxide dismutase and glutathione peroxidase. Furthermore, SBBF showed strong scavenging activity against superoxide anion radical. The $IC_{50}$ value of SBBF was $28.45{\pm}1.28\;{\mu}g/mL$ for superoxide anion radical scavenging activity. The SBBF also had high hydroxyl radical scavenging activity ($IC_{50}=31.09{\pm}3.08\;{\mu}g/mL$). These results indicate that SBBF protects AAPH-induced LLC-PK1 cells damage by inhibiting lipid peroxidation, increasing antioxidant enzyme activities and scavenging free radicals.

• ALGAE
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• 제30권4호
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• pp.313-323
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• 2015

Lipid peroxidation means the oxidative degradation of lipids. The process from the cell membrane lipids in an organism is generated by free radicals, and result in cell damage. Phlorotannins, well-known marine brown algal polyphenols, have been utilized in functional food supplements as well as in medicine supplements to serve a variety of purposes. In this study, we assessed the potential anti-lipid peroxidation activity of phlorofucofuroeckol-A (PFF-A), one of the phlorotannins, isolated from Ecklonia cava by centrifugal partition chromatography in 2,2-azobis (2-amidinopropane) dihydrochloride (AAPH)-stimulated Vero cells and zebrafish system. PFF-A showed the strongest scavenging activity against alkyl radicals of all other reactive oxygen species (ROS) and exhibited a strong protective effect against ROS and a significantly strong inhibited of malondialdehyde in AAPH-stimulated Vero cells. The apoptotic bodies and pro-apoptotic proteins Bax and caspase-3, which were induced by AAPH, were strongly inhibited by PFF-A in a dose-dependent manner and expression of Bcl-xL, an anti-apoptotic protein, was induced. In the AAPH-stimulated zebrafish model, additionally PFF-A significantly inhibited ROS and cell death, as well as exhibited a strong protective effect against lipid peroxidation. Therefore, these results suggest that PFF-A has excellent protective effects against ROS and lipid peroxidation induced by AAPH in both an in vitro Vero cell model and an in vivo zebrafish model.

• Toxicological Research
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• 제26권4호
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• pp.321-327
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• 2010

The ability of 80% ethanol extracts from five medicinal plants, Aralia continentalis, Paeonia suffruticosa, Magnolia denudata, Anemarrhena asphodeloides, and Schizonepeta tenuifolia, to neutralize hydroxyl radical, peroxyl radical and peroxynitrite was examined using the total oxyradical scavenging capacity (TOSC) assay. Peroxyl radical was generated from thermal homolysis of 2,2'-azobis(2-methylpropionamidine) dihydrochloride (ABAP); hydroxyl radical by an iron-ascorbate Fenton reaction; peroxynitrite by spontaneous decomposition of 3-morpholinosydnonimine N-ethylcarbamide (SIN-1). The oxidants generated react with $\alpha$-keto-$\gamma$-methiolbutyric acid (KMBA) to yield ethylene, and the TOSC of the substances tested is quantified from their ability to inhibit ethylene formation. Extracts from P. suffruticosa, M. denudata, and S. tenuifolia were determined to be potent peroxyl radical scavenging agents with a specific TOSC (sTOSC) being at least six-fold greater than that of glutathione (GSH). These three plants also showed sTOSCs toward peroxynitrite markedly greater than sTOSC of GSH, however, only P. suffruticosa revealed a significant hydroxyl radical scavenging capacity. Seven major active constituents isolated from P. suffruticosa, quercetin, (+)-catechin, methyl gallate, gallic acid, benzoic acid, benzoyl paeoniflorin and paeoniflorin, were determined for their antioxidant potential toward peroxynitrite, peroxyl and hydroxyl radicals. Quercetin, (+)-catechin, methyl gallate, and gallic acid exhibited sTOSCs 40~85 times greater than sTOSC of GSH. These four components also showed a peroxynitrite scavenging capacity higher than at least 10-fold of GSH. For antioxidant activity against hydroxyl radical, methyl gallate was greatest followed by gallic acid and quercetin. Further studies need to be conducted to substantiate the significance of scavenging a specific oxidant in the prevention of cellular injury and disease states caused by the reactive free radical species.

• Preventive Nutrition and Food Science
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• 제13권4호
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• pp.259-265
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• 2008

This study was designed to investigate the protective effect of fucoidan against AAPH-induced oxidative stress in LLC-PK1 cells (porcine kidney epithelial cells). Oxidative stress was induced by exposing of LLC-PK1 cells to the 1 mM 2,2'-azobis(2-amidino propane) dihydrochloride (AAPH) for 24 hr. Exposure of LLC-PK1 cells to 1 mM AAPH for 24 hr resulted in a significant (p<0.05) decrease in cell viability, but fucoidan treatment protected LLC-PK1 cells from AAPH-induced cell damage in a dose dependant manner. To investigate the protective action of fucoidan against AAPH-induced damage of LLC-PK1 cells, we measured the effects of fucoidan on lipid peroxidation and antioxidant enzymes activities of AAPH treated cells as well as scavenging activities on superoxide anion radical and hydroxyl radical. Fucoidan had protective effect against the AAPH-induced LLC-PK1 cellular damage and decreased lipid peroxidation and increased activities of antioxidant enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GSH-px). Furthermore, fucoidan showed strong scavenging activity against superoxide anion radical. The $IC_{50}$ value of fucoidan was $48.37{\pm}1.54\;{\mu}g/mL$ for superoxide anion radical scavenging activity. The fucoidan also had high hydroxyl radical scavenging activity ($IC_{50}=32.03\;{\mu}g/mL$). These results indicate that fucoidan protects against AAPH-induced LLC-PK1 cell damage by inhibiting lipid peroxidation, increasing antioxidant enzyme activities and scavenging offree radicals.

• Food Science and Biotechnology
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• 제14권5호
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• pp.586-592
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• 2005

This study was performed to investigate the antioxidant effects of Korean soybean paste extracts (SPE) on 2,2-azobis (2-amidinopropane) dihydrochloride (AAPH)-induced liver damage in rats. Thirty healthy Sprague Dawley rats were selected and divided into 5 groups. Isoflavone contents were measured using HPLC technique. The antioxidant activity was measured in the plasma and liver of the rats with the following results. Levels of isoflavone in fermented soy paste, red pepper paste and soy sauce were 28.9, 30.3 and $3.4\;{\mu}g/g$ for daidzein and 244.3, 187.7 and $6.1\;{\mu}g/g$ for genistein, respectively. The activities of glutamate oxaloacetic transaminase (GOT) and glutamate pyruvate transaminase (GPT) were significantly higher in the AAPH-treated group in the SPE-AAPH group (p<0.05). The thiobarbituric acid reactive substance (TBARS) production was significantly increased in the AAPH-treated liver tissue (P<0.05). Glutathione peroxidase (GPx), glutathione reductase (GR) and catalase in the liver were significantly (p<0.05) decreased by AAPH administration. The glutathione (GSH) concentration was higher in the SPE-treated (Ed- confirm) group than in the control and other groups (p<0.05). These results suggest that SPE led to increased anti oxidative activities against AAPH-induced peroxyl radical.

• 대한한의학회지
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• 제18권1호
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• pp.278-298
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• 1997

This study was performed to prove the antioxidant activities of Cynanchum Wilfordii Hemsley(白何首烏) by way of aqua- acupuncture methods. After 10% & 20% concentrations of liquid extract of Cynanchum Wilfordii Hemsley were acupunctured on Joksamri(足三里) of rats with acute and chronic experimental oxidation by AAPH(2, 2'-azobis(aminoidinopropane), hydrochoride), various kinds of experiments were measured. The results were obtained as follows: 1. In acute antioxidant experiment, the amount of TBARS indicated irregular increase and decrease to over 68% in experiment group in comparison with normal group. 2. In the serum test of acute antioxident experiment, uric acid and total protein and albumin were significantly increased in experiment group in comparison with control group. 3. In chronic antioxident experiment, the change of body weight was significantly decreased in 20% experimental groups in comparison with control group, and only liver significantly increased in 10% & 20% experiment group. 4. In chronic antioxident experiment, total protein significantly increased in 10%, 20% experimental groups and creatinine and BUN were significantly decreased in 10% experiment group. 5. In the chronic antioxident experiment, serum GOT, GPT activity didn't show no significantly change, LDH activity was significantly increase in 10% experiment group in comparison with control group. 6. In the chronic antioxident experiment, the tolerance test with enzyme activities of erythrocyte membrane was significantly decreased in 10% & 20% experimental groups. 7. In the chronic antioxident experiment, the change of TBA was significantly decreased in 10% & 20% experimental group. 8. In the chronic antioxident experiment, the amount of cytochrome p-450 was significantly increased in 10% & 20% experimental groups in comparison with control group. 9. In the chronic antioxident experiment, the amount of cytochrome b5 was significantly increase in 20% experimental group in comparison with control group. 10. In the chronic antioxident experiment, the change of activity of superoxide dismutase, and catalase were significantly increased in 20% experiment group in comparison with control group. From the above result, it was concluded considers the liquid extract of Cynanchum Wilfordii Hemsley(白何首烏) could be used as antiaging drugs, because it delayed the aging process and retarded senility.

• 생약학회지
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• 제43권1호
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• pp.66-71
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• 2012

Cirsium japonicum var. ussuriense is often used in treatment of human disease such as hemorrhage, blood congestion and inflammation. This study was accomplished to evaluate the antioxidant properties of the leaf (CLE) and flower (CFE) extracts of C. japonicum var. ussuriense to protect normal human red blood cells (RBC) and plasma samples against oxidative damage in vitro. CLE and CFE were prepared by extracting with hot water. In red blood cells and plasma, oxidative hemolysis and lipid peroxidation induced by the aqueous peroxyl radical generator [2,2'-azobis (2-amidinopropane) dihydrochloride, AAPH] were significantly suppressed by CLE or CFE in a dose-dependent manner at the same time. CLE and CFE also prevented the depletion of cytosolic antioxidant glutathione (GSH) in RBC. These results suggest that the leaves and flowers of C. japonicum var. ussuriense may have the antioxidant properties.

• 한방비만학회지
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• 제22권1호
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• pp.1-10
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• 2022

Objectives: To investigate the protective effect of hesperidin and hesperetin against oxidative stress in 2,2'-azobis (2-aminopropane) dihydrochloride (AAPH)-induced liver toxicity in rats. Methods: Hesperidin or hesperetin (200 mg/kg/day, respectively) was orally administered for 7 days once daily in rats. Subsequently, AAPH (50 mg/kg/day) was administered intraperitoneally. Lipid peroxidation, nitric oxide production, catalase activity, and protein expressions of nuclear factor-kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) in the liver tissues were measured. Results: Administration of hesperidin and hesperetin significantly decreased serum aspartate transaminase and alanine transaminase levels in AAPH-induced oxidative stress liver tissues compared with control group. Lipid peroxidation and nitric oxide (NO) production were also significantly reduced by hesperidin and hesperetin in AAPH-induced oxidative stress liver tissues. In particular, lipid peroxidation levels of hesperetin-administered group significantly decreased to 5.02 nmole/mg protein in oxidative stress rats. Hesperidin and hesperetin significantly increased antioxidant activity, such as that of catalase. Furthermore, administration of hesperidin and hesperetin substantially down-regulated the expression of NF-κB and iNOS in liver tissues. Administration of hesperidin reduced NO levels and iNOS expression more than in the hesperetin-administered group. Conclusions: Administration of hesperidin and hesperetin led to a reduction in AAPH-induced liver toxicity by regulating oxidative stress.