• Title/Summary/Keyword: DPPH assay

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In vivo and In vitro Anti-lipid Peroxidative Effect of the Extract Complex of Korean Anti-thirst Drugs (한방 소갈약 추출물 및 복합물에 의한 In vivo 및 In vitro 지질과산화 저해효과)

  • 이경태;박동영;박희준;정현주;박건영;최종원
    • YAKHAK HOEJI
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    • v.46 no.5
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    • pp.358-363
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    • 2002
  • In Oriental medicine, the prescriptions composed of several herb medicines have been used. It is still unclear how the sum of several extracts of anti-thirst drugs represents the anti-lipid peroxidative action. Three anti-thirst herb medicines, Kalopanax pictus (K), Pueraria thunbergiana (P) and Rhus verniciflua (R), were extracted with MeOH and $H_2O$, respectively, and the former one was fractionated into the resultant EtOAc extract. Each extract was reconstituted to give KPR311, KPR131 and KPR113 where, for example, KPR311 represents the complex of K-P-R {3:1:1 (w/w/w)} of the three extracts. The order of the inhibitory effect in bromobenzene-induced lipid peroxidation in rats was as follows: EtOAc extract>$H_2O$ extract>MeOH extract. Extract complexes were found to be more potent than the extracts of individual crude drugs. The KPR131 of EtOAc extract was found to be the most potent among the tested samples. These anti-lipid peroxidative effects were also supported by the decrease of aniline hydroxylase activity and aminopyrine N-demethylase activity, on the other hand by the increase of epoxide hydrolase activity. All the tested samples were assayed in vitro antioxidative effects such as DPPH assay, ADP/NADPH/Fe$^{3+}$ assay and ascorbic acid/Fe$^{2+}$ assay. The EtOAc extracts also showed the most significant antioxidative effects. These results suggest that the sum of anti-thirst drugs could reflect the effects of respective crude drugs.s.s.

Anti-oxidant Effects of Samultang-Gami on MEF Cells (사물탕가미방(四物湯加味方)의 항산화 활성에 대한 실험적 연구)

  • Jung, Jae-Joong;Goo, Sun-Young;Go, Eun-Bi;Sung, Jung-Suk;Kim, Dong-Il
    • The Journal of Korean Obstetrics and Gynecology
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    • v.23 no.3
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    • pp.26-37
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    • 2010
  • Purpose: This experiment is designed to find out anti-oxidant effects of Samultang-Gami which was composed of Rehmanniae Radix(RR), Angelicae Gigantis Radix, Cnidii Rhizoma(CR), Paeoniae Radix(PR), Cortex Moutan Radicis, Hedyotis Diffusa(HD) and Caesalpinia Sappan on MEF cells. Methods: In vitro antioxidant effects were measured by MTT assay, DPPH assay, cell cycle analysis, AnnexinV-FITC/PI assay and DAPI staining using MEF cells treated with various concentrations of 70% ethanol extract of Samultang-Gami. Results: 1. In the scavenging for DPPH radical, the each treated groups of PR, CR and HD showed positive effects. RR and CR increased the viability of oxidative damaged MEF cells in a dose-dependent manner. 2. 70% ethanol extract of Samultang-Gami was shown best antioxidative effect in the concentration of $0.5mg/m{\ell}$. 3. The treatment of Samultang-Gami in oxidative damaged MEF cells didn't have any effect on cell cycle restoration. but it could lower late apoptosis rate a little and be observed the protection of nucleus. Conclusion: It can be concluded that Samultang-Gami, RR and CR have antioxidant effects on MEF cells.

Antioxidative Activities and Inhibition Effects on Oxidative DNA Damage of Valeriana fauriei (쥐오줌풀의 항산화 및 산화적 DNA 손상 억제 활성)

  • Park, Jae Ho;Jang, Tae Won;Lee, Seung Hyun
    • Korean Journal of Medicinal Crop Science
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    • v.24 no.6
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    • pp.464-470
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    • 2016
  • Background: Valeriana fauriei (Valerianaceae) has been used to as a traditional medicine to treat a variety of symptoms, including headache, insomnia, hypertension, and menstrual irregularity. However, the present study investigates the species' antioxidant activity and its inhibition of oxidative DNA damage, which have yet to be studied. Methods and Results: The antioxidant activity was assessed using radical scavenging assays with 1,1-diphenyl-2-picryl hydrazyl (DPPH) and, 2, 2'-azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) and a reducing power assay. The total phenol content was also analyzed, and phenolic compounds were detected using HPLC/UV, whereas the inhibitory effect of Valeriana fauriei on oxidative DNA damage was measured using ${\phi}-174$ RF I plasmid DNA cleavage assay. The DPPH and ABTS radical scavenging activity were $75.17{\pm}3.55%$ and $95.83{\pm}0.63%$, repectively, and the reducing power was $93.14{\pm}1.74$ at $200{\mu}g/m{\ell}$. The total phenol content was $10.24{\pm}0.04mg/g$, whereas chlorogenic acid, catechin, caffeic acid and epicatechin were identified using HPLC/UV, and the ${\phi}-174$ RF I plasmid DNA cleavage assay indicated that V. fauriei provided protection against oxidative damage. Conclusions: The results of the present study suggest that V. fauriei has powerful antioxidant activity that can provide protective effects against the oxidative DNA damage caused by free radicals. The species, therefore, provides a valuable resource for the development of natural pharmaceutical to treat aging, cancer, and degenerative diseases.

Ethanol Extract of Forsythiae Fructus Inhibits the Production of Inflammatory Mediators in LPS-stimulated BV-2 Microglial Cells (연교 추출물의 Microglia에서 LPS에 의해 유도되는 염증매개물질 생성 억제 효과)

  • Kim, Sung-Yun;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.93-102
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    • 2008
  • Objectives : Forsythiae Fructus (Forsythia koreana Nakai) has been used anti-inflammatory, diuretics, antidote, and antibacterials in traditional herbal medicine. The present study is focused on the inhibitory effect of Forsythiae Fructus ethanol extract (FF-E) on the production of inflammatory mediators such as NO, iNOS and proinflammatory cytokines ($TNF-{\alpha}$, $IL-1{\beta}$ and IL-6) in LPS-stimulated BV-2 cells, a mouse microglial cell line, and investigated the scavenging activity of FF-E. Methods : BV-2 cells were pre-incubated with FF-E for 30 min and then stimulated with LPS (1 ${\mu}g/m{\ell}$) at indicated times. Cell toxicity of GCF was determined by MTT assay. The levels of NO, PGE2 and cytokines were measured by Griess assay and ELISA. The mRNA and protein expressions of iNOS and cytokines were determined by RT-PCR and Western blotting. Free radical scavenging activity of GCF was determined by DPPH assay in tube test. Results : FF-E significantly inhibited the excessive production of NO, $PGE_2$, $TNF-{\alpha}$, and $IL-1{\beta}$ in LPS-stimulated BV-2 cells. In addition, FF-E attenuated the mRNA and protein expressions of iNOS, and proinflammatory cytokines. FF-E also significantly scavenged the DPPH free radicals in a dose-dependent manner. Conclusions : These results indicate that FF-E exhibits anti-inflammatory property by suppressing the transcription of inflammatory mediator genes, suggesting the anti-inflammatory property of FF-E may make it useful as a therapeutic agent for the treatment of human neurodegenerative diseases.

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Antioxidative and Anti-inflammatory Effect of Ethanol Extract from Duchesnea chrysantha (사매 에탄올 추출물의 항산화 및 항염증 효과)

  • Lee, Deok-Jae;Jeon, In-Hwa;Kim, Hyeon-Soo;Cho, Il-Young;Jang, Seon-Il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.1
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    • pp.59-66
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    • 2012
  • Oxidative stress has been implicated in cutaneous damage in various inflammatory skin diseases, including atopic dermatitis. The present study was undertaken to investigate the antioxidative and anti-inflammatory activities of the extract of Duchesnea chrysantha (DCE). DEC was prepared by extracting with 80% ethanol. Total flavonoids and polyphenols were measured by a colorimetric assay. The free radical scavenging activity of the extract was analyzed by the DPPH (1,1-diphenyl-2-picryl hydrazyl), ABTS (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) and Griess reagent assay. An oxidative product of nitric oxide (NO), was measured in the culture medium by the Griess reaction. The level of prostaglandin $E_2$ ($PGE_2$) was measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were measured by Western blot analysis. Total flavonoid and polyphenol contents of DCE were included $24.73{\pm}0.45$ and $178.77{\pm}2.65$, respectively. DCE significantly increased electron donating ability (DPPH), nitrite scavenging (NO) and ABTS reducing activity in dose dependant. We investigated the anti-inflammatory effects of DCE on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. DCE significantly suppressed NO and prdstaglandin $E_2$ ($PGE_2$) in dose dependant. Furthermore, the levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with DCE in a dose dependent manner. These results suggest that DEC may has value as natural product with its high quality functional components, antioxidative and anti-inflammatory activities.

Antioxidant Activity of Glycyrrhiza cultivar Extracts

  • Kim, Minhee;Kang, Myunghoon;Kim, Wonnam
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.88-88
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    • 2019
  • Antioxidants are involved in the defense mechanism against the attack of free radicals. This study was carried out to determine the antioxidant activity of new variety of Glycyrrhiza cultivar radix, Wongam and Sinwongam. Dissolved freeze dried Wongam and Sinwongam extracts were filtered by $0.2{\mu}m$ filter and serially diluted at the concentrations of $10{\mu}g/mL$, $50{\mu}g/mL$, $100{\mu}g/mL$, $500{\mu}g/mL$, and $1000{\mu}g/mL$. The antioxidant potential was determined by DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity, ABTS (2,2-azino-bis (3-rthylbenzthiazoline-6-sulfonic acid) diammonium salt) radical cation decolorization assay, nitrite radical scavenging assay, and ferric reducing antioxidant power (FRAP) assay. DPPH radical scavenging activities (i.e. the highest value $50.9{\pm}0.8%$ by Wongam and $82.6{\pm}1.1%$ by Sinwongam), ABTS radical scavenging activities (i.e. the highest value $88.1{\pm}1.8%$ by Wongam and $98.6{\pm}0.1%$ by Sinwongam), and nitrite radical scavenging activities (i.e. the highest value $87.3{\pm}1.6%$ by Wongam and $89.8{\pm}0.8%$ by Sinwongam) increased in a dose-dependent manner. In addition, ferric reducing power activities also increased in a dose-dependent manner. The FRAP value of Wongam and Sinwongam extracts were $0.72{\pm}0.03$ and $0.99{\pm}0.06$ compared to ascorbic acid, as a positive control, was $1.32{\pm}0.02$. These results suggested that Wongam and Sinwongam have beneficial effects as a potent antioxidant.

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In vitro Antioxidant Activity of the Aqueous of Angelicae gigas Nakai Leaves (당귀 잎의 항산화 활성)

  • Park, Sung-Jin;Yoon, Jung-Han;Kim, Young-Eon;Yoon, Won-Byong;Kim, Jong-Dai
    • Food Science and Preservation
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    • v.18 no.6
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    • pp.817-823
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    • 2011
  • Angelicae gigas Nakai has been used as a traditional medicine as well as an edible vegetable in South Korea. In this study, the total phenolic and flavonoid content and antioxidants of A. gigas Nakai leaves were examined in vitro via hydroxyl-radical-scavenging activity, reducing power activity, metal chelating assay, and DPPH-free -radical-scavenging assay. Among all the extracts from A. gigas Nakai leaves, the ethanol extract showed the strongest effects in all of the assays. The $EC_{50}$ values for the DPPH-radical-scavenging activities of ethanol, methanol, and water extracts were 31.47, 42.14, and $58.47{\mu}g/mL$, respectively. Among the extracts from A. gigas Kakai leaves, the ethanol extract had the highest levels of total phenolics ($7.84{\pm}1.46$ mg TAN/g) and total flavonoids ($4.23{\pm}0.03$ mg QE/g), which correlated strongly with the individual phenolic-compound (p-hydroxybenzoic acid, vanillin, and trans-ferulic acid) contents. The ethanol extract also showed stronger antioxidant activity than tocopherol in hydroxyl- radical-scavenging activity assay. These results indicate that the ethanol extract of A. gigas Kakai leaves possesses significant antioxidant properties, which suggests its great potential as a functional-food ingredient in the food industry.

Anti-oxidant, Anti-aging, and Whitening Effects of Viscum album var. coloratum In Vitro (한국산 겨우살이의 항산화, 항노화 및 미백 효과)

  • Chang-Eui, Hong;Su-Yun, Lyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.4
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    • pp.295-302
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    • 2022
  • In this study, we investigated the anti-oxidant, anti-aging, and skin whitening effects of Korean mistletoe (Viscum album var. coloratum). The mistletoe fraction was composed of four types: hexane (HX), ethyl acetate (EA), butanol (BU), and methylene chloride (MC). In total phenol content assay, HX showed the highest phenol content among four fractions. In addition, EA significantly increased 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and catalase-like activities, and MC significantly increased superoxide dismutase (SOD)-like activity. When we compared IC50 value in the hyaluronidase and elastase inhibition assay, MC had the lowest IC50. In addition, we also performed tyrosinase inhibition assay to demonstrate the possibility of Korean mistletoe as a cosmetic component. HX showed the highest tyrosinase inhibition rate among the fractions.

The Antioxidant Activities and Neuroprotective Effects of Hot Water Extracts from Torreyae Semen (비자 열수 추출물의 항산화 활성 및 뇌신경세포 보호효과 연구)

  • Lee, Soong-In;Choi, Chan-Hun;Kim, Jeong-Sang;Lim, Seong-Soo;Jung, Hyun-Woo
    • The Korea Journal of Herbology
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    • v.32 no.6
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    • pp.41-48
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    • 2017
  • Objectives : This study was designed to estimate the antioxidative and neuroprotective effects of Torreyae Semen hot water extracts (TS). Methods : Torreyae Semen was extracted by hot water for 2 hours with a temperature of 105 degrees. Polyphenols and total flavonoid were measured and LC-MS/MS was used to certificate anticipated antioxidative compounds. The antioxidant activities of TS were measured as scavenging effects of 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and Nitrite Oxides (NO). Cell viability and proliferation rate was measured MTT assay. The toxicities to thymocytes and splenocytes were evaluated by the proliferation rate of primary cultured cells of 7 weeks, male Balb/c mice. The antioxidant activities of TS on C6 mouse glioma cells were measured by the analysis of total glutathione contents variation. The neuroprotective effects against oxidative stresses were measured by MTT assay. Results : Polyphenols of TS was $92.00{\pm}1.24{\mu}g/mg$, and total flavonoids was $0.36{\pm}0.14{\mu}g/mg$. TS includes gallocatechin, epigallocatechin, gallocatechin gallate and epigallocatechin gallate. TS included gallocatechin, epigallocatechin, gallocatechin gallate, epigallocatechin gallate. TS showed DPPH and NO scavenging effects as dose-dependent manner at the concentrations of $0-10mg/m{\ell}$. In MTT assay, TS shows no significant toxicity to C6 cells, primary cultured thymocytes and splenocytes of Balb/c mice. TS increased the level of total glutathiones. TS increased cell viabilities of C6 cells against oxidative stresses such as $H_2O_2$, sodium nitroprusside (SNP), Rotenone at the concentrations of $0-0.063mg/m{\ell}$. Conclusions : TS shows the antioxidant and neuroprotecitive effects in these experiments.

Antioxidant Effects of PanaX ginseng in Mouse GC-1 Spennatogonia Cells (인삼(人蔘)이 생쥐의 남성 생식세포 GC-1 spermatogonia의 항산화에 미치는 영향)

  • Shim, Kyung-Jun;Kang, Ji-Ung;Choi, Bong-Jae;Park, Soo-yeon;Chang, Mun-Seog;Park, Seong-Kyu
    • The Korea Journal of Herbology
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    • v.24 no.2
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    • pp.93-98
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    • 2009
  • Objectives : Previously we reported that the roots of Panax ginseng C.A. Meyer (Araliaceae) increased sperm count and motility. also induced spermatogenesis via cAMP-responsive element modulator(CREM) activation in rat testes. In this study, for the first step of spermatogenesis in germ cell lines, the antioxidant activity of Panax ginseng were examined in mouse GC-1 spermatogonia cells. Methods : The extract was studied on diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity, GC-1 cell viability by a modified MIT assay. H202-induced cytotoxicity by MIT assay and lipid peroxidation by malondialdehyde (MDA) formation. respectively. Results: The results showed that the extract scavenged DPPH radical with the IC50 being 0.631 mg/mi. The extract at concentrations of 5, and 10, 50, 100, 250 ${\mu}$g/mi increased GC-1 cell viability significantly(p < 0.05, and p < O.O1). Hydrogen peroxide-induced cytotoxicity (73.8%, p < O.O1) was blocked by the extract at concentrations of 50, and 100, 250, 500 ${\mu}$g/ml significantly (p < 0.05, and p < O.O1). The extract at concentrations of 10. and 50 ${\mu}$g/ml decreased the MDA formation on hydrogen peroxide-induced lipid peroxidation. Conclusions : In conclusion, the extract of Panax ginseng has potent antioxidant activity and increases the survival rate of GC-1 spg cells against $H_20_2$-induced cytotoxicity.