• Journal of Acupuncture Research
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• v.29 no.1
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• pp.67-74
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• 2012

Objectives : The objective of this study is to study the effects of hot aqueous extract and ethanol extract from $Lonicera$ $japonica$ $Flos$ on nitric oxide(NO) and prostaglandin $E_2(PGE_2)$ production in macrophage. Methods : $Lonicera$ $japonica$ $Flos$ was extracted in two ways. One was extracted with distilled water(2L) for 4 h and the other one was extracted with 70% ethanol (2L) for 4h. The RAW 264.7 macrophage was subclutured. In order to evaluate cytotoxicity, MTT assay was performed. The concentrations of NO were measured by Griess assay. The concentrations of $PGE_2$ were measured by enzyme immunoassay. Results : 25, $125{\mu}g/m{\ell}$ hot aqueous extract from $Lonicera$ $japonica$ $Flos$ inhibited NO production in LPS-stimulated RAW 264.7 macrophages significantly. 25, 125, $625{\mu}g/m{\ell}$ ethanol extract from $Lonicera$ $japonica$ $Flos$ inhibited NO production in LPS-stimulated RAW 264.7 macrophages significantly. 150, $200{\mu}g/m{\ell}$ hot aqueous extract and ethanol extract from $Lonicera$ $japonica$ $Flos$ inhibited $PGE_2$production in LPS-stimulated RAW 264.7 macrophages significantly. Conclusions : This study suggests that hot aqueous extract and ethanol extract from $Lonicera$ $japonica$ $Flos$ suppress NO and $PGE_2$ production. So hot aqueous extract and ethanol extract from $Lonicera$ $japonica$ $Flos$ may have an anti-inflammation effect.

• Journal of Pharmacopuncture
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• v.17 no.1
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• pp.7-12
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• 2014

Objectives: The objective of this study is to investigate the effects of Salviae Miltiorrhizae Radix hot aqueous extract on nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production and on 1,1-diphenyl-2-picryl hydrazyl (DPPH) free-radical scavenging in macrophages. Methods: Salviae Miltiorrhizae Radix (300 g) was heated at $100^{\circ}C$ with distilled water (2 L) for 4 hours. The extract was filtered and concentrated to 100 mL by using a rotary evaporator, was frozen at $-80^{\circ}C$, and was then freeze-dried by using a freezing-drying system. The RAW 264.7 macrophage was subcultured by using $10-{\mu}g/mL$ lipopolysaccharide (LPS). In order to evaluate cytotoxicity, we performed 3-(4,5-dimrthylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays and measured the cell viability. The NO production was measured by using Griess assays, and the $PGE_2$ production was measured by using enzyme immunoassays. The antioxidant activity, the 1,1-diphenyl-2-picryl hydrazyl (DPPH) free-radical scavenging capability, was measured by using the DPPH method. Results: Cell viability with the 1-, 5-, 25-, 125- and $625-{\mu}g/mL$ Salviae Miltiorrhizae Radix hot aqueous extract was not significantly decreased compared to the cell viability without the extract. When 125 and $625{\mu}g/mL$ of Salviae Miltiorrhizae Radix hot aqueous extract were used, nitric oxide (NO) production in LPS-stimulated RAW 264.7 macrophages was significantly inhibited compared to that in the control group. When 25, 125, and $625{\mu}g/mL$ of Salviae Miltiorrhizae Radix hot aqueous extract were used, $PGE_2$ production in LPS-stimulated RAW 264.7 macrophages was significantly inhibited compared to that in the control group. The 125- and $625-{\mu}g/mL$ Salviae Miltiorrhizae Radix hot aqueous extracts had high DPPH free-radical scavenging capabilities in RAW 264.7 macrophages. Conclusion: This study indicates that Salviae Miltiorrhizae Radix hot aqueous extract suppresses NO and $PGE_2$ production and improves DPPH free-radical scavenging capability. Thus, it seems that Salviae Miltiorrhizae Radix hot aqueous extract may have an anti-inflammation effect and antioxidant activity.

• Journal of Acupuncture Research
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• v.31 no.1
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• pp.131-137
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• 2014

Objectives : This study is to investigate the effects of Acanthopanacis Cortex hot aqueous extract on nitric oxide(NO), prostaglandin E2(PGE2) production and DPPH(1,1-diphenyl-2-picryl hydrazyl) radical scavenging activity in macrophages. Methods : Acanthopanacis Cortex(200 g) was heated at $100^{\circ}C$ with distilled water(2 L) for 4hrs. The extract was filtered and concentrated to 100 ml using a rotary evaporator and was frozen at $-80^{\circ}C$, then was freeze-dried. The RAW 264.7 macrophages were subcultured. In order to evaluate cytotoxicity, MTT assay was performed. Experimental groups were divided into five(control, AC 25, 50, 100 and 200 ${\mu}g/ml$) and we measured cytotoxicity. The concentrations of NO were preprocessed by Griess assay. The RAW 264.7 macrophages was pretreated by 10 ${\mu}g/ml$ LPS and experimental groups were divided into five and we measured NO production. The concentrations of $PGE_2$ were measured by enzyme immunoassay. The RAW 264.7 macrophages was pretreated by 10 ${\mu}g/ml$ LPS. Experimental groups were divided into five and we measured $PGE_2$ production. Antioxidant activity was measured by the DPPH method. experimental groups were divided into four(AC 25, 50, 100 and 200 ${\mu}g/ml$) and we measured DPPH radical scavenging activity. Results : 1. Viability of RAW 264.7 macrophages did not significantly decrease in 25, 50 and 100 ${\mu}g/ml$ Acanthopanacis Cortex hot aqueous extract compared to control group. 2. NO production in LPS-stimulated RAW 264.7 macrophages significantly inhibited in 100, 200 ${\mu}g/ml$ Acanthopanacis Cortex hot aqueous extract compared to control group. 3. $PGE_2$ production in LPS-stimulated RAW 264.7 macrophages significantly inhibited in 100, 200 ${\mu}g/ml$ Acanthopanacis Cortex hot aqueous extract compared to control group. 4. DPPH radical scavenging capability of Acanthopanacis Cortex hot aqueous extract in RAW 264.7 macrophages had the high level in 100, 200 ${\mu}g/ml$. Conclusion : According to the results, Acanthopanacis Cortexx hot aqueous extract has ability to suppress NO, $PGE_2$ production and improve DPPH free radical scavenging activity. So Acanthopanacis Cortex hot aqueous extract may have an anti-inflammation effect and antioxidant activity.

• Journal of Acupuncture Research
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• v.35 no.3
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• pp.115-119
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• 2018

Background: The objective of this study was to determine the effects of Daegangwhal-Tang (DGHT) hot aqueous extract on production of inflammatory mediators and antioxidants in RAW 264.7 macrophage. Methods: DGHT was extracted with water, filtered, concentrated and freeze-dried to perform. Cytotoxicity of DGHT extract was performed by MTT assay. Activated macrophages were treated with varying concentrations of DGHT extract (10, 50, 100 and $200{\mu}g/mL$), and nitric oxide (NO) and prostaglandin E2 ($PGE_2$) concentrations were measured to detect anti-oxidative effects. Interleukin-6 (IL-6), interleukin-1 beta ($IL-1{\beta}$) and tumor necrosis factor-alpha($TNF-{\alpha}$) concentrations were also measured to detect inflammatory responses to DGHT Results: Cytotoxicity of DGHT extract at concentrations of 10, 50, 100 and $200{\mu}g/mL$ were not observed. NO production was significantly decreased in the DGHT hot aqueous extract $200{\mu}g/mL$ concentration group. $PGE_2$, IL-6, $IL-1{\beta}$ and $TNF-{\alpha}$ production was significantly decreased in the DGHT hot aqueous extract 100 and $200{\mu}g/mL$ concentration groups. DGHT hot aqueous extract appeared to have DPPH free radical scavenging capability at all of concentrations, but did not exceed 50%. Conclusion: These results suggest that DGHT hot aqueous extract has concentration-dependent anti-inflammatory and anti-oxidative effect.

• Journal of Acupuncture Research
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• v.28 no.1
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• pp.77-84
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• 2011

Objectives : The objective of this study is to study the effects of hot aqueous extract and ethanol extract from Paeoniae Radix Rubra on nitric oxide(NO) and prostaglandin $E_2(PGE_2)$ production in macrophage. Methods : Paeoniae Radix Rubra were extracted in 2 ways. one was extracted with hot aqueous for 4 hr in $100^{\circ}C$ and the other one was extracted with 70% ethanol for 4 hr in $70^{\circ}C$. RAW264.7 cells, a mouse macrophage lines, were incubated with different concentrations of the extract for 30 min and then stimulated with LPS at indicated times. Cell toxicity was determined by MTT assay. The concentrations of NO and $PGE_2$ were measured by griess assay and enzyme immunoassay(EIA). Results : The hot aqueous and ethanol extracts of Paeoniae Radix Rubra significantly inhibited the NO productions in LPS-stimulated RAW264.7 cells. The hot water extract of Paeoniae Radix Rubra significantly inhibited the $PGE_2$ productions in LPS-stimulated RAW264.7 cells. Conclusions : Our results demonstrated that Paeoniae Radix Rubra extract is able to significantly inhibit the production of NO, $PGE_2$ expression. Hot aqueous extract of Paeoniae Radix Rubra has more effective anti-inflammation than ethanol extract.

• Journal of Acupuncture Research
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• v.34 no.2
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• pp.83-91
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• 2017

Objectives : Alpiniae oxyphyllae Fructus (AOF) is an herbal medicine, which has been used for the treatment of fatigue, chills, and poor physical conditions. The objective of this study was to investigate the anti-inflammatory and anti-oxidative effects of AOF hot aqueous extract. Methods : The cytotoxicity of AOF extract was evaluated using the MTT assay. Nitric oxide (NO) production was measured by the Griess reaction. Prostaglandin $E_2$ ($PGE_2$) production was measured by a commercial competitive enzyme immunoassay. Cytokine production (IL-1tion co6, and TNF- F- was measured by ELISA. The anti-oxidative effect of AOF extracts was measured by the DPPH method. Polyphenol and flavonoid contents were measured by Folin-Ciocalteu's phenol reagent and aluminum chloride, respectively. Results : AOF hot aqueous extract did not show toxicity at doses of 25, 50, 100, and $200{\mu}g/mL$. AOF extract significantly inhibited NO production at doses of 100 and $200{\mu}g/mL.PGE_2$ production was inhibited by AOF extract treatment at doses of 100 and $200{\mu}g/mL$. AOF extracts reduced IL-6 production in a dose-dependent manner. IL-1ent maTNF- F- 1ent mannerd IL-6 production in uction at doses of 100 and ${\mu}g/mL$. The DPPH free radical scavenging capability was above 50% at $200{\mu}g/mL$. Conclusion : This study suggests that AOF hot aqueous extract may exert anti-inflammatory and anti-oxidative effects in a dose-dependent manner. Further studies are required for validating the safety and efficacy of AOF.

• Journal of Acupuncture Research
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• v.35 no.1
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• pp.41-45
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• 2018

Background: Danggwisusan is a herbal medicine which is used to treat bruises, static blood, external injuries, and somatalgia in Korean medicine. The objectives of this study were to investigate whether Danggwisusan hot aqueous extract had an inhibitory effect upon inflammatory cytokine production and oxidation. Methods: Cytotoxic activity of Danggwisusan extract was examined by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. The amount of nitric oxide produced was measured using Griess reagent. Prostaglandin E2 production was measured using an enzyme immunoassay. Inflammatory cytokines ($IL-1{\beta}$, IL-6 and $TNF-{\alpha}$) were measured by an enzyme linked immunosorbent assay. The anti-oxidative effect of Danggwisusan was measured by the 1,1-Diphenyl-2-picryl hydrazyl method. The amount of polyphenol and flavonoid contents were measured by Folin and Ciocalteauea phenol reagent and aluminum nitrate. Results: Danggwisusan hot aqueous extracts did not show significant toxicity at 10, 20, 50, and $100{\mu}g/mL$. At a dose of $100{\mu}g/mL$, Danggwisusan hot aqueous extract significantly inhibited nitric oxide and $PGE_2$ production, and significantly reduced $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production. At a dose of $100{\mu}g/mL$, 1,1-Diphenyl-2-picryl hydrazyl free radical scavenging capability was over 50%. Conclusion: This study showed that Danggwisusan hot aqueous extract may have anti-inflammatory and anti-oxidative effects on macrophages.

• Advances in Traditional Medicine
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• v.2 no.2
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• pp.101-105
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• 2002

A battery of neuropharmacological experiments showed the hot water extract of Ocimum sanctum Linn. (Labiateae) had a depressant effect on the central nervous system (CNS), but the aqueous extract showed no effect on it. The hot water extract reduced the spontaneous locomotor activity, exploratory head dipping, propulsive locomotion and exploratory ambulation as well as prolonged the pentobarbital induced sleeping time. The depressant effect starts from 60 minutes after the drug administration and continued to 180 minutes. The drug may exert central depressant effect by interfering with the function of the cortex.

• Journal of Marine Bioscience and Biotechnology
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• v.10 no.2
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• pp.53-61
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• 2018

Sargassum fulvellum (gulfweed) is a widespread seaweed in the coastal areas of northeast Asia. In the present study, we identified the phenolic compounds present in aqueous and ethanolic extracts of S. fulvellum and evaluated their antioxidative properties and their abilities to block cell proliferation using in vitro assays: antioxidant activity was assessed by using a DPPH assay and superoxide anion scavenging activity, anti-tyrosinase activity, and anti-proliferative activity were assessed using MTT and lactate dehydrogenase [LDH] assays in vascular smooth muscle cells. The hot-water ($65^{\circ}C$) extract had a higher phenol content than the ethanolic extract. The hot-water extract showed a statistically significant increase in free radical scavenging activity and a greater ability to reduce proliferation of vascular smooth muscle cells stimulated with platelet-derived growth factor-BB. Taken together, hot-water extracts of S. fulvellum may be an important source of antioxidative and antiproliferative agents.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.2
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• pp.255-261
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• 2004

Antioxidant activity of extracts from the submerged-liquid culture of mushrooms was measured using two systems : linoleic acid and mouse liver microsomes induced by various free radical sources. Mushrooms of Pleurotus ostreatus (Neutari), Phellinus linteus (Sanghwang), Paecilomyces japonicus (Dongchunghacho), Hericicum erinacium (Norugungdengyee) and Agaricus blazei (Shinryeong) in 1% mulberry tree powder-supplemented medium were incubated in a shaking incubator (200 rpm, $25^{\circ}C$) for 3 days. Hot water extracts of mycelial cultures were freeze-dried, followed by fractioning with hexane, chloroform, ethylacetate and butanol in the order. Antioxidant activity of each sample was examined in free radical-induced linoleic acid oxidation in phosphate-buffered saline (PBS ) solution by measuring the amount of malonaldehyde (MA), and mouse liver microsomal systems by measuring the amount of thiobarbituric acid reactive substances (TBARS). In linoleic acid oxidation system, hot water extracts from the cultures of Pleurotus ostreatus, Phellinus linteus, and Paecilomyces japonicus exhibited stronger antioxidant activity than aqueous or butanol fraction and the combined fraction of hexane, chloroform and ethylacetate, but the hot water extract from Pleurotus ostreatus culture was the strongest activity. The antioxidant activity of the hot water extract from Pleurotus ostreatus culture was stronger than any other fractions in mouse microsomal system. These results suggest that hot water extract of Pleurotus ostreatus culture, and the cultures of Phellinus linteus and Paecilomyces japonicus could be useful for functional materials to reduce the oxidation of lipids in food systems induced by free radicals.