• The Journal of Pediatrics of Korean Medicine
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• v.30 no.3
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• pp.1-30
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• 2016

Objectives Previous studies have found out that Forsythiae Fructus (FF) extracts have anti-atopic activities by in vitro experiment. In order to understand more about FF extracts' benefit, we subdivided FF extracts depending on systematic fractionation method by using Methylene chloride (MC), Ethyl acetate (EtOAc), n-BuOH and n-hexane (n-Hx). This study is designed to examine the effect of FF fractions on the PMA- ionomycin-induced activation of RBL-2H3 mast cell lines in vitro and on the DNCB-induced activation of NC/Nga mice in vivo. Methods For this study, we examined IL-4, IL-13 production by ELISA analysis, IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$ mRNA expression by real-time PCR and manifestations of AP-1 and MAPKs transcription factors by western blotting in vitro. Through in vitro experiment, we selected FF n-BuOH fraction that seems the best effective in atopic dermatitis then induced it on NC/Nga mice by DNCB. We measured mice's WBC, eosinophil and neutrophil in heart blood, IL-4, IL-5, IFN-${\gamma}$ in the spleenocyte culture supernatant, the absolute cell numbers of CD4+, CD8+, B220+CD23+, CD3+CD69+ and Gr-1+CD11b+ in the PBMCs, ALN and dorsal skin, IL-5, IL-13, IL-31, IL-31RA in the dorsal skin by real-time PCR and the distribution of immune cells by H&E on dorsal skin and ANL and toluidine blue staining on dorsal skin. Results FF n-BuOH fraction suppressed IL-4, IL-13 production and mRNA expression of IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$. Results from the western blot analysis showed that FF n-BuOH fraction reduced the activation of the mast cell specific transduction factors involved in AP-1 by suppressing JNK and ERK phosphorylation. In the gross, atopic dermatitis induced by DNCB in NC/Nga mice were improved by oral administration of FF n-BuOH fraction. Oral FF n-BuOH fraction also decreased the level of IgE in mice's serum and the level of IL-4 and IL-5 in the spleenocyte culture supernatant, cell numbers of CD8+, B220+CD23+ in the PBMCs, CD4+ in the ALN and CD4+, Gr-1+CD11b+ in the dorsal skin and suppressed mRNA expression of IL-5, IL-13, IL-31, IL-31RA in the dorsal skin. Histological examination showed that infiltration levels of immune cells in atopic dermatitis induced NC/Nga mice were improved by FF n-BuOH fraction. Conclusions FF n-BuOH fraction can reduce pruritus by suppressing IL-31, IL-31RA secretion and modulate molecular mediators and immune cells associated with atopic dermatitis induced in NC/Nga mice which may have played a significant role in recovering atopic dermatitis symptoms.

• The Korea Journal of Herbology
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• v.27 no.3
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• pp.23-29
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• 2012

Objective : Sanguisorbae Radix(SO). that belong to Rosaceae is widely distributed in Asia including Korea, Japan and China. It has been used as traditional medicine from old times as a treatment for anti-inflammatory drugs. This study was designed to investigate the effects of n-BuOH fractions of SO on anti-oxidant effect and anti-microbial effect on $Streptococcus$ $mutans$ ($S.$ $mutans$). Methods : The anti-microbial effect of $n$-BuOH fractions of SO. was assessed by the paper disk diffusion method and anti-oxidant effect was assessed by the DPPH radical scavenging effect, Superoxide anion radical scavenging effect and SOD like ability. Results : DPPH radical scavenging of the $n$-BuOH fraction from SO in 50 ug/mL was shown to be Fr-2 (89.9%), Fr-3 (88.7%), Fr-4 (76.3%), Fr-1 (59.4%), Fr-5 (56.2%). Superoxside anion radical scavenging activity of the n-BuOH fraction from SO. in 50 ug/mL for Fr-3 was shown to be 78%. SOD-like activity of n-BuOH fraction from the SO in 1000 ug/mL for Fr-3 were shown to be 76.1% respectively. The $n$-BuOH fraction from the SO had high anti-microbial effect on $S.$ $mutans$. Conclusion : As a result, the $n$-BuOH fraction from SO. has good anti-microbial and anti-oxidant effects in a concentration-dependent manner.

• Journal of Life Science
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• v.26 no.8
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• pp.948-954
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• 2016

We investigated the flavonoid and phenol contents and antioxidant effect of wine by-product extract. Antioxidant effects were measured with 1,1-diphenyl-2-picryhydrazyl (DPPH) and 2.2'-azino-bis(3-ethylbenothiazoline-6-sulfonic acid) diammonium salt radical cation (ABTS+) assays. Cellular reactive oxygen species (ROS) were measured with the dichlorofluorescein-diacetate (DCFH-DA) assay. The flavonoid and phenol contents of the methanol (MeOH) extract were greater than those of the acetone+methylene chloride (A+M) extract. Among fractions, the 85% aqueous methanol (85% aq. MeOH) fraction contained the highest flavonoid contents, while the n-BuOH fraction had more phenol contents. In the DPPH and ABTS assays, the MeOH extract showed a scavenging effect greater than that of the A+M extract (p<0.05). The n-BuOH fraction (0.5 mg/ml concentrations) showed scavenging effects of 72% and 92%, respectively, in the DPPH and ABTS assays (p<0.05). However, the 85% aq. MeOH fraction showed a 90% scavenging effect in the DPPH assay only. In 120 min ROS production assay, all tested fractions dose-dependently decreased cellular ROS production induced by H₂O₂ in comparison with that produced by exposure to the extract-free control. The MeOH extract showed a higher sinhibitory effect on cellular ROS producing than that of the A+M extract at all concentrations tested. Treatment with the n-BuOH fraction (0.1 mg/ml concentrations) inhibited cellular ROS production by 60%. These results indicate that the n-BuOH fraction of wine by-product extract inhibited cellular oxidation and may contain valuable bioactive compounds such as flavonoids and phenols.

• The Journal of Korean Medicine
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• v.23 no.4
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• pp.45-54
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• 2002

Objectives: In order to investigate the relationship of Radix Trichosanthis components and the melanin synthesis, the author has analyzed the cell viability and tyrosinase activity, melanin content and morphologic changes in n-hexane, EtOAc, n-BuOH, and H2O fraction. Methods: At first, in order to determine the concentration of the Radix Trichosanthis component, the author investigated the viability of B16 melanoma cell. To measure the effects of Trichosanthes kirilowii extracts (n-BuOH, n-Hexane, EtOAc, H2O fractions) on the viability of A549 cells, A549 cells were treated with various concentrations (from 0.5 to $25{\;}\mu\textrm{g}/ml$) of components of Trichosanthes kirilowii. After 24hrs, the cell viability was measured by MTT assay. The EtOAc components of Trichosanthes kirilowii decreased the viability of A549 cells in a dose-dependent manner. H2O and n-BuOH components had no cell toxicity till $25{\;}\mu\textrm{g}/ml$, the n-hexane component showed minor cell toxicity at $25{\;}\mu\textrm{g}/ml$ and the EtOAc component cell toxicity was revealed at $5{\;}\mu\textrm{g}/ml$ concentration. Results: 1. The results of tyrosinase activity and the Radix Trichosanthis component; n-hexane and EtOAc components controlled it effectively; the n-BuOH components were less effective. 2. The results of melanin content analysis showed that the n-hexane and EtOAc components effectively inhibited, the n-BuOH fraction inhibited less, and H2O component didn't inhibit the terminal melanin formation. 3. In the n-BuOH and H2O component there were no changes, but in the n-hexane component the melanin content was effectively inhibited. 4. In the EtOAc fraction, although the melanin content was inhibited, the cell count was evidently suppressed, Of all of the Radix Trichosanthis components, the n-Hexane and EtOAc fractions inhibited the melanin synthesis best, but owing to its toxicity, the EtOAc components inhibited the cell count. Conclusion: The above results demonstrated that Radix Trichosanthis n-hexane fraction efficiently inhibited the tyrosinase activity and melanin synthesis.

• Journal of Life Science
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• v.15 no.1 s.68
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• pp.106-111
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• 2005

To investigate the antioxidant activity of extract from the raw walnut, Juglans sinensis Dode, we prepared five fractions (methanol (MeOH), dichloromethane $(CH_2Cl_2)$, ethyl acetate (EtOAc), n-buthanol (n-BuOH) and dehydrogen monooxide $(H_2O)$ fractions) and examined. The effect of walnut extract on the oxidative stress was investigated in vitro. The DPPH (2,2-Di (4-tert-octylphenyl)-1-picrylhydrazyl) free radical scavenging activity of extract from raw walnut was shown in the following order: $EtOAc\;fraction layer. The result showed that the highest activity $(0.56{\mu}g/ml,\;IC_{50}.)$ was observed in EtOAc fraction, whereas n-BuOH fraction, MeOH fraction, $CH_2O_2$ fraction and $H_2O$ layer of $IC_{50}$ were $2.34{\mu}g//ml,\;3.88{\mu}g/ml,\;8.06{\mu}g/ml,\;and\;8.19{\mu}g/ml$, respectively. The radical scavenging activity assay of each fraction showed that the antioxidative activity was observed in the following order: EtOAc fraction $(74.27\pm1.56\%)>MeOH\;fraction\;(60.76\pm3.4\%)>n-BuOH\;fraction\;(59.32\pm0.88\%)>H_2O\;layer\;(41.69\pm2.06\%)$. These results revealed that all fractions, except for $CH_2Cl_2$ fraction, showed high antioxidative activity. Furthermore, the peroxynitrite $(ONOO^-)$ scavenging activity was assayed in each fraction. The result showed that the $ONOO^-$ scavenging activity of EtOAc fraction, MeOH fraction and n-BuOH fraction from raw walnut was $95.14\pm0.36\%,\; 90.02\pm1.19\%\;and\;89.41\pm0.81\%$, respectively. The tert-butylhydroperoxide (t-BHP) treatment in vitro increased lactate dehydrogenase release and lipid peroxidation in renal cortical slices. Such changes were completely prevented by addition of MeOH fraction, EtOAc fraction and n-BuOH fraction of walnut. These results indicate that the walnut extract exerts the benedicial effect against t-BHP-induced cell injury and its effect may be due to antioxidant action. In addition, it is suggested that walnut extract might be developed as the effective scavenger for the prevention of oxidative stress.

• Journal of Life Science
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• v.23 no.3
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• pp.361-367
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• 2013

Hizikia fusiformis has been widely used in Oriental herbal medicine and health food. To identify antioxidation properties that contain natural bioactive substances, we investigated the distribution of active compounds existing in batches of organic solvent fractionation. A dried form of H. fusiformis was subjected to sequential fractionation using n-hexane, ethyl acetate, n-BuOH, and aqueous n-BuOH. The results showed that among the four isolated fractions, the n-BuOH fraction showed the highest antioxidation activities. The n-BuOH fraction was applied to reserve-phase silica gel column chromatography, which produced three fractions: BA, BB, and BC. Among these fractions, BB showed the highest antioxidation activities, which increased in a concentration-dependent manner. At a concentration of 1.0 mg/ml n-BuOH fraction, the activities of DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging and reducing power were approximately $45{\pm}0.14%$ and $1.34{\pm}0.23$, respectively. In addition, the activities of ${\beta}$-carotene-linoleic acid, hydrogen peroxide scavenging, and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)) radical scavenging were $76{\pm}0.12%$, $82{\pm}0.06%$, and $65{\pm}0.17%$, respectively. These findings suggest that the BB fraction contains potent antioxidation properties and that it could be used in the production of natural and functional foods.

• KSBB Journal
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• v.19 no.1
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• pp.67-71
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• 2004

An antioxidant activity of Rosa rugosa extract and its solvent-partitioned fractions was determined not only by measuring lipid peroxide produced when a mouse liver homogenate was exposed to the air at 37$^{\circ}C$, using thiobarbituric acid (TBA) but also by evaluating the free radical scavenging effect against DPPH radical, authentic peroxynitrite, and 3-morpholinsydnonimine (SIN-1). All its partitioned fractions including crude extract showed potent scavenging effect against DPPH radical, peroxynitrite, and lipid peroxidation. n-BuOH fraction, in particular, was found to be the most effective in DPPH radical scavenging ability as well as inhibition against lipid peroxidation. The 15% aqueous MeOH fraction also showed a strong potency which was slightly lower than n-BuOH fraction. Based on these results, we suggest that Rosa rugosa could be useful for preventing an oxidative damage.

• Journal of Life Science
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• v.29 no.3
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• pp.337-344
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• 2019

The free radical scavenging, cytotoxic effects, and flavonoid content of fractions from Lycopus lucidus Turcz leaves were here investigated. The flavonoid contents of 85% methanol (MeOH) and n-butanol (BuOH) fractions of the leaves were 41.5 mg/g and 77.2 mg/g, respectively. In DPPH and ABTs+ assays, 85% MeOH and n-BuOH fractions from the L. lucidus Turcz leaves had a greater scavenging effect (p<0.05). The n-BuOH fraction (0.5 mg/ml concentration) had scavenging effects of 88% and 92% in the DPPH and ABTs+ assays, respectively (p<0.05). Cell viability tests showed that treatment with L. lucidus Turcz leaf fractions caused cytotoxicity in the growth of AGS, HT-29, and HT-1080 cancer cells. Of the different fractions, the 85% MeOH sample displayed the highest cytotoxic activity; the $IC_{50}$ values of this fraction against AGS, HT-1080, and HT-29 cancer cells were 0.03 mg/ml, 0.14 mg/ml, and 0.16 mg/ml, respectively. These biological results indicate that the n-BuOH fraction was more effective in anti-oxidant activity while the 85% MeOH fraction was stronger in cytotoxic effects, and they suggest that these two fractions from L. lucidus Turcz leaves may contain valuable bioactive compounds, such as flavonoids.

• International Journal of Industrial Entomology and Biomaterials
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• v.17 no.2
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• pp.197-200
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• 2008

The antioxidant activity of Isaria (Paecilomyces) sinclairii was determined by measuring its radical scavenging effect on 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radicals. The n- BuOH extract of P. sinclairii showed strong scavenging activity to DPPH. The anti-oxidant potential of the individual fraction was in the order of ethylacetate> n- BuOH>chloroform>n-hexane. The n-BuOH soluble fraction exhibiting strong anti-oxidant activity was further purified by repeated silica gel column chromatography. N-(2-Hydroxyethyl)adenosine (HEA) was isolated as one of the active principles from the n-BuOH layer. The n-BuOH layer, particularly HEA, did not increase the level of nitric oxide (NO) production in vascular endothelial cells that might be related to vasorelaxation such as the action of viagra. In addition, the vascular endothelial growth factor (VEGF) levels showed little or no increase compared with control group with the treatment of I. sinclairii.

• Food Science and Preservation
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• v.18 no.1
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• pp.65-71
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• 2011

We explored the effect of extracts of dried Platycodon grandiflorum on production of reactive oxygen species (ROS), glutathione (GSH) and nitric oxide (NO). To determine antioxidant activity in the presence of $H_2O_2$-induced oxidative stress, DCFH-DA (dichlorodihydrofluorescin diacetate) assay was employed. Acetone/methylene chloride (A+M) and methanolic (MeOH) extracts of P. grandiflorum reduced intracellular ROS levels. Of the various tested fractions, n-BuOH fraction showed the highest protective effect in terms of lipid peroxide production. Total GSH levels were measured after treatment of HT1080 cells with the A+M and MeOH extracts, and other solvent fractions, at various concentration. The A+M extacts and 85% (v/v) aqueous MeOH fraction significantly increased GSH levels (p<0.05). When lipopolysaccharide (LPS)-induced NO production was evaluated, all tested crude extracts, and fractions thereof, significantly reduced NO production (p<0.05), and the n-BuOH and 85% (v/v) aqueous MeOH fractions (at 0.05 mg/mL) showed the strongest inhibitory effects. The results showed that the n-BuOH fraction inhibited both cellular oxidation and NO production, and this fraction may thus contain valuable active compounds.