Objectives : Woogakseungmatang is a prescription medication mainly used to treat facial paralysis in Korean medicine. The purpose of this study is to investigate the effects of Woogakseungmatang on anti-inflammation and anti-oxidation. Methods : Woogakseungmatang was extracted using hot water. Cytotoxicity was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) method; nitric oxide(NO) production and Prostaglandin $E_2$ ($PGE_2$) production in RAW cells treated with Woogakseungmatang were investigated; and the cytokine changes associated with inflammation were examined. The antioxidant capacity of Woogakseungmatang was measured using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. Results : RAW cells treated with Woogakseungmatang showed 90% cell viability at a $100-{\mu}g/ml$ concentration. NO production was decreased by 15% at a $100-{\mu}g/ml$ concentration. $PGE_2$ production was decreased by 18% at a $100-{\mu}g/ml$ concentration. Interleukin $1{\beta}$ ($IL-1{\beta}$), interleukin 6(IL-6), and tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$) were significantly reduced at $100{\mu}g/ml$ compared with those in the control group. The DPPH free radical scavenging capability was more than 50% at $100{\mu}g/ml$. Conclusions : Woogakseungmatang showed only a slight anti - inflammatory effect at $100{\mu}g/ml$ and it was difficult to confirm the concentration-dependent anti-inflammatory effect. Therefore, this study means to confirm the potential anti-inflammatory effects of Woogakseungmatang. Based on this research, more systematic and diverse studies should be conducted.
Hossen, Muhammad Jahangir;Hong, Yong Deog;Baek, Kwang-Soo;Yoo, Sulgi;Hong, Yo Han;Kim, Ji Hye;Lee, Jeong-Oog;Kim, Donghyun;Park, Junseong;Cho, Jae Youl
Journal of Ginseng Research
/
v.41
no.1
/
pp.43-51
/
2017
Background: BIOGF1K, a compound K-rich fraction prepared from the root of Panax ginseng, is widely used for cosmetic purposes in Korea. We investigated the functional mechanisms of the anti-inflammatory and antioxidative activities of BIOGF1K by discovering target enzymes through various molecular studies. Methods: We explored the inhibitory mechanisms of BIOGF1K using lipopolysaccharide-mediated inflammatory responses, reporter gene assays involving overexpression of toll-like receptor adaptor molecules, and immunoblotting analysis. We used the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay to measure the antioxidative activity. We cotransfected adaptor molecules, including the myeloid differentiation primary response gene 88 (MyD88) and Toll/interleukin-receptor domain containing adaptor molecule-inducing interferon-${\beta}$ (TRIF), to measure the activation of nuclear factor (NF)-${\kappa}B$ and interferon regulatory factor 3 (IRF3). Results: BIOGF1K suppressed lipopolysaccharide-triggered NO release in macrophages as well as DPPH-induced electron-donating activity. It also blocked lipopolysaccharide-induced mRNA levels of interferon-${\beta}$ and inducible nitric oxide synthase. Moreover, BIOGF1K diminished the translocation and activation of IRF3 and NF-${\kappa}B$ (p50 and p65). This extract inhibited the upregulation of NF-${\kappa}B$-linked luciferase activity provoked by phorbal-12-myristate-13 acetate as well as MyD88, TRIF, and inhibitor of ${\kappa}B$ ($I{\kappa}B{\alpha}$) kinase ($IKK{\beta}$), and IRF3-mediated luciferase activity induced by TRIF and TANK-binding kinase 1 (TBK1). Finally, BIOGF1K downregulated the NF-${\kappa}B$ pathway by blocking $IKK{\beta}$ and the IRF3 pathway by inhibiting TBK1, according to reporter gene assays, immunoblotting analysis, and an AKT/$IKK{\beta}$/TBK1 overexpression strategy. Conclusion: Overall, our data suggest that the suppression of $IKK{\beta}$ and TBK1, which mediate transcriptional regulation of NF-${\kappa}B$ and IRF3, respectively, may contribute to the broad-spectrum inhibitory activity of BIOGF1K.
G. japonicum is a perennial hem and the flowering plant has been used as a diuretic and an astringent in Japan and China. However, little information is available about the anti-inflammatory action of G. japonicum. Therefore, the objective of this study was to investigate the antiinflammatory action of fractions from G. japonicum methanol extract. Inhibition of NO production was observed when cells were cotreated with fractions of G. japonicum and lipopolysaccharide. We observed that ethyl acetate fraction of G. japonicum inhibited NO production by LPS-activated RAW 264.7 cells, and that the suppression induced by ethyl acetate fraction of G. japonicum was associated with antioxidant activity and direct NO clearance. In addition, only ethyl acetate fraction of G. japonicum inhibited stimulated $PGE_2,\;TNF-\alpha,\;IL-1\beta$ production, whereas water and methyl chloride fractions showed no such effects. The ethyl acetate fraction of G. japonicum methanol extract showed a remarkable scavenging activity on the 1,1-diphenyl-2 picrylhydrazyl radical. Based on the results, ethyl acetate fraction of G. japonicum may be useful source as natural antioxidants and antiinflammation. Therefore, the results obtained from this study provide an alternative protective mechanism of ethyl acetate fraction of G. japonicum and provide information on the potential use of ethyl acetate fraction of G. japonicum in chemoprevention or pathogenic conditions related to overproduction of NO and $PGE_2$. However, the mechanism of the inflammatory effect must be evaluated through various parameters for induction of NO production.
The mulberry tree (Morus alba L.) has been traditionally used in Chinese medicine to treat inflammatory diseases. We investigated the effects of bioconversion on different components of the mulberry tree, and determined changes in the physiological activities. Ethyl acetate-soluble fractions of five different segments (fruit, Mori Fructus; leaf, Mori Folium; twig, Mori Ramulus; root, Mori Cortex; and mistletoe, Loranthi Ramulus) of the mulberry tree show enhanced anti-oxidant effects in the 2,2-diphenyl-1-picrylhydrazyl, and 2,2'-azinobis-(3-ethylvenzothiazoline-6-sulfonic acid) assays, and enhanced anti-inflammatory effects of lipopolysaccharide (LPS)-stimulated nitric oxide (NO) production in RAW 264.7 macrophages, after being treated with a crude enzyme extract from Aspergillus kawachii, in the following order of activity: Mori Folium>Mori Cortex>Mori Ramulus>Mori Fructus>Loranthi Ramulus. Ethyl acetate- soluble fraction of mulberry leaves (Mori Folium) that underwent bioconversion was most effective, and was devoid of any cytotoxicity. The fraction was also effective against mRNA expression of LPS-induced pro-inflammatory cytokines, such as inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis $factor-{\alpha}$, $interleukin-1{\beta}$, and interleukin-6. In addition, the fraction was effective in LPS-induced phosphorylation of mitogen-activated protein kinases and IKK, and $I{\kappa}B$ degradation, followed by translocation of the nuclear $factor-{\kappa}B$ from the cytoplasm to the nucleus. Thus, bioconversion increased the anti-oxidative and anti-inflammatory activities of the mulberry leaf.
Kim, Min-A;Lee, Han-Saem;Chon, So-Hyun;Park, Jeong-Eun;Lim, Yu-Mi;Kim, Eun-Jeong;Son, Eun-Kyung;Kim, Sang-Jun;So, Jai-Hyun
Journal of Applied Biological Chemistry
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v.62
no.1
/
pp.39-50
/
2019
Mitogen-activated protein (MAP) kinases play an important role in cell growth and differentiation, as well as the modulation of proinflammatory cytokines. The objective of this study was to examine the increase in the anti-inflammatory effect of Gentiana scabra Bunge (GSB), due to bioconversion with the Aspergillus kawachii crude enzyme, via inhibition of the $NF-{\kappa}B$ signaling and MAP kinase pathways in RAW 264.7 cells. The expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 in RAW 264.7 cells treated with the GSB ethyl acetate fraction bioconverted with A. kawachii crude enzyme (GE-BA), was dramatically suppressed as compared to GSB ethyl acetate fraction non-bioconverted with the A. kawachii crude enzyme (GE-UA). The phosphorylation of p38, extracellular signal-regulated kinases, and inhibitory ${\kappa}B$ in RAW 264.7 cells treated with GE-BA was further suppressed, as compared to exposure to GE-UA. Moreover, the mRNA expression of interleukin 6, interleukin 1-beta, and tumor necrosis $factor-{\alpha}$ was further suppressed by GE-BA, compared to GE-UA. Similarly, anti-oxidant activities, such as 2,2-diphenyl-1-picrylhydrazyl hydrate and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activity, of GE-BA were further increased compared to GE-UA. These observations demonstrate that the anti-oxidant and anti-inflammatory activities of GSB ethyl acetate fraction increases as a result from bioconversion with the A. kawachii crude enzyme.
Yun-Gwon Seon;Jae Min Jeong;Jin-Sol Yoon;Joonyong Noh;Seung Kyu Im;Sung-Pil Bang;Jeong Cheol Shin;Jae-Hong Kim
Journal of Acupuncture Research
/
v.40
no.4
/
pp.356-367
/
2023
Background: The aim of this study is to determine the antioxidant and anti-inflammatory effects of Dusokohwaeum (DOE). Methods: To measure the antioxidant and anti-inflammatory effects of DOE, the total flavonoid and polyphenol contents and radical scavenging activity were measured. Furthermore, reactive oxygen species (ROS), nitric oxide, and cytokine production were measured by treating lipopolysaccharide-induced RAW264.7 cells with DOE, and gene expression levels of inducible cyclooxygenase-2, nitric oxide synthase, and cytokines were evaluated. Results: Radical scavenging experiments revealed a significant concentration-dependent increase in scavenging capacity. The production of ROS, nitric oxide, and cytokines in the cells showed a significant concentration-dependent decrease when compared with the control group. The gene expression levels of inducible cyclooxygenase-2, nitric oxide synthase, and cytokines also showed a significant concentration-dependent decrease when compared with the control group. Conclusion: Interestingly, the antioxidant and anti-inflammatory effects of DOE were 23.42 ± 0.64 mg GAE/g and 20.83 ± 0.98 mg QE/g, respectively. The administration of DOE resulted in a concentration-dependent increase in scavenging ability in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging ability experiments. The production of intracellular ROS and nitric oxide was significantly reduced in the presence of DOE. The production of inflammatory cytokines (prostaglandin E2, tumor necrosis factor-alpha [TNF-α], interleukin-1 beta [IL-1β], and IL-6) was significantly reduced in the presence of DOE. Finally, the expression levels of inducible nitric oxide synthase, cyclooxygenase-2, TNF-α, IL-1β, and IL-6 were significantly decreased in the presence of DOE.
Lee Chi-Hyeoung;Cha Jae-Young;Jun Bang-Sil;Lee Ho-Jun;Lee Young-Chun;Cho Yong-Lark;Cho Young-Su
Journal of Life Science
/
v.15
no.5
s.72
/
pp.819-825
/
2005
The Antioxidative accvities of the cell free extracts containing high glutathione by Saccharomyces cerevisiae FF-8 were tested in vitro experimental models : DPPH method for radical scavenging activity, ferric TBA method and ferric thiocyanate method using linoleic acid and tissue microsome for lipid peroxidation inhibitions. The concentration of intercellular glutathione by cultivating S. cerevisiae FF-8 in the YM optimal medium obtained $204\mug/ml$, which was increased by 2.76-fold from $74\mug/ml$ in the YM basal medium. A comparition between the YM basal medium and the YM optimal medium on antioxidative substance produced by S. cerevisiae FF-8 was investigated. In DPPH ($\alpha, \alpha-diphenyl-\beta-picrylhydrazyl$) method, the electron donating activity of the glutathione produced by S. cerevisiae FF-8 cultured in the YM optimal medium was as high as that of BHT ($ 0.05\%w/v $). The antioxidative a.tivity was measured by inhibition against lipid peroxidation of rat tissues' microsomes. The results of anti-oxidant activity of the cell free extracts by S. rerevisiae FF-8 cultured in the YM optimal medium was shown in the following order . $ liver 60.98\% > kidney 56.43\% > heart 52.91\% > brain 52.13\% > testis 45.57\% > spleen 42.95\% $. In antioxidative activities determined by ferric thiocyanate method and TBA methods against lipid peroxidation, the lipid peroxidation in the control mixture increased more rapidly than the typical peroxidation curve of linoleic acid from one day. The antioxidative activity of the cell free extracts by cultivating S. cerevisine FF-8 in the YM optimal medium were higher than that of the YM basal medium. These data indicate that the cell free extracts containing a high intercellular glutathione of S. cerevisiae FF-8 cultured in YM optimal medium showed strong antioxidative capacities by DPPH radical scavenging activity and ferric thiocyanate and TBARS measurements.
Journal of the Korean Applied Science and Technology
/
v.35
no.4
/
pp.1003-1012
/
2018
This study is for checking the possibility of Lentinula edodes as cosmetic materials. For this we carried out biological active evaluation about anti-oxidant and anti-inflammatory effects by Lentinula edodes extracts. We extracted Lentinula edodes with water and 70% ethanol and then in order to evaluate anti-oxidant activity we treated samples by concentrations (100, 500, 1000) ${\mu}g/ml$ and carried out 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, the activity of 2,2'-azino-bis ( 3-ethylbenzothiazoline-6-sulphonic acid )-diammonium salt (ABTS) cation radical scavenging and superoxide dismutase(SOD) like activity. Also, in order to evaluate effect of anti-inflammatory the samples in macrophages(RAW 264.7 cells), we carried out evaluation of cell viability, nitric oxide inhibitory activity western blot. The results of DPPH, $ABTS^+$ radical scavenging activity and SOD-like activity of the Lentinula edodes extracts increased in dose-dependent manner. The cytotoxic of samples by MTT assay showed no toxicity at the concentrations of 10, 25 and $50{\mu}g/ml$ of Lentinula edodes extract. Nitric oxide inhibition activity results showed that the extracts reduced NO productions in a concentration-dependent manner. Expression of inflammatory cytokines as $TNF-{\alpha}$, $PGE_2$ and $IL-1{\beta}$ decreased in a concentration-dependent manner and iNOS and COX-2 proteins expression rates were decreased significantly in western blot analysis. From the results of the experiment, it was comfirmed that the Lentinula edodes extracts had excellent anti-oxidant and anti-inflammatory effect and could be used as a safe natural cosmetic material in the future.
This study was performed to develop natural spices and functional foods using Cheongung (Cnidium officinale) which is one of the Korean medicinal plants. The volatile flavor patterns of Cnidium officinale were detected by electronic nose with 6 metal oxide sensors, and the principal component analysis was carried out. The volatile flavor components of Cnidium officinale were isolated by simultaneous steam-distillation extraction with pentane and diethylether (1 : 1), and essential oils were analyzed by capillary GC and GC/MS. The free radical scavenging activity of ethanol and methanol extracts from Cnidium officinale was measured by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and compared with ${\alpha}-tocopherol$ as reference. The principal component analysis showed the difference of principal components between fresh and drying samples. Eighty-five volatile flavor components (643.64 ppm) from fresh Cnidium officinale were identified and the major components were butyl phthalide, sabinene, neocnidilide. Sixty-four volatile flavor components (218.15 ppm) from hot air dried one were identified and the major components were butyl phthalide, sabinene, 3-N-butyl phthalide. And 73 volatile flavor components (784.15 ppm) from freeze dried one were identified and the major components were butyl phthalide, sabinene, ${\beta}-selinene$. The free radical scavenging activity of methanol cold extract (500 ppm) of freeze dried Cnidium officinale was higher than other samples. And methanol and ethanol cold extracts (above 250 ppm) of freeze dried sample were higher than ${\alpha}-tocopherol$$25\;{\mu}M$ (22.34%).
For selecting a new candidate as functional material, this study was conducted on in vitro antioxidant activity and total phenol content of methanol and water extracts prepared from two Mentha species (M viridis L. (M spicata L.) and Mentha piperita L, Extracts of M. viridis showed more efficient scavenging activity on superoxide and DPPH ${({\alpha},{\alpha}-diphenyl-{\beta}-picrylhydrazyl)}$ radical and inhibitory activity on oxidation of human low density lipoprotein (LDL) induced by $CuSO_4$ and auto-oxidation of linoleic acid than those of M piprita. Methanol extract $(65.88%{\sim}77.59%)$ and water extract $(37.69%{\sim}87.21%)$ of M. viridis also exhibited more potent inhibitory activity on LDL oxidation than that of ${{\alpha}-tocopherol\;(28.37%{\sim}66.54%)}$ at ${1{\sim}100\;{\mu}g/ml}$ of final concentration. The total phenol contents of methanol extract and water extract of Mviridis (17.95% & 10.18%, respectively) as tannic acid equivalent were higher than those of M piperita (15.44% & 9.19%). But the yields of methanol and water extracts of M. viridis (13.3 % & 13.5%) were lower than those of M. piperita (14.1 % & 14.6%). The results implies that the extracts from M. viridis (spicata) is more useful material for industrialization as functional food than those from M. piperita.
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