• Journal of Animal Reproduction and Biotechnology
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• v.37 no.3
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• pp.183-192
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• 2022

Lipopolysaccharide (LPS) is an endotoxin factor present in the cell wall of Gram-negative bacteria and induces various immune responses to infection. Recent studies have reported that LPS induces cellular stress in various cells including oocytes and embryos. Melatonin (N-acetyl-5-methoxytryptamine) is a regulatory hormone of circadian rhythm and a powerful antioxidant. It has been known that melatonin has an effective function in scavenging oxygen free radicals and has been used as an antioxidant to reduce the cytotoxic effects induced by LPS. However, the effect of melatonin on LPS treated early embryonic development has not yet been confirmed. In this study, we cultured mouse embryos in medium supplemented with LPS or/and melatonin up to the blastocyst stage in vitro and then evaluated the developmental rate. As a result of the LPS-treatment, the rate of blastocyst development was significantly reduced compared to the control group in all the LPS groups. Next, in the melatonin only treated group, there was no statistical difference in embryonic development and no toxic effects were observed. And then we found that the treatment of melatonin improved the rates of compaction and blastocyst development of LPS-treated embryos. In addition, we showed that melatonin treatment decreased ROS levels compared to the LPS only treated group. In conclusion, we demonstrated the protective effect of melatonin on the embryonic developmental rate reduced by LPS. These results suggest a direction to improve reproduction loss that may occur due to LPS exposure and bacterial infection through the using of melatonin during in vitro culture.

• The Korea Journal of Herbology
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• v.23 no.3
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• pp.103-112
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• 2008

Objectives : This research aimed to study the cytotoxicity and immuno modulating activity of fermented Artemisia argyi Lev. et Vant.(Compositae). Methods : Effect of fermented Artemisiae Argyi Folium extracts, which were fermented by Sacchromyces cerevisiae STV89(AFS), on cell viability, generation of ROS within cells, generation of NO and the level of cytokines($TNF-{\alpha}$ and IL-6) was measured using mouse macrophage RAW 264.7 cell. Results : 1. Result of MTT assay conducted to verify the cytotoxicity of fermented Artemisiae argyi folium extract illustrated that, when fermented Artemisiae argyi folium extract was processed for each concentration, there was no excessive induction of cytoxicity in the RAW 264.7 cell. 2. Fermented Artemisiae Argyi Folium extract increased the generation of H2O2 within RAW 264.7 cell as well as significantly increased inhibition of generation of H2O2 in macrophage induced by LPS. 3. Fermented Artemisiae Argyi Folium extract inhibited generation of NO in RAW 264.7 cell, and significantly inhibited increase in generation of NO of macrophage induced by LPS. 4. Fermented Artemisiae Argyi Folium extract, AFS has significantly reduced the increase in the generation of $TNF-{\alpha}$ above 10 ${\mu}g/mL$. 5. Fermented Artemisiae Argyi Folium extract, AFS has significantly reduced the increase in generation of IL-6 above 50 ${\mu}g/mL$. Conclusions : AFS fermented extract produced from Artemisiae Argyi Flium, have increased generation of ROS and reduced generation of NO in RAW 264.7 cell without excessively inducing cytotoxicity of RAW 264.7 cell. In addition, they displayed significant immuno modulating activities including inhibition of generation of $TNF-{\alpha}$ and IL-6 in macrophage, induced by LPS.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.2
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• pp.153-160
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• 2013

The peel from seven types of citrus was extracted with 80% methanol, and their phenolic compound content, oxygen radical absorbance capacity (ORAC), inhibitory activities of nitric oxide (NO), and reactive oxygen species (ROS) production induced by LPS and t-BHP in LPS-activated RAW 264.7 cells were measured. Total phenolic content was high in Yungkyool, Cheonhyehyang, and Jinkyool (30.6, 30.2, and 28.2 mg GAE/g, respectively), while total flavonoid content was high in Yungkyool and Jinkyool (30.3 and 25.5 mg RE/g, respectively). ORAC was the highest at 1,076 mM TE/g in Yungkyool, followed by Cheonhyehyang (1,012), Jinkyool (984), and Hallabong (914). High inhibitory activity against NO production was shown in Cheonhyehyang, Yungkyool, and Jinkyool with $IC_{50}$ values of 215.3, 259.2, and 328.9 ${\mu}g/mL$, respectively. LPS-induced ROS production was inhibited by 16.4% and 12.8% in Hallabong and Jinkyool, while t-BHP-induced ROS production was inhibited by 28.7%, 26.1%, and 26.6% in Jinkyool, Hallabong, and Cheonhyehyang, respectively. Correlation coefficients between total phenolic, total flavonoid content, and ORAC were 0.884 and 0.855. Inhibitory activity against NO production showed higher correlation with total phenolic content than total flavonoid content. It was concluded that citrus peels had potent antioxidant activities and could be used as natural antioxidants in the food and pharmaceutical industries.

• The Korea Journal of Herbology
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• v.25 no.1
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• pp.83-91
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• 2010

Objectives : In this study, we compared the anti-oxidant and anti-inflammatory activities of Curcumae longae Radix (CLRa) and Curcumae longae Rhizoma (CLRh). Methods : We performed 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and 2,2-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) cation scavenging assays, and determined total polyphenolic content to examine the anti-oxidant effects of CLRa and CLRh. We also evaluated the anti-oxidant effects of CLRa and CLRh against hydrogen peroxide ($H_2O_2$)-induced toxicity in PC12 cells using thiazolyl blue tetrazolium bromide (MTT) and reactive oxygen species (ROS) assays. Next, to compare the anti-inflammatory effects of CLRa and CLRh against lipopolysaccharide (LPS)-induced inflammation in microglia BV2 cells, we measured nitric oxide (NO) assay and inducible nitrite synthase (iNOS) using Western blotting analysis. Results : CLRa showed higher activity in DPPH and ABTS assays and lower total polyphenolic contents compared with CLRh. In PC12 cells, CLRa and CLRh showed no difference in H2O2-induced cell toxicity and ROS overproduction. In BV2 cells, CLRa showed higher effect than CLRh in NO and iNOS production induced by LPS. Conclusions : These results demonstrate that CLRa has higher radical scavenging activities and anti-inflammatory effect in BV2 cells comparing CLRh. However, CLRa and CLRh have no effect and no difference in $H_2O_2$-induced toxicity.

• Microbiology and Biotechnology Letters
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• v.50 no.1
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• pp.22-30
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• 2022

Haliotis discus hannai called abalone, is the valuable marine mollusks and the by-products of abalone processing are viscera. Brownish abalone male viscera (AMV), which have not been reported as having anti-inflammatory effects, was extracted with acetone and fractionated by different six acetone/hexane ratios (0, 10, 20, 30, 40, and 100%) using a silica column via in vitro ABTS and DPPH radical and nitric oxide (NO) production assay-guided fractionation. Among the fractions, the acetone/hexane ratio 40%, A40 exhibited the most potent radical scavenging activities and inhibition of lipopolysaccharide (LPS)-induced NO production without cytotoxicity. A40 inhibited LPS-induced intracellular reactive oxygen species (ROS) production in a dose-dependent manner. Western blot analysis revealed that A40 down-regulated the activation of NF-κB, MAPK (ERK 1/2, p-38, and JNK), and inflammatory enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2. Moreover, this fraction inhibited the generation of pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. These results suggested that AMV containing A40 with anti-inflammatory and anti-oxidantive effects, is the effective therapeutic and functional material for treating inflammatory disorders.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.6
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• pp.1320-1331
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• 2009

Oxidative stress and inflammation are important events in the development of chronic inflammatory diseases including arthritis, atherosclerosis, diabetes, hypertension. Cynomorium songaricum (CS) has been used as a traditional Korean herbal medicine, and it is currently used in traditional clinics to treat frequent urination, spermatorrhea, weakness of the sinews and constipation in the folk medicine. The aim of this study was to determine whether fractionated extracts of CS inhibit free radical generation such as DPPH radical, superoxide radical, nitric oxide and peroxynitrite, production of nitrite an index of NO, $PGE_2$, iNOS, COX-2 and pro-inflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Our results indicated that the most superior extract which scavenged DPPH radical, reactive oxygen species (ROS) and RNS was CS ethyl acetate extract (CSEA). Moreover, CSEA significantly inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of TNF-$\alpha$, IL-$1{\beta}$ and IL-6 formation in macrophages. Furthermore, CSEA treatment also blocked LPS-induced intracellular ROS production and the activation of NF-${\kappa}B$. These findings indicate that CSEA inhibits the production of pro-inflammatory mediators and cytokines via the suppression of ROS production and NF-${\kappa}B$ activation. Take together, these results indicate that CSEA has the potential for use as an natural anti-oxidant and an agent of anti-chronic inflammatory diseases.

• The Korea Journal of Herbology
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• v.30 no.4
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• pp.81-88
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• 2015

Objectives : The aim of present study was to evaluate the beneficial effect of Scutellariae Radix (SR) and Scutellariae Radix EtOH-heated at 200℃ (SR200) using lipopolysaccharide (LPS) treated intestine of mice.Methods : Extract of SR and SR200 were orally administrated. Their effects were compared with vehicletreated LPS and normal groups. Subsequently, we measured reactive oxygen species (ROS) and nitric oxide in the serum and western blotting in the intestine.Results : The average weight in LPS treated (Vehicle) group was lowered significantly compare to that in non-treated normal group and this weight loss in the vehicle group was effectively prevented by the administration of SR and SR200 respectively. The increased oxidative stress biomarker levels such as reactive oxygen species (ROS) and nitric oxide (NO) in the serum was markedly decreased by treated with SR200. The decreased levels of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) induced by LPS injection were significantly restored by both SR and SR200 treatment. Moreover, increased inflammatory mediators and cytokines such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) in the LPS treated vehicle mice were significantly decreased through down-regulation c-JUN through reduction of oxidative stress.Conclusions : SR and SR200 could have benefit effect through down-regulation of abnormal oxidative stress in LPS induced intestine injury mice. Moreover, The anti-inflammatory activity of SR200 extract was better than SR extract in the LPS induced intestine injury mice.

• Korean Journal of Medicinal Crop Science
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• v.17 no.1
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• pp.8-14
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• 2009

Pueraria lobata (Willd.) Ohwi was investigated to check its modulatory effects on the activation of macrophages upon inflammatory conditions treatment. For this purpose, we examined several inflammatory responses such as nitric oxide (NO) production, reactive oxygen species (ROS) generation, cytoprotection and phagocytosis under the treatment of methanol extract from P. lobata (Pl-ME). Pl-ME dose-dependently blocked NO production in lipopolysaccharide (LPS)- stimulated RAW264.7 cells but not sodium prusside (SNP)-generated NO release. The NO inhibition seemed to be due to blocking inducible NO synthase (iNOS), since Pl-ME suppressed its expression in a NF-${\kappa}B$-independent manner. Similarly, this extract also effectively protected RAW264.7 cells from LPS-induced cytotoxicity. However, Pl-ME did not block ROS generation and rather it enhanced. Finally, this extract negatively modulated FITC-dextran uptake. Therefore, our data suggested that Pl-ME may be involved in negatively regulating some macrophage-mediated inflammatory responses such as NO production and phagocytic uptake.

• The Korea Journal of Herbology
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• v.26 no.2
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• pp.17-23
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• 2011

Objectives : This study aims at examining the effect of the fermentative extract of root of Sophorae Radix on the immuno-modulating activity. Methods : Cell viabilities were measured by MTT assay. Effect of SFS on nitric oxide(NO), hydrogen peroxide production from RAW 264.7 cells was accessed by Griess reagent assay. Effect of SFS on productions of inflammatory cytokines such as TNF-${\alpha}$, IL-6 in LPS-induced RAW 264.7 cells was accessed by a multiplex bead array assay based on xMAP technology. Results : The results of the experiment are as follows. 1. As a result of carrying out MTT assay to check the cellular toxicity of the fermentative extract of Sophorae Radix. There was not any excessive toxicity to the macrophage when the fermentative extract of root of Sophorae Radix was treated in different concentrations. 2. The fermentative extract of Sophorae Radix increased the generation of hydrogen peroxide in the macrophage and significantly restored the suppression of the generation of the hydrogen peroxide in the macrophage induced by LPS. 3. The fermentative extract of Sophorae Radix reduced the generation of NO in the macrophage and significantly suppressed the increase of the generation of NO in the macrophage induced by LPS. 4. The fermentative extract of Sophorae Radix significantly decreased the amount of TNF-${\alpha}$ generated in the macrophage induced by LPS when it was $25{\mu}g/mL$ or higher. Conclusion : These results suggest that SFS has anti-inflammatory moiety related with its inhibition of NO, hydrogen peroxide, TNF-${\alpha}$, IL-6, in macrophage led by LPS.

• Journal of Life Science
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• v.21 no.10
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• pp.1377-1384
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• 2011

Astaxanthin (ATX) is a red-orange carotenoid pigment that occurs naturally in a wide variety of living organisms. In this study we investigated the inhibitory effects of ATX on the induction of inducible nitric oxide synthase (iNOS), nitric oxide (NO), proinflammatory cytokines, nuclear factor-kappa B(NF-${\kappa}B$) and reactive oxygen species (ROS) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. In addition, we tested the superoxide radical scavenging activity of ATX by scavenging assay. iNOS and NF-${\kappa}B$ expressions were determined by immunoblot analysis. Interleukin (IL)-6 and tumour necrosis factor-${\alpha}$ (TNF-${\alpha}$) were assayed by ELISA. NO production was monitored by measuring the amount of nitrite. ROS was examined by using the 2', 7'-Dichlorodihydrofluorescin diacetate (DCFH-DA) method. At a concentration of 100 ${\mu}M$, ATX inhibited the expression level of LPS-induced NF-${\kappa}B$, as well as the production of LPS-induced NO and proinflammatory cytokines (IL-6 and TNF-${\alpha}$), by suppressing iNOS expression. In particular, the maximal inhibition rate of IL-6 and TNF-${\alpha}$ production by ATX (100 ${\mu}M$) was 65.2----- and 21.2-----, respectively. In addition, ATX inhibited the LPS-induced transcriptional activity of NF-${\kappa}B$, and this was associated with suppressing the translocations of NF-${\kappa}B$ from the cytosol to the nucleus. Moreover, at various concentrations (25-100 ${\mu}M$), ATX inhibited the intracellular level of ROS. At a concentration of 5 mg/ml, the superoxide radical scavenging activity of ATX was 1.33 times higher than ${\alpha}$-tocopherol of the same concentration. These results showed that ATX inhibited the expression of iNOS and the production of NO and proinflammatory cytokines resulting from ROS production and NF-${\kappa}B$ activation in macrophages. Furthermore, ATX was found to be more effective in superoxide radical scavenging activities compared to ${\alpha}$-tocopherol. These findings are expected to strengthen the position of ATX as anti-inflammatory medicine and antioxidant.