• Title/Summary/Keyword: LPS-induced ROS and NO

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Eryngium foetidum Suppresses Inflammatory Mediators Produced by Macrophages

  • Mekhora, Chusana;Muangnoi, Channarong;Chingsuwanrote, Pimjai;Dawilai, Suwitcha;Svasti, Saovaros;Chasri, Kaimuk;Tuntipopipat, Siriporn
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.2
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    • pp.653-664
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    • 2012
  • Objective: This study assessed anti-inflammatory and antioxidant activities of $E.$ $foetidum$ leaf extract on LPS-activated murine macrophages. Methods: RAW264.7 cells were pretreated with or without $E.$ $foetidum$ extract for 1 h prior to incubation with LPS for 24 h. Anti-inflammatory activity was evaluated with reference to iNOS, COX-2, TNF-${\alpha}$ and IL-6 gene expression. In addition, NO and intracellular ROS generation were determined by Griess method and fluorescence intensity and activation of MAPKs and $I{\kappa}B$ by Western blotting. Results: Prior treatment with $E.$ $foetidum$ leaf extract inhibited elevation of IL-6, TNF-${\alpha}$, iNOS and COX-2, together with their cognate mRNAs in a dose-dependent manner. NO and intracellular ROS contents were similarly reduced. These effects were due to inhibition of LPS-induced phosphorylation of JNK and p38 as well as $I{\kappa}B$. $E.$ $foetidum$ ethanol extract were shown to contain lutein, ${\beta}$-carotene, chlorogenic acid, kaempferol and caffeic acid, compounds known to exert these bioactive properties. Conclusions: $E.$ $foetidum$ leaf extract possesses suppressive effects against pro-inflammatory mediators. Thus, $E.$ $foetidum$ has a high potential to be used as a food supplement to reduce risk of cancer associated with inflammation.

A Screen for Dual-protection Molecules from a Natural Product Library against Neuronal Cell Death and Microglial Cell Activation (신경세포 사멸과 미세아교세포활성화 억제 동시 가능 천연물질 탐색 연구)

  • Min, Ju-Sik;Lee, Dong-Seok
    • Journal of Life Science
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    • v.25 no.6
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    • pp.656-662
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    • 2015
  • Natural products and natural product structures play a general and highly significant role in drug discovery and development process because it has various merits and potentials for new drug source that have extensive clinical experience, development time contraction, excellent stability and safety. In several neurological disorders, neuronal death and excessive activation of microglia (neuro-inflammation) are observed. A number of drug discovery-related neuronal cell death and neuro-inflammation was studied from natural products, respectively. However, until now, it has not been possible to study dual-protection molecules recorded in the Natural Product library. In the present study, using the natural product-derived library of the Institute for Korea Traditional Medical Industry, we investigated dual-protective molecules against glutamate (a classical excitatory neurotransmitter)-induced oxidative stress mediated neuronal cell death and LPS-induced excessive activated microglial cells (immune cells of the brain). Chrysophanol, extracted from Rheum palmatum, had dual-protective effects against both glutamate-induced neuronal cell death and LPS-induced NO production, triggering proinflammatory cytokines and microglia activation and resulting in neuroinflammation. Flow-cytometry analysis revealed that chrysophanol had a scavenger effect, scavenging glutamate- and LPS-induced reactive oxygen species (ROS) produced by neuronal and microglial cells, respectively. Based on the present study, chrysophanol may have an important protective role against neuronal cell death and neuroinflammation in the brain. The results may be helpful for studying drug development candidates for treating central nervous system disorders.

Orostachys japonicus Hexane Fraction Attenuates Pro-inflammatory Cytokines in LPS-activated Macrophage Cells by Suppression of AP-1 and IRF3 Transcription Factors (LPS로 유도된 대식세포에 대한 와송 핵산추출물의 AP-1과 IRF3 전사인자의 억제에 의한 전염증성 사이토카인의 감소 효과)

  • Lee, Hyeong-Seon
    • Microbiology and Biotechnology Letters
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    • v.48 no.3
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    • pp.310-315
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    • 2020
  • Orostachys japonicus (O. japonicus) is known as a medicinal plant for the treatment of various symptoms. This study investigated the anti-inflammatory effect of the hexane fraction from O. japonicus (OJH) on the LPS-stimulated response in RAW 264.7 macrophage cells. This study was conducted to confirm the effect of cell cytotoxicity and production of reactive oxygen species (ROS) in OJH-treated macrophage cells. Additionally, pro-inflammatory cytokines and transcription factors were determined using RT-PCR and western blotting assay. OJH showed no change in lactate dehydrogenase (LDH) levels and exhibited reduced ROS levels in LPS-induced inflammatory cells. Moreover, OJH significantly suppressed the mRNA levels of proinflammatory cytokines, including IL-1β, IL-2, IL-6, TNF-α, and IP-10. Furthermore, OJH effectively inhibited the protein levels of AP-1 (p-c-Jun and p-c-Fos) and p-IRF3 in a dose-dependent manner. In conclusion, our results demonstrate that OJH exhibits strong anti-inflammatory activities via regulation of inflammatory factors.

Inhibitory Effect of Ginsenoside-Rp1, a Novel Ginsenoside Derivative, on the Functional Activation of Macrophage-like Cells

  • Park, Tae-Yoon;Cho, Jae-Youl
    • Biomolecules & Therapeutics
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    • v.16 no.4
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    • pp.370-376
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    • 2008
  • Ginsenoside Rp1 (G-Rp1) is a ginseng saponin derivative with chemopreventive and anti-cancer activities. In this study, we examined the regulatory activity of G-Rp1 on the functional activation of macrophages. G-Rp1 remarkably inhibited TNF-$\alpha$ production, LPS-induced cell cytotoxicity, NO production, ROS generation, and phagocytic uptake from lipopolysacchride (LPS)-activated RAW264.7 cells. According to structural feature study using several G-Rp1 analogs, two carbohydrates (glucose-glucose) at R1 position were observedto be highly effective, compared to other structural derivatives. Although the inhibitory activities of G-Rp1 on macrophage functions were not remarkable, several points that G-Rp1 was known to be safe, and that this compound was orally effective, suggest that G-Rp1 may be beneficial in treating macrophage-mediated immunological diseases.

Luteolin 5-O-glucoside from Korean Milk Thistle, Cirsium maackii, Exhibits Anti-Inflammatory Activity via Activation of the Nrf2/HO-1 Pathway

  • Jung, Hyun Ah;Roy, Anupom;Abdul, Qudeer Ahmed;Kim, Hyeung Rak;Park, Hee Juhn;Choi, Jae Sue
    • Natural Product Sciences
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    • v.23 no.3
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    • pp.183-191
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    • 2017
  • Luteolin 5-O-glucoside is the major flavonoid from Korean thistle, Cirsium maackii. We previously reported the anti-inflammatory activities of luteolin 5-O-glucoside in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. In this study, we determined the anti-inflammatory mechanisms of luteolin 5-O-glucoside through the inhibition of nitric oxide (NO) production in vitro and in vivo. Results revealed that luteolin 5-O-glucoside dose-dependently inhibited NO production and expression of iNOS and COX-2 in LPS-induced RAW 264.7 cells. Luteolin 5-O-glucoside also significantly inhibited the translocation of $NF-{\kappa}B$, the activation of MAPKs, and ROS generation in LPS-induced RAW 264.7 cells. In addition, protein expressions of Nrf-2 and HO-1 were also upregulated by luteolin 5-O-glucoside treatment. Moreover, luteolin 5-O-glucoside inhibited ${\lambda}-carrageenan-induced$ mouse paw edema by 65.34% and 48.31% at doses of 50 and 100 mg/kg body weight, respectively. These findings indicate potential anti-inflammatory effect of luteolin 5-O-glucoside particularly by downregulating $NF-{\kappa}B$ and upregulating HO-1/Nrf-2 pathway.

Anti-oxidative and Anti-inflammatory Activities of Desmodium heterocarpon Extract in RAW 264.7 Cells (RAW 264.7 세포에서 Desmodium heterocarpon 추출물의 항산화 및 항염증 활성)

  • Lee, Su Hyeon;Jin, Kyong-Suk;Son, Yu Ri;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.28 no.2
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    • pp.216-222
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    • 2018
  • Desmodium heterocarpon is one of vines belongs to Fabaceae family, mainly distributed in Asian countries such as Korea and Japan. This study was conducted to explore new nutraceutical resources from the plant kingdom possessing biological activities. To fulfill this purpose, the anti-oxidative and anti-inflammatory activities of D. heterocarpon ethanol extract (DHEE) were evaluated by 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity assay, reactive oxygen species (ROS) scavenging activity assay, nitric oxide (NO) inhibitory activity assay, and the analysis of related protein expressions by Western blot hybridization. DHEE exhibited potent anti-oxidative activity as confirmed by DPPH radical scavenging capacity against DPPH similar with ascorbic acid, a well-known anti-oxidative agent, used as a positive control. DHEE also effectively suppressed hydrogen peroxide ($H_2O_2$)-induced ROS on RAW 264.7 murine macrophage cells. Furthermore, DHEE induced the expression of the anti-oxidative enzyme heme oxygenase 1 (HO-1), and its upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2) as a dose dependent manner. DHEE inhibited lipopolysaccharide (LPS) induced nitric oxide (NO) formation as a consequence of inducible NO synthase (iNOS) down regulation. Taken together, these results suggest that DHEE has anti-oxidative and anti-inflammatory activities and thus appears to be useful sources as potential anti-oxidant and anti-inflammatory agents. The identification of active compounds that confer biological activities of DHEE might be needed.

Comparative study of the biological activities effect of Mentha arvensis L. extracts from water and 80% ethanol (열수 및 80% 에탄올로 추출한 박하의 생리활성 비교연구)

  • Choi, Dong-Hun;Seung, O-Tak;Lim, Mi-Hye
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.1
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    • pp.208-216
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    • 2019
  • The purpose of this study was to investigated the biological activities such as cytotoxic, anti-oxidant and anti-inflammatory using Mentha arvensis L. extracts. Mentha arvensis L. was prepared by extracting with DW and 80% ethanol, after cell viability was assessed by MTT assay using RAW 264.7 cells. Antioxidant activities, and Anti-inflammatory activities measured through changes in the levels of reactive oxygen species (ROS), nitric oxide (NO), leukotrien B4 (LTB4), and anti- or in-flammatory cytokines($IL-1{\beta}$, IL-6, tumor necrosis factor $(TNF)-{\alpha}$, and IL-10) on LPS-induced RAW 264.7 cells. All test measured by ELISA reader and Luminex. Mentha arvensis L. was no cytotoxic in water and 80% ethanol extracts, and Concentration of 100 ug/ml of 80% ethanol extract was suppressed the productions of ROS in LPS-induced RAW 264.7 cells. Also, Productions of NO, LTB4, inflammatory or anti-inflammatory cytokines showed efficacy change that dose-independent of all extracts. In other words, Mentha arvensis L. showed significant biological activities showing anti-inflammatory, and anti-oxidant. These results may be developed as a raw material for new health food and therapeutics to ease the symptoms mentioned above.

Anti-oxidative and Anti-inflammatory Activities of Decaisnea insignis Ethanol Extract (Decaisnea insignis 에탄올 추출물의 항산화 및 항염증 활성)

  • Jin, Kyong-Suk;Lee, Ji Young;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.24 no.9
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    • pp.973-980
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    • 2014
  • This study was conducted to explore new nutraceutical resources from the plant kingdom possessing biological activities. To fulfill this purpose, the anti-oxidative and anti-inflammatory activities of Decaisnea insignis ethanol extract (DIEE) were evaluated. First, DIEE possessed potent scavenging activity against 1,1-diphenyl-2-picryl hydrazyl (DPPH), similar to ascorbic acid used as a positive control. Moreover, DIEE inhibited lipopolysaccharide (LPS)- and hydrogen peroxide ($H_2O_2$)-induced reactive oxygen species (ROS) in RAW 264.7 cells. Furthermore, DIEE induced the expression of an anti-oxidative enzyme, heme oxygenase 1 (HO-1), and its upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2), in a dose-dependent manner. The modulation of the HO-1 and Nrf2 expressions might be regulated by mitogen-activated protein kinases (MAPKs) and their upstream signaling pathways. On the other hand, DIEE suppressed LPS-induced nitric oxide (NO) formation without cytotoxicity. The inhibition of the NO formation was the result of the downregulation of inducible NO synthase (iNOS) by DIEE. The suppression of NO and iNOS by DIEE might be modulated by their upstream transcription factors, nuclear factor ${\kappa}B$ ($NF-{\kappa}B$), and activator protein 1 (AP-1) pathways. Taken together, these results provide important new insights that D. insignis possesses anti-oxidative and anti-inflammatory activities. Therefore, it might be utilized as a promising material in the field of nutraceuticals.

Establishment of inflammatory model induced by Pseudorabies virus infection in mice

  • Ren, Chun-Zhi;Hu, Wen-Yue;Zhang, Jin-Wu;Wei, Ying-Yi;Yu, Mei-Ling;Hu, Ting-Jun
    • Journal of Veterinary Science
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    • v.22 no.2
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    • pp.20.1-20.13
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    • 2021
  • Background: Pseudorabies virus (PRV) infection leads to high mortality in swine. Despite extensive efforts, effective treatments against PRV infection are limited. Furthermore, the inflammatory response induced by PRV strain GXLB-2013 is unclear. Objectives: Our study aimed to investigate the inflammatory response induced by PRV strain GXLB-2013, establish an inflammation model to elucidate the pathogenesis of PRV infection further, and develop effective drugs against PRV infection. Methods: Kunming mice were infected intramuscularly with medium, LPS, and different doses of PRV-GXLB-2013. Viral spread and histopathological damage to brain, spleen, and lung were determined at 7 days post-infection (dpi). Immune organ indices, levels of reactive oxygen species (ROS), nitric oxide (NO), and inflammatory cytokines, as well as levels of activity of COX-2 and iNOS were determined at 4, 7, and 14 dpi. Results: At 105-106 TCID50 PRV produced obviously neurological symptoms and 100% mortality in mice. Viral antigens were detectable in kidney, heart, lung, liver, spleen, and brain. In addition, inflammatory injuries were apparent in brain, spleen, and lung of PRV-infected mice. Moreover, PRV induced increases in immune organ indices, ROS and NO levels, activity of COX-2 and iNOS, and the content of key pro-inflammatory cytokines, including interleukin (IL)-1β, IL-6, tumor necrosis factor-α, interferon-γ and MCP-1. Among the tested doses, 102 TCID50 of PRV produced a significant inflammatory mediator increase. Conclusions: An inflammatory model induced by PRV infection was established in mice, and 102 TCID50 PRV was considered as the best concentration for the establishment of the model.

Pinus Densiflora Gnarl Extract for Pharmacopuncture Inhibits Inflammatory Responses through Heme Oxygenase-1 Induction in Lipopolysaccharide-Stimulated RAW264.7 Macrophages (RAW 264.7 세포에서 Heme Oxygenase-1 발현 유도를 통한 송절(松節) 약침액의 항염증 효능)

  • Lee, Kang-Pa;Moon, Jin-Young
    • Korean Journal of Acupuncture
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    • v.29 no.1
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    • pp.37-46
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    • 2012
  • Objectives : The gnarl of Pinus densiflora, called Songjeol in Korea, has been used as a medicinal herb for the treatment of inflammatory-related diseases such as arthralgia, myalgia and bruise. However, the molecular actions and mechanisms have not been clearly investigated. The aim of this study was to clarify the anti-inflammatory activity of Pinus densiflora gnarl pharmacopuncture (PDGP) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Methods : Cytotoxicity was assessed by XTT assay. The amount of nitric oxide (NO) production was determined by nitrite assay. The mRNA expressions of interleukin-$1{\beta}$ (IL-$1{\beta}$), interleukin-6 (IL-6), cyclooxygenase-2 (COX-2) and heme oxygenase-1 (HO-1) were analyzed by RT-PCR. Reactive oxidative species (ROS) generation was measured using the fluorescence microscopy. In addition, inducible nitric oxide synthase (iNOS) and redox factor-1 (Ref-1) protein expressions were detected by Western blotting. Results : PDGP inhibited NO production and ROS generation in LPS-stimulated RAW264.7 cells. At the mRNA level, PDGP suppressed IL-$1{\beta}$, IL-6 and COX-2 expression. On the other hand, PDGP induced HO-1 mRNA expression. Furthermore, PDGP suppressed iNOS and Ref-1 protein expression. Conclusions : This result suggests that PDGP can act as a suppressor agent on NO and iNOS through induction of HO-1, and play an useful role in blocking inflammatory responses.