• Title/Summary/Keyword: raw 264.7 macrophages

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Calcium-doped zinc oxide nanocrystals as an innovative intracanal medicament: a pilot study

  • Gabriela Leite de Souza;Thamara Eduarda Alves Magalhaes;Gabrielle Alves Nunes Freitas;Nelly Xiomara Alvarado Lemus;Gabriella Lopes de Rezende Barbosa;Anielle Christine Almeida Silva;Camilla Christian Gomes Moura
    • Restorative Dentistry and Endodontics
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    • v.47 no.4
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    • pp.38.1-38.15
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    • 2022
  • Objectives: This study investigated the cytotoxicity, radiopacity, pH, and dentinal tubule penetration of a paste of 1.0% calcium-doped zinc oxide nanocrystals (ZnO:1.0Ca) combined with propylene glycol (PRG) or polyethylene glycol and propylene glycol (PEG-PRG). Materials and Methods: The pastes were prepared by mixing calcium hydroxide [Ca(OH)2] or ZnO:1.0Ca with PRG or a PEG-PRG mixture. The pH was evaluated after 24 and 96 hours of storage in deionized water. Digital radiographs were acquired for radiopacity analysis and bubble counting of each material. The materials were labeled with 0.1% fluorescein and applied to root canals, and images of their dentinal tubule penetration were obtained using confocal laser scanning microscopy. RAW264.7 macrophages were placed in different dilutions of culture media previously exposed to the materials for 24 and 96 hours and tested for cell viability using the MTT assay. Analysis of variance and the Tukey test (α = 0.05) were performed. Results: ZnO:1.0Ca materials showed lower viability at 1:1 and 1:2 dilutions than Ca(OH)2 materials (p < 0.0001). Ca(OH)2 had higher pH values than ZnO:1.0Ca at 24 and 96 hours, regardless of the vehicle (p < 0.05). ZnO:1.0Ca pastes showed higher radiopacity than Ca(OH)2 pastes (p < 0.01). No between-material differences were found in bubble counting (p = 0.0902). The ZnO:1.0Ca pastes had a greater penetration depth than Ca(OH)2 in the apical third (p < 0.0001). Conclusions: ZnO:1.0Ca medicaments presented higher penetrability, cell viability, and radiopacity than Ca(OH)2. Higher values of cell viability and pH were present in Ca(OH)2 than in ZnO:1.0Ca.

Anti-inflammation and Anti-cancer Activity of Methanol Extract of Antarctic Lichen, Usnea Aurantiaco-atra (남극 지의류 Usnea Aurantiaco-atra의 메탄올 추출물의 항염증 및 항암 활성)

  • Sung-Suk Suh
    • Journal of Life Science
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    • v.33 no.12
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    • pp.978-986
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    • 2023
  • Inflammation by the innate immune system is a protective mechanism of the organism against infection-mediated environmental factors. It is also responsible for the pathogenesis of various human diseases, including the progression of cancer. Lichens are receiving increasing attention as a source of bioactive molecules with therapeutic potential for a variety of diseases. Additionally, the antioxidant, anti-inflammatory, and anticancer potential of lichen and its secondary metabolites have been widely reported. However, the underlying mechanism is still unknown. In the present study, to investigate molecular mechanisms of anti-inflammation and anti-cancer activity in the Antarctic lichen, Usnea aurantiaco-atra, methanol extract of Usnea aurantiaco-atra (MEUS) was used in vitro assays in RAW 264.7 macrophages cell and HCT116 colon cancer cells. Based on our data, MEUS had the anti-inflammatory activity through the modulation of main inflammatory indicators such as COX-2, IL-6, iNOS, TNF-α and NO production in a concentration-dependent manner. In addition, we observed that MEUS had cytotoxic activity against HCT116 colon cancer cells in a concentration-dependent manner, leading to a significantly reduced proliferation of the cancer cells through apoptotic induction by activating caspase-3. Taken together, this work firstly reported the anti-inflammatory and anti-cancer activities of an Antarctic lichen, Usnea aurantiaco-atra, and MEUS may provide a new insight into the molecular mechanisms underlying a link between inflammation and cancer.

Amelioration of colitis progression by ginseng-derived exosome-like nanoparticles through suppression of inflammatory cytokines

  • Jisu Kim;Shuya Zhang ;Ying Zhu;Ruirui Wang;Jianxin Wang
    • Journal of Ginseng Research
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    • v.47 no.5
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    • pp.627-637
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    • 2023
  • Background: Damage to the healthy intestinal epithelial layer and regulation of the intestinal immune system, closely interrelated, are considered pivotal parts of the curative treatment for inflammatory bowel disease (IBD). Plant-based diets and phytochemicals can support the immune microenvironment in the intestinal epithelial barrier for a balanced immune system by improving the intestinal microecological balance and may have therapeutic potential in colitis. However, there have been only a few reports on the therapeutic potential of plant-derived exosome-like nanoparticles (PENs) and the underlying mechanism in colitis. This study aimed to assess the therapeutic effect of PENs from Panax ginseng, ginseng-derived exosome-like nanoparticles (GENs), in a mouse model of IBD, with a focus on the intestinal immune microenvironment. Method: To evaluate the anti-inflammatory effect of GENs on acute colitis, we treated GENs in Caco2 and lipopolysaccharide (LPS) -induced RAW 264.7 macrophages and analyzed the gene expression of proinflammatory cytokines and anti-inflammatory cytokines such as TNF-α, IL-6, and IL-10 by real-time PCR (RT-PCR). Furthermore, we further examined bacterial DNA from feces and determined the alteration of gut microbiota composition in DSS-induced colitis mice after administration of GENs through 16S rRNA gene sequencing analysis. Result: GENs with low toxicity showed a long-lasting intestinal retention effect for 48 h, which could lead to effective suppression of pro-inflammatory cytokines such as TNF-α and IL-6 production through inhibition of NF-κB in DSS-induced colitis. As a result, it showed longer colon length and suppressed thickening of the colon wall in the mice treated with GENs. Due to the amelioration of the progression of DSS-induced colitis with GENs treatment, the prolonged survival rate was observed for 17 days compared to 9 days in the PBS-treated group. In the gut microbiota analysis, the ratio of Firmicutes/Bacteroidota was decreased, which means GENs have therapeutic effectiveness against IBD. Ingesting GENs would be expected to slow colitis progression, strengthen the gut microbiota, and maintain gut homeostasis by preventing bacterial dysbiosis. Conclusion: GENs have a therapeutic effect on colitis through modulation of the intestinal microbiota and immune microenvironment. GENs not only ameliorate the inflammation in the damaged intestine by downregulating pro-inflammatory cytokines but also help balance the microbiota on the intestinal barrier and thereby improve the digestive system.

Antioxidant and Immunological Activities of Polysaccharide Extracted from Cultured Mycelia of Schizophyllum commune (치마버섯 균사체 배양물로부터 분리한 다당류의 항산화 및 면역 활성)

  • Lee, June-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.9
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    • pp.1334-1341
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    • 2014
  • To examine the biological activity of polysaccharide extracted from cultured mycelia of Schizophyllum commune, we determined anti-complementary activity and nitric oxide production as a measure of immunological activity, anti-lipidperoxidation and hydroxy radical scavenging activity as a measure of antioxidative activity, tyrosinase inhibitory activity, anti-microbial activity, and transdermal flux of polysaccharide extracted from cultured mycelia of S. commune. Polysaccharide extracted from S. commune activated the complementary system and produced nitric oxide in RAW 264.7 macrophages. Antioxidant activities as malondialdehyde values were $49.5{\pm}0.7$, $39.7{\pm}1.7$, $39.2{\pm}1.2$, and $2.6{\pm}0.5nM/mL$ for control, extracellular polysaccharide extracted from S. commune (SC-EP), ultrafiltrated polysaccharide extracted from S. commune (SC-UP), and butylated hydroxytoluene, respectively. Hydroxy radical scavenging activity ($IC_{50}$) of SC-UP and mannitol were 3.32 and 1.66 mg/mL, respectively. Tyrosinase inhibitory activities of SC-UP, arbutin, and kojic acid were 19.9%, 31.8%, and 99.0%, respectively. Anti-microbial activities of SC-UP appeared to be low, and transdermal fluxes of SC-UP were 0.47%, 0.73%, and 1.20% after 3, 6, and 9 hr, respectively. These findings suggest that polysaccharide extracted from S. commune has potential immunological and antioxidant activities.

Oxidative Inactivation of Peroxiredoxin Isoforms by H2O2 in Pulmonary Epithelial, Macrophage, and other Cell Lines with their Subsequent Regeneration (폐포상피세포, 대식세포를 비롯한 각종 세포주에서 H2O2에 의한 Peroxiredoxin 동위효소들의 산화에 따른 불활성화와 재생)

  • Oh, Yoon Jung;Kim, Young Sun;Choi, Young In;Shin, Seung Soo;Park, Joo Hun;Choi, Young Hwa;Park, Kwang Joo;Park, Rae Woong;Hwang, Sung Chul
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.1
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    • pp.31-42
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    • 2005
  • Background : Peroxiredoxins (Prxs) are a relatively newly recognized, novel family of peroxidases that reduce $H_2O_2$ and alkylhydroperoxide into water and alcohol, respectively. There are 6 known isoforms of Prxs present in human cells. Normally, Prxs exist in a head-to-tail homodimeric state in a reduced form. However, in the presence of excess $H_2O_2$, it can be oxidized on its catalytically active cysteine site into inactive oxidized forms. This study surveyed the types of the Prx isoforms present in the pulmonary epithelial, macrophage, endothelial, and other cell lines and observed their response to oxidative stress. Methods : This study examined the effect of exogenous, excess $H_2O_2$ on the Prxs of established cell lines originating from the pulmonary epithelium, macrophages, and other cell lines, which are known to be exposed to high oxygen partial pressures or are believed to be subject to frequent oxidative stress, using non-reducing SDS polyacrylamide electrophoresis (PAGE) and 2 dimensional electrophoresis. Result : The addition of excess $H_2O_2$ to the culture media of the various cell-lines caused the immediate inactivation of Prxs, as evidenced by their inability to form dimers by a disulfide cross linkage. This was detected as a subsequent shift to its monomeric forms on the non-reducing SDS PAGE. These findings were further confirmed by 2 dimensional electrophoresis and immunoblot analysis by a shift toward a more acidic isoelectric point (pI). However, the subsequent reappearance of the dimeric Prxs with a comparable, corresponding decrease in the monomeric bands was noted on the non-reducing SDS PAGE as early as 30 minutes after the $H_2O_2$ treatment suggesting regeneration after oxidation. The regenerated dimers can again be converted to the inactivated form by a repeated $H_2O_2$ treatment, indicating that the protein is still catalytically active. The recovery of Prxs to the original dimeric state was not inhibited by a pre-treatment with cycloheximide, nor by a pretreatment with inhibitors of protein synthesis, which suggests that the reappearance of dimers occurs via a regeneration process rather than via the de novo synthesis of the active protein. Conclusion : The cells, in general, appeared to be equipped with an established system for regenerating inactivated Prxs, and this system may function as a molecular "on-off switch" in various oxidative signal transduction processes. The same mechanisms might applicable other proteins associated with signal transduction where the active catalytic site cysteines exist.

Nutritional Composition and Biological Activities of the Methanol Extracts of Sea Mustard (Undaria pinnatifida) in Market. (시판 미역의 영양성분 및 생리활성 분석)

  • Choi, Jae-Suk;Bae, Hee-Jung;Kim, Yang-Chun;Park, Nam-Hee;Kim, Tae-Bong;Choi, Young-Ju;Choi, Eun-Young;Park, Sun-Mee;Choi, In-Soon
    • Journal of Life Science
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    • v.18 no.3
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    • pp.387-394
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    • 2008
  • This research was performed to determine the proximate compositions, mineral contents, alginic acid, antioxidative activities and amino acids of sea mustards (Miyeok: Undaria pinnatifida) collected from Gijang and Wando area. Ash content was higher in Gijang samples, whereas carbohydrate and moisture were higher in Wando Sil Miyeok. General compositions of dried sea mustard showed different contents as manufacture's company and places. The major free amino acids were hydroxyproline, alanine, glutamic acid and asparagine in Gijang samples. Both Gijang and Wando Sil Miyeok showed lower contents comparing with Gijang Gadak Miyeok. Major mineral content was Na, K, Ca, Mg and P, and especially, Na and K were the most abundant in both Gijang and Wando sea mustards. Alginic acid content was almost similar in both sea mustards. Antioxidative activity of methanol extract of sea mustards was measured by using DPPH radical scavenging and SOD-like activity. DPPH radical scavenging activity was 45.5% $(40\;{\mu}g/ml)$ in Gijang Gadak Miyeok and 37.0% and 26.0% $(40\;{\mu}g/ml)$ in Gijang and Wando Sil Miyok, respectively. SOD-like activity of Gijang and Wando Sil Miyok was 63% and 71% $(10\;{\mu}g/ml)$, respectively. These results show that biological activities depend on Miyeok manufacture's process. When stimulated macrophages RAW264.7 cells with lipopolysaccharide (LPS), inhibition of NO production in Gijang Sil Miyeok (44.2%) was 9% high comparing with that of Wando Sil Miyeok (35.7%).

Mineral Contents and Physiological Activities of Dried Sea Tangle (Laminaria japonica) Collected from Gijang and Wando in Korea. (기장산과 완도산 건 다시마의 무기성분 및 생리활성 분석)

  • Choi, Jae-Suk;Shin, Su-Hwa;Ha, Yu-Mi;Kim, Yang-Chun;Kim, Tae-Bong;Park, Sun-Mee;Choi, In-Soon;Song, Hyo-Ju;Choi, Young-Ju
    • Journal of Life Science
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    • v.18 no.4
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    • pp.474-481
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    • 2008
  • This research was performed to determine the proximate compositions, mineral contents, alginic acid, antioxidative activities and amino acids of sea tangles collected from Gijang and Wando area. Crude protein and ash contents were higher in Gijang sea tangle, whereas carbohydrate and moisture were higher in Wando in general. Mineral contents of Gijang sea tangle were higher than Wando. Especially, Na and K was the most abundant in both Gijang and Wando sea tangles. Alginic acid content was almost similar in both sea tangles. The major free amino acids were glutamic acid, aspartic acid, alanine, proline and hydroxyproline in both Gijang and Wando sea tangles. Antioxidative activity of methanol extract of sea tangle was measured by using DPPH radical scavenging and SOD-like activity. DPPH radical scavenging and SOD-like activity were about 17% ($40\;{\mu}g/ml$) and 7% ($5\;{\mu}g/ml$) higher, respectively, in Wando sea tangle. When stimulate the macrophages RAW264.7 cells with lipopolysaccharide (LPS), inhibition of NO synthesis of the methanol extract was 11% higher in Wando sea tangle comparing with Gijang samples.

Physiological Activity of Methanol Extracts from Sambucus sieboldiana var. miquelii (Nakai) Hara (지렁쿠나무 메탄올 추출물의 생리활성 연구)

  • Oh, Yu Jin;Cho, Hae Jin;Woo, Hyun Sim;Byeon, Jun-Gi;Kim, Yeong-Su;Kim, Dae Wook
    • Journal of Life Science
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    • v.30 no.11
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    • pp.965-972
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    • 2020
  • Sambucus sieboldiana var. miquelii (Nakai) Hara is distributed in Korea, China, and Japan, and has been used as an anti-rheumatic in folk medicine in oriental countries. The present study aims to investigate the potential use of this species in health functional foods, cosmetics, and food preservatives. Methanol extracts of leaves and branches from this plant were prepared to quantitatively analyze the total phenol and flavonoid contents, and to investigate the antioxidative and enzyme inhibitory activities, and the inhibition of nitric oxide (NO) production activity. The results showed that the total polyphenol and flavonoid contents of the crude extract were 1.52±0.1 mg/g and 1.73±0.1 mg/g, respectively. S. sieboldiana polyphenols exhibited potent scavenging activity shown by 2, 2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and 2, 2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation assay. The crude extract also exhibited significant α-glucosidase and tyrosinase inhibitory activity with IC50 values of 183.5 ㎍/ml and 323.9 ㎍/ ml, respectively. Additionally, the crude extract exhibited strong anti-inflammatory activity determined through the nitric oxide inhibition assay in a dose-dependent manner with an IC50 value of 36.7 ㎍/ml and no cytotoxic effect on the macrophages. Therefore, we demonstrated that the leaves and branches of S. sieboldiana extract possess antioxidant, anti-diabetic, depigmentation potential, and NO production inhibitory activities. According to recent research, S. sieboldiana has great potential as a source of the bioactive compound which could be used as food, cosmetics, and pharmaceutical agents.

Trans-10, cis-12 Conjugated Linoleic Acid Modulates Nuclear Factor-${\kappa}B$ p65 Activity on the Production of Tumor Necrosis Factor-${\alpha}$ in Porcine Peripheral Blood Mononuclear Cells (돼지 말초혈액 단핵구세포에서 trans-10, cis-12 conjugated linoleic acid의 TNF-${\alpha}$ 생산에 대한 nuclear factor-${\kappa}B$ p65 활성 조절 효과)

  • Kim, Young-Beum;Lee, Ill-Woo;Kang, Ji-Houn;Yang, Mban-Pyo
    • Journal of Veterinary Clinics
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    • v.28 no.2
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    • pp.190-195
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    • 2011
  • Nuclear factor ${\kappa}B$ (NF-${\kappa}B$) is a nuclear transcription factor that modulates the expression of inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$. trans-10, cis-12 (t10c12)-conjugated linoleic acid (CLA) participates in the inhibition of TNF-${\alpha}$ production upon lipopolysaccharide (LPS)-stimulation. However, in our previous study, t10c12-CLA enhanced the production of TNF-${\alpha}$ by LPS-unstimulated porcine peripheral blood mononuclear cells (PBMCs) and RAW 264.7 macrophages in vitro. To resolve this apparent contradiction, we hypothesized that the effect of t10c12-CLA on TNF-${\alpha}$ production depends on NF-${\kappa}B$ activation induced by LPS stimulation. To test this hypothesis, we assessed the in vitro effect of t10c12-CLA on TNF-${\alpha}$ production and NF-${\kappa}B$ p65 activity in LPS-stimulated and LPS-unstimulated porcine PBMCs. t10c12-CLA treatment resulted in increased TNF-${\alpha}$ production by LPS-unstimulated PBMCs but decreased TNF-${\alpha}$ production by LPS-stimulated PBMCs. t10c12-CLA increased the degradation of inhibitory ${\kappa}B$ ($I{\kappa}B$)-${\alpha}$ protein and activated NF-${\kappa}B$ p65 in LPS-unstimulated PBMCs, but had the opposite effect in LPS-stimulated PBMCs. Notably, t10c12-CLA enhanced NF-${\kappa}B$ p65 binding activity in LPS-unstimulated PBMCs exposed to caffeic acid phenethyl ester (CAPE), a NF-${\kappa}B$ inhibitor. Conversely, it inhibited NF-${\kappa}B$ p65 binding activity in LPS-stimulated PBMCs exposed to CAPE. These results suggest that t10c12-CLA may have different actions under different physiological conditions, and that its effect may be associated with a change in NF-${\kappa}B$ p65 activity.

Inhibitory Effects of Ethanol Extracts from Pine Buds (Pinus densiflora) on Angiotensin Converting Enzyme, Xanthine Oxidase and Nitric Oxide Synthesis (소나무 새순 에탄올 추출물의 angiotensin converting enzyme, xanthine oxidase 및 nitrix oxide synthase 활성)

  • Cho, Eun-Kyung;Song, Hyo-Ju;Cho, Hea-Eun;Kim, Mi-Hyang;Choi, In-Soon;Choi, Young-Ju
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1629-1636
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    • 2009
  • Pine trees (Pinus densiflora Sieb. et Zacc.) have been used as a traditional health-promoting medicinal food in Korea. This research was performed to determine the antioxidative and antibacterial activities, tyrosinase, nitric oxide synthesis, angiotensin converting enzyme (ACE), and xanthine oxidase inhibition effects of the pine bud ethanol extract (PBE). Antioxidative activities of PBE were measured by using 2,2-diphenyl-1-picryl-hydrazyl (DPPH) free radical scavenging activity and superoxide dismutase-like activity (SODA). DPPH radical scavenging and SOD-like activities of PBE were remarkably increased in a dose-dependent manner, and were about 88.9% and 47.9% at 1 mg/ml and 10 mg/ml, respectively. The xanthine oxidase and angiotensin converting enzyme activities were inhibited about 71.9% and 60.8% at 1 mg/ml and $100{\mu}g/ml$ of PBE, respectively. The tyrosinase inhibitory activities of PBE were slightly increased in a dose-dependent manner. The PBE showed strong antimicrobial activities on Escherichia coli (E. coli) and Vibrio paraheamolyticus. Stimulation of the macrophages RAW264.7 cells with lipopolysaccharide (LPS) resulted in increased production of nitric oxide (NO) in the medium. However, NO synthesis was reduced up to 54% by addition of PBE at $200{\mu}g/ml$. These results revealed that pine buds have a strong antioxidative and anti-inflammatory activity, and exhibit angiotensin converting enzyme and xanthine oxidase inhibitory activities. This suggests that pine buds have the greatest property as a source for natural health products.