• Title/Summary/Keyword: RAW 264.7 murine macrophage

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Role of Insulin in the Activation of $NF-{\kappa}B/I{\kappa}B$ Pathway in Macrophage Cells (대식세포주에서 인슐린이 $I{\kappa}B/NF-{\kappa}B$ 경로 활성화에 미치는 영향)

  • Lee, Sang-Min;Jang, Yeon-Sil;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.68 no.3
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    • pp.168-174
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    • 2010
  • Background: Sepsis still has a high mortality rate despite adequate supportive care. Newer therapeutic modalities have been developed but they have generally ended in failure. Recently, insulin was reported to have an anti-inflammatory effect by inhibiting the $I{\kappa}B/NF-{\kappa}B$ pathway, and may have therapeutic potential in sepsis. However, the precise mechanism of the anti-inflammatory effect of insulin is unclear. This study examined the role of insulin in activating $I{\kappa}B/NF-{\kappa}B$ in macrophage. Methods: Raw 264.7 cells, a murine macrophage cell line, were used in this experiment. Western blotting using $I{\kappa}B$ Ab and phosphor-specific $I{\kappa}B$ Ab was performed to evaluate the degradation and phosphorylation of $I{\kappa}B$ cells. For the $I{\kappa}B$ Kinase (IKK) activity, an immune complex kinase assay was performed. The level of interleukin-6 (IL-6) was measured by ELISA to determine the level of proinflammatory cytokine. Results: $I{\kappa}B{\alpha}$ degradation began 30 min after lipopolysaccharide (LPS) treatment. However, an insulin pretreatment suppressed the $I{\kappa}B{\alpha}$ degradation caused by the LPS treatment. The phosphorylation of $I{\kappa}B{\alpha}$ and IKK activity was also inhibited by the insulin pretreatment. Finally, the insulin pretreatment showed a tendency to suppress the induction of IL-6 by LPS. Conclusion: Insulin might have an anti-inflammatory effect though partial inhibition of the $I{\kappa}B/NF{\kappa}B$ pathway in macrophage cell lines.

Fruit Mediated Synthesis of Gold and Silver Nanoparticles Using Lycium chinense and Their Antimicrobial Activity

  • Chokkalingam, Mohan;Huo, Yue;Kang, Jong-Pyo;Mathiyalagan, Ramya;Kim, Yoen-Ju;Yang, Deok-Chun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.94-94
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    • 2018
  • The gold (LC-AuNPs) and silver (LC-AgNPs) nanoparticles were rapidly synthesized by fruit extract of Lycium chinense within 1.15 and 25 min respectively in an eco-friendly way. The synthesized nanoparticles confirmed by relevant surface plasmon resonance peaks for gold and silver nanoparticles at 536 and 480 nm, respectively. FE-TEM results revealed that LC-AuNPs were 20-50 nm and LC-AgNPs were 50-100 nm. The maximum distribution of gold, silver elements and the crystallographic nature of synthesized were confirmed using EDX, elemental mapping and XRD. LC-AgNPs showed inhibitory activity against pathogenic microorganisms such as E. coli and S. aureus, whereas LC-AuNPs did not show inhibitory activity. The LC-AgNps nanoparticles exhibited significant cytotoxicity to human breast cancer MCF7 cell line and less cytotoxicity to non-diseased RAW264.7 (murine macrophage) cells whereas LC-AuNps showed minimal toxicity to both cell lines. In-depth research on this rapid, facile and greenery nanoparticles may play a potential role in biomedical applications.

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Anti-oxidative Activity of Five Plant Extracts including Apios fortune, Colubrina arborescens, Croton caudatus, Osmanthus matsumuranus and Schima noronhae (Apios fortunei, Colubrina arborescens, Croton caudatus, Osmanthus matsumuranus, 그리고 Schima noronhae를 포함하는 5종 식물 추출물의 항산화 활성)

  • Lee, Su Hyeon;Jin, Kyong-Suk;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1092-1099
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    • 2018
  • This study was orchestrated with the purpose of uncovering new nutraceutical resources possessing biological activities in the plant kingdom. To fulfill our objective, we analyzed several plant extracts and selected five species possessing powerful anti-oxidative activity. The anti-oxidative effect of these five plants, Apios fortunei Maxim., Colubrina arborescens Sarg., Croton caudatus Geiseler, Osmanthus matsumuranus Hayata and Schima noronhae Reinw. ethanol extracts were then evaluated by using in vitro assay, cell model system, and Western blot analysis of target proteins. As the results, all of them possessed the potent scavenging activity against 1,1-diphenyl-2-picryl hydrazyl (DPPH), similar with that of ascorbic acid, used as a common positive control. Moreover, they strongly inhibited hydrogen peroxide ($H_2O_2$)-induced reactive oxygen species (ROS), in a dose-dependent manner, in RAW 264.7 murine macrophage cells. Furthermore, they induced the protein expression of an anti-oxidative enzyme, heme oxygenase 1 (HO-1), and its upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2). Taken together, these results indicate that these five plants possess potent anti-oxidative activity and thus appear to be useful sources as potential anti-oxidant agents. Therefore, they might be utilized as promising materials in the field of nutraceuticals.

Immunostimulatory and Bone Health-Promoting Activities of Litsea japonica Fruit Extract (까마귀쪽나무 열매 추출물의 면역증가 및 뼈 건강 효과)

  • Park, Sung-Jin;Kang, Jun-Chul;Lee, Da-young;Cho, Ju-Hyun;Yoon, Min-Ho
    • Journal of Food Hygiene and Safety
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    • v.35 no.3
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    • pp.284-289
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    • 2020
  • This study was performed with hot water extract (LJF-W) and 70% ethanol extract (LJF-70E) of Litsea japonica fruit to investigate the immunostimulatory activity and bone health-promoting effect of L. japonica fruit. The production of pro-inflammatory mediator (nitric oxide) and pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) in murine macrophage RAW264.7 cells were estimated to examine the immunostimulatory activity of the fruit extracts. The immunostimulatory activity of LJF-W was higher than that of positive control (geinsenosides). However, there was no effect in LJF-70E. Futhermore, both LJF-W and LJF-70E appeared to stimulate the proliferation of MC3T3-E1 cells, proving the effect as a bone health agent. From this result it could be presumed that L. japonica fruit extracts not only stimulate the immune system, but also the ability to promote bone health.

Composition of Organic Acids and Physiological Functionality of Commercial Makgeolli (시중 유통 막걸리의 유기산 조성과 생리활성)

  • Lee, Sang-Jin;Kim, Ji-Hye;Jung, Yong-Woo;Park, Sun-Young;Shin, Woo-Chang;Park, Cheon-Seok;Hong, Sung-Youl;Kim, Gye-Won
    • Korean Journal of Food Science and Technology
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    • v.43 no.2
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    • pp.206-212
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    • 2011
  • Makgeolli is Korean traditional alcoholic beverage that has historically been brewed. In this study, we analyzed the profile of organic acids in makgeolli and also evaluated its physiological characteristics. Makgeolli contained excess lactic acid, which is produced by lactic acid bacteria (LAB). Anti-obesity effects of makgeolli were investigated in 3T3-L1 preadipocytes. Compared to the negative control, makgeolli inhibited the differentiation of preadipocyte as quantified by Oil red O dye. In particular, $100{\mu}g/mL$ makgeolli reduced 40 to 70% of differentiation. To evaluate the anti-angiogenic and anti-inflammatory effects of makgeolli, we performed chorioallantoic membrane assay and measured nitric oxide production from lipopolysaccharide-induced RAW264.7 cells. Most makgeolli interrupted the formation of neo-vasculature and significantly inhibited NO production in a dose-dependent manner. Taken together, these findings suggest that commercial makgeolli has inhibitory activities against adipogenesis, neo-vascularization, and inflammation, and also they are influenced by second metabolites from nuruk microflora containing fungi and LAB.

Anti-inflammatory effect of ethanol extract from Ecklomia cava on gram-positive bacteria(Peptidoglycan)-induced macrophages (그람-양성균(Peptidoglycan)으로 유도한 대식세포에서 감태 에탄올 추출물의 항염증 효과)

  • Kang, Ok-Hwa;Kim, Sung-Bae;Keum, Joon-Ho;Mun, Su-Hyun;Kim, Yong-Sik;An, Byung-Kwan;An, Hyeon-Jin;Kwon, Dong-Yeul
    • Herbal Formula Science
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    • v.19 no.1
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    • pp.195-205
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    • 2011
  • Objectives : Ecklonia cava is brown alga(Laminariaceae) which grows is sea, it has antioxidant, diarrhea and anticoagulant effect. In this study, the effect of ethanol extract of Ecklonia cava (EC) on peptidoglycan(PGN) -induced NO production in RAW 264.7 cells was investigated. Methods : In the present study, IL-6 production was measured using the enzyme-linked immunosorbent assay(ELISA), prostaglandin $\E_2$($\PGE_2$) production was measured using the EIA kit, and inducible NO synthase(iNOS), cyclooxygenase-2(COX-2), and mitogen-activated protein kinase(MAPK) activation, as determined by western blot analysis and reverse transcription -polymerase chain reaction(RT-PCR). Results : EC inhibited PGN-induced NO and IL-6 production. Consistent with these observations, the protein expression of iNOS and COX-2 were inhibited by EC. Moreover, EC suppressed the phosphorylation of extracellular signal-regulated kinase(ERK) 1/2 in PGN-induced RAW 264.7. Conclusions : These results suggest that EC has inhibitory effects on PGN-induced $\PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the MAPKs phosphorylation.

Anti-colorectal Cancer and Anti-oxidant Activities of Rubiae radix Ethanol Extract in vitro (천초근 에탄올 추출물의 항산화 효능 및 대장암 세포 억제 효과)

  • Nho, Jong Hyun;Sim, Mi Ok;Jung, Ho Kyung;Lee, Mu Jin;Jang, Ji Hun;Jung, Da Eun;Sung, Tae Kyoung;An, Byeong Kwan;Cho, Hyun Woo
    • Korean Journal of Plant Resources
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    • v.31 no.2
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    • pp.102-108
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    • 2018
  • Rubiae radix is root of Runia akane Nakai, it has been used to hemostasis and blood stasis in Korean and China. This study investigated that anti-oxidant and anti-colorectal cancer effect of ERA (ethanol extract of Rubiae radix) and WRA (water extract of Rubiae radix) using RAW 264.7 (murine macrophage from blood) and HCT-116 cells (human colorectal cancer cell line). ERA contained polyphenol ($45.77{\pm}2.03mg/g$) and flavonoid ($22.82{\pm}1.33mg/g$). $500{\mu}M$ $H_2O_2$-induced ROS generation was diminished by $500{\mu}g/m{\ell}$ ERA treatment in RAW 264.7 cells, but not WRA (125, 250, and $500{\mu}g/m{\ell}$). Moreover, caspase-3 activity and DNA fragmentation increased by $500{\mu}g/m{\ell}$ ERA treatment during apoptotic cell death in HCT-116. Results demonstrated that anti-cancer effect of ERA against human colorectal cancer cells is mediated apoptotic cell death and DNA fragmentation through caspase-3 activation. However, further study is required to what active ingredient of ERA are important for anti-oxidant and anti-colorectal cancer effect in vivo.

Antioxidant, anti-inflammatory, and antimicrobial activity of hesperetin and its cyclodextrin inclusion complexes (헤스페레틴(Hesperetin)과 사이클로덱스트린(Cyclodextrin) 포접 복합체의 항산화, 항염증, 항균 활성 )

  • Sung-Sook Choi;Kyung-Ae Lee
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.5
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    • pp.988-1000
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    • 2023
  • Hesperetin(HT) is a potent antioxidant flavonoid aglycone derived from hesperidin(HD). The antioxidant, anti-inflammatory, and antimicrobial activities of HT and its cyclodextrin(CD) inclusion complexes were compared in vitro. HT was prepared by enzymatic hydrolysis of HD, and HT/CD complexes were prepared using 𝛽-cyclodextrin(𝛽-CD) and hydroxypropyl-𝛽-cyclodextrin(HP-𝛽-CD) by solvent co-evaporation method. The solubility of the HT/HP-𝛽-CD inclusion complex increased 93.5-fold compared to HT, and the solubility of HT/𝛽-CD increased 22.5-fold. The HT/HP-𝛽-CD inclusion complex showed a similar effect as HT on radical scavenging activity in antioxidant assays, whereas the HT/𝛽-CD inclusion complex showed slightly lower activity than HT. Cytotoxicity was low in the following order; HT/HP-𝛽-CD, HT/𝛽-CD, and HT in murine macrophage RAW264.7 cells. Treatment with HT and HT/CD inclusion complexes reduced the levels of inflammatory mediators such as nitric oxide(NO), tumor necrosis factor-𝛼(TNF-𝛼) and interleukin-6(IL-6) in the cells. HT and HT/HP-𝛽-CD inclusion complex were more effective than HT/𝛽-CD inclusion complex at relatively low concentrations. Inhibitory effects were tested on skin-pathogenic bacteria, Staphylococcus aureus and Pseudomonas aeruginosa, and they showed an antimicrobial effect on S. aureus in the order of HT = HT/HP-𝛽-CD > HT/𝛽-CD, but they did not show any significant inhibitory effect on P. aeruginosa. In conclusion, HT, the aglycone form of HD, and its CD inclusion complexes showed various biological activities. HT/HP-𝛽-CD inclusion complex, which is the highly soluble form of HT, showed relatively higher activity compared to HT/𝛽-CD inclusion complex.

Anti-inflammatory Activity of Sargassum micracanthum Water Extract (잔가시 물 추출물의 항염증 효과)

  • Jeong, Da Hyun;Kang, Bo Kyeong;Kim, Koth Bong Woo Ri;Kim, Min Ji;Ahn, Dong Hyun
    • Journal of Applied Biological Chemistry
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    • v.57 no.3
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    • pp.227-234
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    • 2014
  • The anti-inflammatory effect of Sargassum micracanthum water extract (SMWE) was investigated using lipopolysaccharide (LPS)-induced inflammatory response in this study. The murine macrophage cell line RAW 264.7 cells were used and MTT assay was performed to measure the cell proliferation ability. The secretion of nitric oxide (NO), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-6 (IL-6), and IL-$1{\beta}$ was measured in LPS-induced RAW 264.7 cells by ELISA. The expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and nuclear transcription factor-kappa B p65 protein was studied by immunoblotting. The Balb/c mice were used for an acute toxicity test, and imprinting control region mice were purchased to evaluate a croton oil-induced ear edema. As a result, there was no cytotoxicity in the macrophage proliferation treated with SMWE compared to the control. NO levels decreased with increasing concentration of SMWE and were inhibited over 50%. Moreover, the secretion of IL-6, TNF-${\alpha}$, and IL-$1{\beta}$ was suppressed in a dose-dependent manner, especially, IL-$1{\beta}$ inhibition activity was over 50% at 50 ${mu}g$/mL. The formation of ear edema of mice was reduced at the highest dose tested compared to that in the control. Moreover, in acute toxicity test, no moralities occurred in mice administered 5,000 mg/kg body weight of SMWE over 2 weeks observation period. These results suggested that SMWE may have significant effects on inflammatory factors and be potential anti-inflammatory therapeutic materials.

Effects of Scutellaria radix Extract on Osteoblast Differentiation and Osteoclast Formation (황금 추출물이 조골세포와 파골세포의 활성에 미치는 영향)

  • Shin, Jeong-Min;Park, Chan-Kyung;Shin, Eun-Ju;Jo, Tae-Hyung;Hwang, In-Kyeong
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.674-679
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    • 2008
  • Scutellaria radix (SR) has been utilized as a traditional medicine for a variety of diseases including Rheumatoid arthritis and its major flavonoids - baicalein, baicalin, and wogonin - have been reported to exert beneficial health effects, including anti-bacterial, anti-viral, anti-inflammatory, and free-radical scavenging. However, the mechanisms underlying this effect remain poorly understood. The principal objective of this study was to determine the effect of SR on osteoblast and osteoclast cells. SR extract was prepared using 70% ethanol solvent. Osteoblastic MC3T3-E1 cells and osteoclast precursor Raw 264.7 macrophage cells were utilized. SR extract increased MC3T3-E1 cell proliferation and stimulated alkaline phosphatase activity dose-dependently, 152.0% of the control at concentration $1{\mu}g/mL$. Additionally, SR extract ($1{\mu}g/mL$) stimulated Bone nodule formation activity in MC3T3-E1 cells, approximately 223.3% of the control, 20 days after the exposure. In addition, SR extract significantly reduced the number of tartrate-resistant acid phosphatase-positive (TRAP+) multinucleated cells from Raw 264.7 cells. In conclusion, SR extract stimulates the proliferation and bioactivities of boneforming osteoblasts, and inhibits the activities of bone-resorbing osteoclasts to a certain degree.