• Title/Summary/Keyword: Murine macrophages

Search Result 343, Processing Time 0.032 seconds

Inhibitory Effect of Aqueous Extract from Lonicera japonica Flower on LPS-induced Inflammatory Mediators in RAW 264.7 Macrophages. (금은화 수용성 추출물의 LPS 유도 염증매개물 억제 효과)

  • Yun, Young-Gab;Kim, Gyu-Min;Lee, Sung-Jun;Ryu, Seong-Hun;Jang, Seon-Il
    • The Korea Journal of Herbology
    • /
    • v.22 no.3
    • /
    • pp.117-125
    • /
    • 2007
  • Objective : Lonicera japonica (Caprifoliaceae) has long been used for treatment of infectious diseases in oriental countries. The aim of this study was to investigative the effect by which the aqueous extract from flower of L. japonica (LJFAE) inhibited the lipopolysaccharide (LPS)-induced inflammatory mediators in murine macrophages, RAW 264.7 cells Methods : The dried flowers of L. japonica were extracted with distilled water at $100^{\circ}C$ for 7 h. The extract was filtered through 0.45 ${\mu}m$ filter, freeze-dried. The dried extract was dissolved in Hank's balanced salt solution (HBSS) and filtered through 0.22 ${\mu}m$ filter before use. Accumulated nitrite, an oxidative product of nitric oxide (NO), was measured in the culture medium by the Griess reaction. The levels of prostaglandin E2 (PGE2), tumor necrosis factor-$\alpha$ (TNF-$\alpha$), interleukin-1$\beta$ (IL-1$\beta$), and IL-6 production, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were measured by enzyme-linked immunosorbent assay and Western blot analysis. Results: LJFAE (10-400 ${\mu}g$/ml) per se had no cytotoxic effect in unstimulated macrophages, but LJFAE concentration-dependently reduced NO, PGE2, TNF-, IL-l, and IL-6 production and COX-2 activity caused by stimulation of LPS. The levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with LJFAE in a concentration dependent manner. Conclusions : These results suggest that LJFAE suppress the NO and PGE2production in macrophages by inhibiting iNOS and COX-2 expression and these properties may contribute to the anti-inflammatory activity of Lonicera japonica.

  • PDF

Antiinflammatory Effect of Aqueous Extract from Red Pepper on Lipopolysaccharide Induced Inflammatory Responses in Murine Macrophages (홍고추가루 수용성 추출물의 항염증 효과)

  • Kwon, Hyuck-Se;Shin, Hyun-Kyung;Kwon, Sang-O;Yeo, Kyung-Mok;Kim, Sang-Moo;Kim, Bok-Nam;Kim, Jin-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.38 no.10
    • /
    • pp.1289-1294
    • /
    • 2009
  • Inflammation is a pivotal component of a variety of diseases, such as atherosclerosis and tumour progression. Various naturally occurring phytochemicals exhibit antiinflammatory activity and are considered to be potential drug candidates against inflammation-related pathological processes. Red pepper is the most consumed species in Korea. However, the antiinflammatory effects of red pepper have not been characterized. Thus, the present study was designed to evaluate the effects of the aqueous extract from red pepper (RPAE) on lipopolysaccharide (LPS)-induced inflammatory responses in murine macrophages. RPAE demonstrated strong antiinflammatory activity through its ability to reduce nitric oxide and prostaglandin $E_2$ production in the LPS-stimulated mouse macrophage cell, RAW264.7. It also inhibited the production of interleukin-6 (IL-6) on the LPS-stimulated RAW264.7 cells. Further study indicated that LPS-stimulated induction of inducible nitric oxide synthase and cyclooxygenase-2 was significantly inhibited by RPAE exposure (1,000 mg/mL) in RAW264.7 cells. Collectively, these data suggest that the use of RPAE may be a useful therapeutic approach to various inflammatory diseases.

Stimulation of Nitric Oxide Production in RAW 264.7 Macrophages by the Peptides Derived from Silk Fibroin. (실크 피브로인 유래 펩타이드에 의한 RAW 264.7 Macrophage의 Nitric Oxide 생성 촉진)

  • 박금주;현창기
    • Microbiology and Biotechnology Letters
    • /
    • v.30 no.1
    • /
    • pp.39-45
    • /
    • 2002
  • It was found that the peptides originated from the hydrolysates of silk fibroin have in vitro immunostimulating effects in murine macrophage RAW264.7 cells. The stimulation effects on nitric oxide (NO) production resulted from treatments of acid or enzymatic hydrolysates were measured. The silk fibroin preparation isolated from cocoon was most efficiently digested by acid hydrolysis. Even though the sole treatment of acid hydrolysate stimulated the NO production in dose-dependent pattern, a part of its activity was found to be caused by the contaminated endotoxin, LPS. When each endotoxin-free hydrolysates obtained by filtering it through an ultrafiltration membrane of molecular weight (MW) cut-off 10,000 to eliminate LPS was used, the peptic hydrolysate with lowest degree of hydrolysis showed the highest activity. The fractions of peptic hydrolysate with MW ranges of 1,000∼10,000, 500∼1,000 and below 500 also showed a higher MW-higher activity correlation. From the analyses of amino acid composition of each hydrolysate, it was found that the contents of arginine, lysine, alanine and glycine residues affected the activity level of hydrolysate. The results of this study showed a possibility of utilizing fibroin as a source for immunostimulating (chemopreventive) functional peptides.

Immunological properties of the 30 kDa antigen of Toxoplasma gondii (단클론 항체를 이용하여 정제한 톡소포자충 30 kDa 항원의 면역학적 특성)

  • Lee, Yeong-Hwa;No, Tae-Jin;Sin, Dae-Hwan
    • Parasites, Hosts and Diseases
    • /
    • v.35 no.1
    • /
    • pp.55-62
    • /
    • 1997
  • The molecular weight 30 kDa membrane protein of Toxoplusma Sondii (Toxoplasma 30 kDa) apparently conserved in most strains of T. gondii and sera of infected hosts. The present study aimed to elucidate Toxoplasmc 30 kDa as a useful diagnotic antigen for serodiagnisis of toxoplasmosis by ELISA and for induction of protective immunity. Murine spleen cells immunized with the membrane antigen of T. gondii were fused with mouse Sp2/0-Ag 14 myeloma cells. Out of 8 clones selected, five were IgG2b, the others belonged to IgG 1 and IgG2a. The 30 kDa antigen was distributed mainly on the surface membrane of tachyzoites by indirect fluorescence method. Murine peritoneal macrophages which were activated by 30 kDa antigen produced more amounts of NO2 compared with crude antigen-treated group, however there were no significant differences in toxoplamacidal activity between the two groups. Higher specificity of Toxoplosma 30 kDa antigen was recognized for serodiagnosis of toxoplasmosis than the crude antigen. From these results, ToxopLasmo 30 kDa antigen enhances the cytotoxic effect of macrophages as well as a more reliable means for the serodiagnosis of toxoplasmosis by ELISA. Key words: Toxoplosma gondii, 30 kDa antigen (p30), mouse, serodiagnosis, macrophage, cytotoxicity.

  • PDF

Glucosylation of Resveratrol Improves its Immunomodulating Activity and the Viability of Murine Macrophage RAW 264.7 Cells (당화된 레스베라트롤의 대식세포 RAW 264.7세포의 생존능력과 레스베라트롤의 면역제어 활성을 증가)

  • Pandey, Ramesh Prasad;Lee, Jisun;Park, Yong Il;Sohng, Jae Kyung
    • Microbiology and Biotechnology Letters
    • /
    • v.45 no.1
    • /
    • pp.19-26
    • /
    • 2017
  • Effects of resveratrol glucosylation on the immunomodulation properties of resveratrol and on the viability of macrophage cells have been studied by using murine macrophage RAW 264.7 cells. Nitric oxide (NO) and interleukin 6 (IL-6) expression in macrophages in vitro were studied after treatment with different concentrations of (E)-resveratrol, (E)-resveratrol 3-O-${\beta}$-${\small{D}}$-glucoside (R-3-G), or (E)-resveratrol 4'-O-${\beta}$-${\small{D}}$-glucoside (R-4'-G). In vitro viability of RAW 264.7 cells after treatment with the aforementioned three compounds was also studied. As demonstrated by macrophage cell viability assays, two different resveratrol monoglucosides, R-3-G and R-4'-G, exhibited 50-80% reduced cytotoxicity in comparison to (E)-resveratrol in A549 and HepG2 cells. Compared to the resveratrol aglycon, both glucosylated resveratrol derivatives positively modulated NO and IL-6 production in macrophages positively via transcriptionally up-regulating IL-6 and iNOS expression. Conjugation of a glucose moiety on resveratrol was found to enhance the immunomodulating activity of resveratrol and the viability of RAW 264.7 cells.

Activation of Murine Macrophage Cell Line RAW 264.7 by Korean Propolis

  • Han, Shin-Ha;Sung, Ki-Hyun;Yim, Dong-Sool;Lee, Sook-Yeon;Cho, Kyung-Hae;Lee, Chong-Kil;Ha, Nam-Joo;Kim, Kyung-Jae
    • Archives of Pharmacal Research
    • /
    • v.25 no.6
    • /
    • pp.895-902
    • /
    • 2002
  • Monocytes and macrophages playa major role in defense mechanism of the host response to tumor, in part through the secretion of several potent products and macrophage cytokines. Monocytes and tissue macro phages produce at least two groups of protein mediators of inflammation, interleukin 1 (IL-1) and tumor necrosis factor (TNF). Recent studies emphasizes that TNF and IL-1 modulate the inflammatory function of endothelial cells, leukocytes, and fibroblasts. In this study, our work is directed toward studying the in vitro effects of Korean propolis on the ability to induce cellular and secretory responses in murine macrophage cell line, RAW 264.7. It was found that Water Extract of Korean Propolis (WEP) could activate macro phages by producing cytokines. The production of the macrophage cytokines, IL-1 and TNF-$\alpha$, by RAW 264.7 treated with WEP was examined from 2.5 $\mu\textrm{g}$/ml up to 25 $\mu\textrm{g}$/ml with dose dependent manner. Nitric oxide (NO) production was also increased when cells were exposed to combination of LPS and WEP from 2.5 $\mu\textrm{g}$/ml up to 25 $\mu\textrm{g}$/ml. At high dose of WEP (50 to 100 $\mu\textrm{g}$/ml) used to prescribe for anti-inflammatory and analgesic medicine showed inhibition of NO production in LPS-stimulated macrophage. Besides cytokine production, NO release, surface molecule expression and cell morphologic antigen expression were increased in response to the stimulation by WEP. These results suggested WEP may function through macrophage activation.

Combination of Poly-Gamma-Glutamate and Cyclophosphamide Enhanced Antitumor Efficacy Against Tumor Growth and Metastasis in a Murine Melanoma Model

  • Kim, Doo-Jin;Kim, Eun-Jin;Lee, Tae-Young;Won, Ji-Na;Sung, Moon-Hee;Poo, Haryoung
    • Journal of Microbiology and Biotechnology
    • /
    • v.23 no.9
    • /
    • pp.1339-1346
    • /
    • 2013
  • Conventional chemotherapeutic regimens often accompany severe side effects and fail to induce complete regression of chemoresistant or relapsing metastatic cancers. The need for establishing more efficacious anticancer strategies led to the development of a combined modality treatment of chemotherapy in conjunction with immunotherapy or radiotherapy. It has been reported that poly-gamma-glutamate (${\gamma}$-PGA), a natural polymer composed of glutamic acids, increases antitumor activity by activating antigen-presenting cells and natural killer (NK) cells. Here, we investigated the antitumor effect of ${\gamma}$-PGA in combination with cyclophosphamide in a murine melanoma model. Whereas cyclophosphamide alone directly triggered apoptosis of tumor cells in vitro, ${\gamma}$-PGA did not show cytotoxicity in tumor cells. Instead, it activated macrophages, as reflected by the upregulation of surface activation markers and the secretion of proinflammatory factors, such as nitric oxide and tumor necrosis factor ${\alpha}$. When the antitumor effects were examined in a mouse model, combined treatment with cyclophosphamide and ${\gamma}$-PGA markedly suppressed tumor growth and metastasis. Notably, ${\gamma}$-PGA treatment dramatically increased the NK cell population in lung tissues, coinciding with decreased metastasis and increased survival. These data collectively suggest that ${\gamma}$-PGA can act as an immunotherapeutic agent that exhibits a synergistic antitumor effect in combination with conventional chemotherapy.

Evaluation of Boldine Activity against Intracellular Amastigotes of Leishmania amazonensis

  • Salama, Isabel Cristina;Arrais-Lima, Cristina;Arrais-Silva, Wagner Welber
    • Parasites, Hosts and Diseases
    • /
    • v.55 no.3
    • /
    • pp.337-340
    • /
    • 2017
  • Leishmaniasis is a neglected and endemic disease that affects poorest population mainly in developing countries. A lack of adequate and definitive chemotherapeutic agents to fight against this infection has led to the investigation of numerous compounds. The aim of this study was to investigate in vitro activity of boldine against Leishmania amazonensis murine cell infection. Boldine ((S)-2,9-dihydroxy-1,10-dimethoxy-aporphine) is an aporphine alkaloid found abundantly in the leaves/bark of boldo (Peumus boldus Molina), a widely distributed tree native to Chile. The in vitro system consisted of murine macrophage infection with amastigotes of L. amazonensis treated with different concentrations from 50 to $600{\mu}g/ml$ of boldine for 24 hr. Intracellular parasite destruction was assessed by morphological examination and boldine cytotoxicity to macrophages was tested by the MTT viability assay. When cells were treated with $100{\mu}g/ml$ of boldine the reduction of parasite infection was 81% compared with untreated cultures cells. Interestingly, boldine-treatment caused a concentration-dependent decrease of macrophage infection that culminated with 96% of reduction when cells were submitted to $600{\mu}g/ml$ of boldine. Cell cultures exposed to $100{\mu}g/ml$ of boldine and $300{\mu}g/ml$ of $Glucantime^{(R)}$ during 24 hr showed a significant reduction of 50% in parasitized cells compared with cell cultures exposed just to $Glucantime^{(R)}$. The study showed that treatment with boldine produces a better effect than treatment with the reference antimonial drug, glucantime, in L. amazonensis infected macrophage. Our results suggest that boldine is a potentially useful agent for the treatment of leishmaniasis.

Immunomodulatory Effects of a Methanol Extract from Opuntia ficus indica on Murine Splenocytes

  • Ahn, Gin-Nae;Kim, Jin-Hee;Park, Eun-Jin;Lim, Yoon-Kyu;Jeon, You-Jin;Jee, Young-Heun
    • Food Science and Biotechnology
    • /
    • v.18 no.6
    • /
    • pp.1316-1321
    • /
    • 2009
  • Multiple beneficial properties of Opuntia ficus indica (OPF) are well established. In the present study, we have investigated the immunological role of OPF extract (OPFE) on murine splenocytes. OPFE dose- and time-dependently enhanced the proliferation of splenocytes without cytotoxicity. Our results also showed that the number of $CD4^+$ helper T cells and CD45R/$B220^+$ pan B cells increased markedly, but not $CD8^+$ cytotoxic T cells or $CD11b^+$ granulocytes/macrophages. In addition, OPFE significantly decreased the production levels of T helper (Th) 1 type cytokines, interferon (IFN)-$\gamma$, and tumor necrosis factor (TNF)-$\alpha$, although had no significantly differences in those of interleukin (IL)-4, a Th2 type cytokine in concanavalin A (Con A)-stimulated blastogenic cells. Furthermore, OPFE alone strongly increased IL-4 production and decreased TNF-$\alpha$ production even in the absence of Con A. On the basis of these results, this study suggests that OPFE enhances immunity by regulating the pro- and anti-inflammatory response, indicating that this extract exerts a marked immunomodulatory effect, confirming its usefulness as therapy for immune-related diseases.

Effects of Semen jugrandis on the iNOS Expression and Superoxide Formation in the RAW264.7 Cells (호도(胡挑) 추출물이 마우스 대식세포주인 RAW264.7 세포주의 iNOS 발현 및 Superoxide 형성에 미치는 영향)

  • Moon, Goo;Ko, Su-Mi
    • The Journal of Korean Medicine
    • /
    • v.20 no.1 s.37
    • /
    • pp.151-160
    • /
    • 1999
  • Nitric oxide(NO) is synthesized via the oxidation of L-arginine by a family of nitric oxide synthases(NOS), which are either constitutive(cNOS) or inducible(iNOS). The induction of iNOS in tissues can lead to the sustained production of high concentrations of NO which may exert pro-inflammatory effects including vasodilation. edema, cyototoxicity, and its activity can be mediated by various pro-inflammatory cytokine, including interferon ${\gamma}(INF-{\gamma})$. tumor necrosis factor, IL- 1 and IL-6. The enzyme, iNOS, became a new target for pharmacologcal research with the aim to find new substances for the treatment of chronic inflammatory disorders. Murine macrophages produce large amounts of NO when activated with $TFN-{\gamma}$ plus LPS. The murine macrophage-like cell line, RAW 264.7, is a suitable cell model on which to perform vitro studies regarding the iNOS system. Semen jugrandis is a fatty walnut seed found in Korea. The walnut have been used in foik medicine to improve virility, to relieved asthma, and to relieve constipation. Sesquiterpenelactones were isolated from this plant. In the course of screening for NO inhibitory activity from medicnial plants, the aqueous extract of this plant was found to have a significant activity. The result are summarized as followings. 1. The viability of cells incubated in the presence of semen jugrandis increased mare than non incubated cells. 2. Semen jugrandis suppressed the production of NO in tissues dependent on density. 3. Semen jugrandis suppressed the induction of iNOS in tissues dependent on density can lead to reduced production of NO. 4. Semen jugrandis suppressed the production of superoxide in tissue depend on density. According to the above mentioned results, semen jugrandis could be applied production of NO and superoxide can lead to reduction of chronic inflammatary. And as a depence matter come into a virus of microbe and tumor cells.

  • PDF