• Title/Summary/Keyword: NO and cytokines production

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Immunomodulatory effects of six Acetobacter pasteurianus strains in RAW-Blue macrophage

  • Sun Hee Kim;Woo Soo Jeong;So-Young Kim;Soo-Hwan Yeo
    • Food Science and Preservation
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    • v.30 no.1
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    • pp.65-77
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    • 2023
  • In this study, we investigated the immunological properties of six strains of Acetobacter pasteurianus through nuclear factor-kappa B/activator protein-1 (NF-κB/AP-1) transcription factor activation and nitric oxide (NO) and cytokine production in macrophages. We found that the six A. pasteurianus strains had no significant inhibitory effect on the cell viability of RAW-BlueTM cells at the concentration of (25, 50, 100 CFU/macrophage). The production of NO and cytokines (TNF-α, IL-1β, and IL-6) showed different abilities of immune activation for each strain, and it was 0.7 to 0.9 times higher than that of the LPS (100 ng/mL, v/v) positive control and 7 to 8 times superior to that of the negative control group. To explore the underlying mechanism, we evaluated the mRNA expression of pro-inflammatory genes. Consequently, we found that inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 expression including genes expression of cytokines were elevated by the six A. pasteurianus treatment. These results suggested that the six strains of A. pasteurianus have an excellent industrial application value as a functional material for the purpose of enhancing immune function.

Cytokines Stimulate Lung Epithelial Cells to Release Nitric Oxide

  • Robbins, Richard A.;Kwon, O-Jung
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.4
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    • pp.447-454
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    • 1995
  • Cytokine release from alveolar macrophages and subsequent interaction of these cytokines with the bronchial epithelium can induce epithelial cells to release inflammatory mediators. Nitric oxide(NO), a highly reactive gas formed from arginine by nitric oxide synthase(NOS), is known to be involved in inflammation and edema formation, and the inducible form of NOS(iNOS) can be increased by cytokines. In this context, we hypothesized that lung epithelial cells could be stimulated by cytokines released by alveolar macrophages to express iNOS. To test this hypothesis, the murine lung epithelial cell line, LA-4, or the human lung epithelial cell line, A549, were stimulated with culture supernatant fluids from alveolar macrophages. NO production was assessed by evaluating the culture supernatant fluids for nitrite and nitrate, the stable end products of NO. Both murine and human cell culture supernatant fluids demonstrated an increase in nitrite and nitrate which were time- and dose-dependent and attenuated by $TNF{\alpha}$ and IL-$1{\beta}$ antibodies(p<0.05, all comparisons). Consistent with these observations, cytomix a combination of $TNF{\alpha}$, IL-$1{\beta}$, and $\gamma$-interferon, stimulated the lung epithelial cell lines as well as primary cultures of human bronchial epithelial cells to increase their NO production as evidenced by an increase in nitrite and nitrate in their culture supernatant fluids, an increase in the iNOS staining by immunocytochemistry, and an increase in iNOS mRNA by Northern blottin(p<0.05, all comparisons). The cytokine effects on iNOS were all attenuated by dexamethasone. To determine if these in vitro observations are reflected in vivo, exhaled NO was measured and found to be increased in asthmatics not receiving corticosteroids. These data demonstrate that alveolar macrophage derived cytokines increase iNOS expression in lung epithelial cells and that these in vitro observations are mirrored by increased exhaled NO levels in asthmatics. Increased NO in the lung may contribute to edema formation and airway narrowing.

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Immune Enhancing Effect of Boummyunyuck-dan (보음면역단의 면역 증강 효과)

  • 김태균;문석재;원진희;김동웅;이종덕;문구
    • The Journal of Korean Medicine
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    • v.24 no.1
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    • pp.54-64
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    • 2003
  • Objective : To investigate immune enhancing effects of Boummyunyuck-dan (BMD) Methods : In this study I investigated the effect of BMD on cell proliferation and viability. In addition, I investigated production of cytokines (IL-2, IL-4 and $IFN-{\gamma}$), NO, and $TNF-{\alpha}$ in human T-cell leukemia, MOLT-4 cells. The cells were cultured for 24h in the presence or absence of BMD. Result : BMD increased the cell viability by 15% (P<0.05) and enhanced IL-2, IL-4 and $IFN-{\gamma}$ production compared with media control in a dose-dependent manner (P<0.01) at 24h. BMD also increased mRNA and protein expression levels of $IFN-{\gamma}$ in MOLT-4 cells. In addition, I also assessed the effects of BMD on production of NO and $TNF-{\alpha}$ from the peritoneal macrophages because NO and $TNF-{\alpha}$ as a potent macrophage-derived immune reaction regulatory molecule has received increasing attention. However, BMD had no effect on NO and $TNF-{\alpha}$ production in the cells. Conclusion : These data indicate that BMD has some immune-enhancing effect, and that its action may be due to the proliferation and cytokine production of T cells.

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Anti-inflammatory effects of 4,4'-diaminodiphenyl sulfone (dapsone) in lipopolysaccharide-treated spleen cells: selective inhibition of inflammation-related cytokines (지질다당류를 투여한 비장세포에서 4,4'-diaminodiphenyl sulfone (dapsone)의 항염증 효과: 염증 관련 사이토카인의 선택적 억제)

  • Moon, Sun-Young;Joo, Hong-Gu
    • Korean Journal of Veterinary Research
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    • v.55 no.3
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    • pp.199-204
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    • 2015
  • 4,4'-diaminodiphenyl sulfone (dapsone) is a sulfone drug that has antibacterial effects on a variety of bacteria, especially Mycobacterium leprae; thus, it has been used to treat leprosy. Previous studies demonstrated that dapsone inhibits integrin-mediated adherence of neutrophils and production of prostaglandin $E_2$ by polymorphonuclear leukocytes. Hence, dapsone may act in immune cells and regulate cell-mediated inflammation processes. However, its anti-inflammatory effects remain unclear. The present study demonstrated that dapsone modulates the production of inflammation-related cytokines in immune cells. We employed the spleen cells of mice, which are major immune cells, and lipopolysaccharide (LPS) as a causative agent of inflammation for experiments. Dapsone induced a proportional change in splenocyte subsets and the apoptosis of spleen cells. Interestingly, dapsone decreased the production of tumor necrosis factor-alpha and interleukin (IL)-10, but not IL-6, in LPS-treated spleen cells. In other assays, we measured the dapsone-induced production of nitric oxide (NO) and the expression of activation markers of spleen cells. Dapsone decreased NO production in LPS-treated spleen cells. Taken together, our results demonstrate that dapsone has anti-inflammatory effects in immune cells and provide new insight into the potential uses of this agent.

Study on the Effect of Gwanjulbang-5 in Rheumatoid Arthritis (관절 5호방의 류마토이드 관절염 치료효과에 대한 실험적 연구)

  • Choi, Jae-Young;Heo, Dong-Seok;Yoon, Il-Ji;Oh, Min-Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.3
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    • pp.728-735
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    • 2007
  • This study was carried out to know the effect of Gwanjulbang-5(hereinafter refer to GJB-5) to on Rheumatoid Arthritis by using human fibroblast-like synoviocytes(hFLS). We performed several experimetal items : that is cytotoxicity of GJB-5, mRNA expression of pro-imflammatory cytokines in hFLS and production of NO, ROS. The results were obtained as follows : GJB-5 reduced the production of pro-inflammatory cytokines TNF-${\alpha}$, IL-1${\beta}$, IL-6, IL-8 in hFLS, increased the production of TIMP-1. As well as GJB-5 reduced the production of ICAM-1, MMP-3, NOS-II, the production of NO and ROS, and the proliferation of hFLS in proportion to the concentration of GJB-5. In conclusion, these results shows that GJB-5 had immunomodulatory effects in treating rheumatoid arthritis.

Anti-inflammtory effects of the MeOH extract of Petiolus Nelumbinis (LPS로 활성화된 RAW 264.7 cell에서 하경(荷梗)의 염증매개물질 억제효과)

  • Lee, Won-Uk;Jo, Mi-Jeong;Park, Sang-Mi;Jung, Ji-Yun;Kim, Sang-Chan
    • Herbal Formula Science
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    • v.17 no.1
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    • pp.175-185
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    • 2009
  • Petiolus Nelumbinis, branches of lotus leaf or lotus flower is a traditional oriental herbal medicine widely used for treating a superheat or disorder of qi flow. Although there are many clinical results and literature study, it has been rarely conducted to evaluate the immuno-biological activity. The present study was conducted to examine the anti-inflammatory effects of PNM (Petiolus Nelumbinis MeOH extract) in vitro. To determine cytotoxic concentration of PNM, the cells were treated with PNM for 24 h after LPS addition, and the cell viability was tested by MTT assay. Both of dosages (30 and 100 ${\mu}g/ml$) of PNM had no cytotoxicity. In these concentrations, PNM significantly reduced the elevated levels of NO and $PGE_2$ by LPS. These inhibitory effects of PNM were due to the reduced expressions of iNOS and COX-2 protein. TNF-$\alpha$, IL-1$\beta$ and IL-6 are frequently encountered pro-inflammatory cytokines, and LPS plays a key role in inducing to the massive production of these cytokines. Thus, we next determined the levels of these cytokines. Although PNM had no significant inhibitory effect on the production of TNF-$\alpha$, the elevated levels of IL-1$\beta$ and IL-6 by LPS were dose-dependently reduced in PNM-treated groups. These results demonstrate that PNM has anti-inflammatory effects by inhibiting the production of proinflammatory cytokines, NO and $PGE_2$ in LPS-activated macrophage. Moreover, the reduction of NO and $PGE_2$ levels was due to the inhibition of iNOS and COX-2 protein expression by PNM.

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Immunological and Pathological Aspects of Respiratory Tract Infection with Stenotrophomonas maltophilia in BALB/c Mice

  • Zgair, Ayaid Khadem;Chhibber, Sanjay
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1585-1591
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    • 2010
  • A comprehensive study on the production of inflammatory mediators in the lungs of BALB/c mice following infection with Stenotrophomonas maltophilia was conducted. The levels of pro-inflammatory cytokines, tumor necrosis factor alpha (TNF-${\alpha}$), and interleukin-1${\beta}$ (IL-1${\beta}$) were raised in the lungs of infected mice compared with control. The production of anti-inflammatory cytokine IL-10 was slightly delayed. Its peak level was on the $2^{nd}$ day, whereas the peak of pro-inflammatory cytokines was observed on day 1 after intranasal challenge. This was accompanied by a rise in myeloperoxidase (MPO) and malondialdehyde (MDA) on day 1. The increase in MPO levels matched with histopathological observations, as neutrophils infiltration was detected on the first day. Alveolar macrophages (AMs) obtained from infected animals showed a higher rate of uptake and killing when exposed to bacteria in vitro, compared with similar experiments conducted with AMs from normal mice (control). This suggests that AMs were more efficient in cleaning the bacteria. The nitric oxide (NO) production however started early during infection but reached its maximum on the $3^{rd}$ day. No mortality was observed among the infected animals, and infection was resolved by the $5^{th}$ day post infection. No drastic changes in the lung tissue were observed on histopathological examination.

Flower MeOH Extract of Panax Notoginseng Attenuates the Production of Nitric Oxide and Pro-inflammatory Cytokines in LPS-stimulated RA W264.7 Cells (삼칠화(三七花)의 대식세포로부터 LPS에 의해 유도되는 nitric oxide와 전염중성 사이토카인의 생성 억제효과)

  • Joo, Ye-Jin;Jung, Hye-Mi;Seo, Un-Kyo
    • The Journal of Korean Medicine
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    • v.30 no.1
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    • pp.150-162
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    • 2009
  • Objectives: Inflammatory mediators, such as nitric oxide (NO), prostaglandin E2 ($PGE_2$) and pro-inflammatory cytokines, TNF-${\alpha}$ and IL-$1{\beta}$ playa critical role in inflammatory immune response. Therefore, intervention of inflammatory mediator production promises therapeutic benefit for treatment of many chronic inflammatory diseases, such as allergic asthma, rheumatoid arthritis, multiple sclerosis, septic shock and neurodegenerative diseases. In this study, the pharmacological effects of the flower MeOH extract Panax notoginseng (Notoginseng Flos; NF) on inflammation were investigated to address potential therapeutic or toxic effects. Methods: RA W264.7 cells were treated with different concentrations of NF methanol (NF-M) extract in the presence or absence of LPS ($1{\mu}g/m{\ell}$). Results: NF-M extract significantly inhibited LPS-induced production of NO, $PGE_2$ and pro-inflammatory cytokines, TNF-${\alpha}$ and IL-$1{\beta}$ in a dose-dependent manner. In addition, NF-M extract suppressed mRNA expressions and protein levels of iNOS, COX-2 and pro-inflammatory cytokines in LPS-stimulated RA W264.7 cells. Conclusion: These results indicated that NF-M extract inhibits LPS-induced production of inflammatory mediators in macrophages and demonstrated that NF-M extract possesses anti-inflammatory properties in vitro.

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Effect of Brussels Sprouts Extract on Inflammatory Cytokine Inhibition (방울양배추 추출물의 염증성 사이토카인 억제에 미치는 영향)

  • Jae-Hyeok Lee;Jeong-Sook Park
    • Journal of Industrial Convergence
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    • v.21 no.8
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    • pp.69-74
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    • 2023
  • This paper was conducted to examine the effect of Brussels Sprouts Extract on the inhibition of pro-inflammatory cytokines. The inflammatory response is manifested by mediators such as reactive oxygen species and inflammatory cytokines such as TNF-α, IL-1β, and IL-8. Therefore, this paper examined the toxicity to cells using the MTS assay, stimulated RAW264.7 macrophages with lipopolysaccharide (LPS), and stimulated reactive oxygen species such as NO and TNF-α, IL-1β, and IL-8. Inhibition of inflammatory cytokines after treatment with 10 mg/mL, 100 mg/mL, and 1000 mg/mL of Brussels Sprouts Extract was investigated. As a result of the experiment, Brussels Sprouts Extract inhibited NO production, TNF-α and IL-8 in a concentration-dependent manner without cytotoxicity, and showed significant inhibition especially at a concentration of 1000 mg/mL. Brussels Sprouts Extract, which inhibits the production of inflammatory cytokines, suggests the possibility of reducing inflammatory response and controlling inflammation, and can be seen as providing potential as a health functional food or prevention and treatment of inflammation.

Inhibitory Effects of Cheongpochukeo-tang on LPS-induced Inflammation Model (LPS로 유도된 염증모델에 대한 청포축어탕의 억제 효과)

  • Hong, Ka-Kyung;Lee, Soo-Hyung;Jung, Hyun-Tae;Kim, Song-Baek
    • The Journal of Korean Obstetrics and Gynecology
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    • v.34 no.4
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    • pp.12-29
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    • 2021
  • Objective: This study was performed to investigate the inhibitory effect of Cheongpochukeo-tang (CCT) on lipopolysaccharide (LPS)-induced inflammation model. Methods: RAW 264.7 cells were pre-treated with CCT and incubated with LPS (500 ng/ml) after 1 hour. Cell viability was measured by MTT assay to figure out cytotoxicity of CCT. The production of nitric oxide and mRNA expression of pro-inflammatory cytokine were measured. And the activation of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) were examined to figure out molecular mechanisms of CCT's anti-inflammatory effects. In addition, mice survival rate and cytokine levels of serum were observed after treated with CCT. And mice liver tissues were observed and their cytokines levels in liver tissue were measured. Results: CCT did not have cytotoxic effect in RAW 264.7 cells. It inhibited LPS-induced nitric oxide (NO) production, but showed an increase in NO by itself at 2 mg/ml concentration. CCT inhibited mRNA expression of IL-1β, IL-6, TNF-α in a dose dependant and the activaton of MAPKs and NF-κB. In addition, CCT reduced mortality in the LPS-induced mouse model and inhibited production of cytokines in mouse serum and liver tissue. Conclusion: The results suggest that CCT could reduce LPS-induced inflammation by inhibiting MAPKs and NF-κB activaton, NO production, and pro-inflammatory cytokines secretion. Thereby, CCT could be effective medicine for the inflammatory disease.