Our previous study showed that lungs infected by Pseudomonas, a gram-negative bacteria, produce prostaglandin $D_2$ ($PGD_2$) and prostaglandin $E_2$ ($PGE_2$), the two major prostanoids generated by cyclooxygenase-2 (COX-2), and that the ratio of $PGD_2$ and $PGE_2$ can affect the outcome of the bacterial lung infection. In this study, we sought to uncover the mechanism that determines the ratio of $PGD_2$ and $PGE_2$ produced in lung inflammation. When treated with lipopolysaccharide (LPS), primary alveolar macrophages, extracted from mouse lung, more $PGE_2$ was produced than $PGD_2$, whereas MH-S, a murine alveolar macrophage cell line, produced more $PGD_2$ than $PGE_2$ in a similar experiment. Western blot analyses showed that the kinetics of COX-2 expression in both cell types is similar and epigenetic silencing of COX-2 expression did not affect expressions of lipocalin-PGD synthase (L-PGDS) and PGE synthase (mPGES-1), major enzymes synthesizing $PGD_2$ and $PGE_2$ in inflammation, respectively, indicating no effect of COX-2 on expressions of the two enzymes. Expressions of L-PGDS and mPGES-1 were also similar in both cell types, suggesting no effect of the two key enzymes in determining the ratio of $PGD_2$ and $PGE_2$ in these cells. A single intraperitoneal injection of LPS to C57BL/6 mice induced COX-2 expression and, similar to alveolar macrophages, produced more $PGE_2$ than $PGD_2$ in the lung. These results suggest that the differential expressions of $PGD_2$ and $PGE_2$ in the lung reflect those in alveolar macrophages and may not be directly determined by the enzymes responsible for $PGD_2$ and $PGE_2$ synthesis.
Background: Analysis of cells in bronchoalveolar lavage(BAL) fluid had been used to predict the histologic changes of the bronchioles and alveoli in patients with interstitial lung diseases(ILD). Definitive diagnosis can be a1so made in some cases of ILD, such as histiocytosis. However, there are a few data of the cellular components in BAL fluid in normal Korean individuals and in patients with ILD. In order to evaluate the role of the cellular analysis of BAL fluid in prediction of alveolitis and differential diagnosis among ILDs, we compared the cellular components in BAL fluid from 50 normal individuals and 86 ILD patients. Method: BAL was performed by instillation and retrievement of normal saline with fiberoptic bronchoscopy. The cell number was counted by Hemocytometer. Differential count was done up to 500 cells on slides prepared by Diff-Quik stain and non-specific esterase stain. We compared the recovery rate(RR), cell numbers(CN), and percentages of each cellular components(CP). Results: The results were as follows: 1) There was no difference in RR, CN and CP between the normal smoker group and normal non-smoker group. 2) Total cell numbers recoverd in BAL fluid increased in collagen vascular diseases(CVD), hypersensitivity pneumonitis(HP), idiopathic pulmonary fibrosis(IPF), and miliary tuberculosis(Mil TBC) groups. 3) The percentage of lymphocytes increased in HP, IPF and Mil TBC groups. Macrophage percentages increased in HP, IPF, and Mil TBC groups. Neutrophil percentages were increased in CVD, HP, IPF and Mil TBC groups. Eosinophil percentages were increased in HP, IPF and Mil TBC groups. The numbers of each cells showed same findings as the percentages did. Conclusion: The analysis of cellular components of BAL fluid can predict the presence of alveolitis in many cases of ILDs. However, It was not helpful in differential diagnosis among ILDs.
Da Sol Kim;Kang Mi Kim;Koanhoi Kim;Young Chul Park
Journal of Life Science
/
v.34
no.4
/
pp.271-278
/
2024
Redox factor (Ref)-1, a ubiquitously expressed protein, acts as a modulator of redox-sensitive tran- scription factors and as an endonuclease in the repair pathway of damaged DNA. However, the function of Ref-1 in the differentiation of monocytes into macrophages has not been defined. In this study, we investigated the effects of Ref-1 on the monocyte differentiation process using the human monocytic cell line THP-1. The differentiation agent PMA increased cell adhesion over time and showed a sig- nificant increase in phagocytic function but decreased the intracellular amount of Ref-1. Ref-1 inhibitor E3330 and Ref-1 knockdown using the siRNA technique reduced cell adhesion and the expression of differentiation markers, such as CD14, ICAM-1, and CD11b, by PMA stimulation. This means that the role of Ref-1 is absolutely necessary in the initial process of differentiating THP-1 cells stimulated by PMA. Next, the distribution of Ref-1 was examined in the cytoplasm and nucleus of THP-1 cells stimulated with PMA. Surprisingly, PMA stimulation resulted in the rapid translocation of Ref-1 to the nucleus. To prove that movement of Ref-1 to the nucleus is required for monocyte differentiation, a Ref-1 vector with the nuclear localization sequence (NLS) deleted was used. As a result, overexpression of ∆NLS Ref-1, which restricted movement to the nucleus, suppressed the expression of differentiation markers and notably reduced phagocytic function in PMA-stimulated THP-1 cells. In conclusion, these data suggest that the differentiation of monocytic THP-1 cells requires Ref-1 nuclear translocation during the initial process of biochemical events following stimulation from PMA.
In strawberry farming, most parts of strawberry stems but the fruit have been dumped. Therefore, this study attempted to investigate the antioxidant and anti-inflammatory effects of strawberry stems which are thrown away after farming. For this, strawberry stem extracts were obtained, using hot water and 70% ethanol. First, total polyphenol contents of the hot water and ethanol extract were checked (265.4 ± 0.12 mg TAE/100 g, 503.88 ± 0.2 mg TAE/100 g). For analysis of antioxidant activities, electron donating ability (EDA) and 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity were measured. Both extracts increased in a dose-dependent fashion, and similar effects with vitamin C (control group) were confirmed. In terms of cell viability of the hot water and ethanol extracts of strawberry stems, 'RAW 264.7' was 99% or higher at 500 ㎍/ml. In addition, cell experiments were conducted at 50, 100 and 500 ㎍/ml where cell viability is above 99%. In terms of inhibition of the inflammatory mediator 'nitric oxide (NO)', the hot water and ethanol extracts of strawberry stems were 37.9% and 38.8% respectively, confirming the inhibition of NO production. To check anti-inflammatory activities, protein and mRNA expressions of 'iNOS' and 'COX-2' were measured, using RAW 264.7. Compared to the LPS group, the protein expression of the inflammatory mediators was inhibited in the hot water and ethanol extract-treated groups. The above results confirmed that the hot water and ethanol extracts of strawberry stems are valuable as natural substances with antioxidant and anti-inflammatory activities.
Background and Objective : Haeyeol-tang, composed of Houttuyniae Herba, Lonicerae Flos, Taraxaci Herba, and Scrophulariae Radix, is widely used for alleviating the symptom of various kinds of inflammatory pulmonary disease, including asthma and COPD. We want to know whether Haeyeol-tang has an anti-inflammatory effect by analyzing expression of pro-inflammatory cytokines. Materials and Methods : We differentiated the THP-1 cells into macrophage-like cells by treatment with PMA. Inflammation was induced by treatment with LPS and PMA. We found the safe concentration of Haeyeol-tang by using MTS assay and used PD98059 as a negative control for comparison of anti-inflammatory effect of Haeyeol-tang. Results : The RT-PCR analysis results show that the cell survival rate is over 100% within 1 ng/mL to 1 ug/mL of Haeyeol-tang and begins to decrease under 100% at 10 ug/mL. The gene expression of $IL-1{\beta}$, IL-6, IL-8, IL-10, $TNF-{\alpha}$ and $TGF-{\beta}$ levels were down-regulated when Haeyeol-tang was treated at concentrations between 1 ng/mL an 1 ug/mL on monocyte-derived macrophages. Interestingly, 1 ug/mL Haeyeol-tang-treated samples showed that the transcriptional activities of IL-8, $TNF-{\alpha}$, IL-10 and $TGF-{\beta}$ were more down-regulated than those of PD098059 $(TNF-{\alpha}$ inhibitor). At protein level, the ELISA analysis results showed that there were more remarkable (p<0.001) decreases in expression of $IL-1{\beta}$, IL-6, IL-8 and $TNF-{\alpha}$ on both the 1 ug/mL Haeyeol-tang-treated group and the PD98059-treated group than the LPS-treated group. Conclusion : We conclude that Haeyeol-tang has an anti-inflammatory effect by inhibiting expression of pro-inflammatory cytokines and chemokines at mRNA and protein levels. These results may provide us a promising way to care for general inflammatory diseases as well as inflammatory pulmonary disease, including asthma and COPD, with further clinical study.
Objectives: The object of this study was to observe the in vitro antibacterial effects of five single(Pulsatillae Radix, Patrinae Radix, Sanguisorbae Radix, Sophorae Flos, and Sophorae Radix) aqueous herbal extracts, traditionally used for treating various gynecological diseases including mastitis in Korea, against Staphylococcus aureus. Methods: Antibacterial activities against Staphylococcus aureus of aqueous extracts of Pulsatillae Radix, PatrinaeRadix, Sanguisorbae Radix, Sophorae Flos, and Sophorae Radix were detected using standard agar microdilution methods. In addition, the effects on the bacterial growth curve were also monitored at Minimal Incubation Concentration(MIC) and $MIC{\times}2$ levels. The effects on the intracellular killing and bacterial invasion of individual test materials were also observed using murine macrophage(Raw 264.7) and human mammary gland carcinoma cell(MCF-7). Results: MIC of aqueous extracts of Pulsatillae Radix, Patrinae Radix, Sanguisorbae Radix, Sophorae Flos, and Sophorae Radix against Staphylococcus aureus were detected as $0.215{\pm}0.107$ mg/ml, $0.273{\pm}0.107$ mg/ml, $0.469{\pm}0.297$ mg/ml, $11.850{\pm}8.406$ mg/ml, and $0.664{\pm}0.546$ mg/ml, respectively. MIC of Ciprofloxacin was detected as $0.469{\pm}0.297{\mu}g/ml$ at same conditions. In addition, all five single aqueous herbal extracts were also showed marked dosage-dependent inhibition of bacterial growth. The effects of intracellular killing with Raw 264.7 and inhibition of bacterial invasion with MCF-7 cells were detected, in the order of Sophorae Flos, Pulsatillae Radix, Patrinae Radix, Sanguisorbae Radix and Sophorae Radix aqueous extracts in the present study. Conclusions: The results obtained in this study suggest that all five single aqueous herbal extracts showed antibacterial effects against Staphylococcus aureus and they also showed dosage-dependent inhibitory effects on the bacterial growth. They showed the significant intracellular killing and inhibition of bacterial invasion effects. It means, all five single aqueous herbal extracts may show potent anti-infectious effects against Staphylococcus aureus for mastitis.
Journal of the Korea Society of Computer and Information
/
v.25
no.10
/
pp.115-123
/
2020
In this paper, we propose to evaluate the potential anti-inflammatory properties of root bark extract of Cudrania (C.) tricuspidata on lipopolysaccharide (LPS)-induced inflammation in macrophages of apolipoprotein E (ApoE) knockout (ApoE-/-) mice, murine model of atherosclerosis. Atherosclerosis is a chronic vascular inflammatory disease. C. tricuspidata is a small tree of the Moraceae family and its extract has anti-inflammatory activities. However, its role in the progress of atherosclerosis is not yet clear. To determine anti-inflammatory effects of C. tricuspidata in atherogenesis, we applied LPS in peritoneal macrophages of ApoE-/- mice and measured cell viability by CCK-8 and expression of pro-inflammatory cytokines by qRT-PCR following treatment with root bark extract of C. tricuspidata. Research data was expressed as differences between the cells treated with LPS and root bark extract and the cells treated with LPS alone (control) by a two-tailed non-parametric Mann-Whitney U-test using GraphPad Instat program. No cytotoxic effect was observed when the cells were treated with the extract at concentrations ≤ 100 ㎍/mL. The expression of inflammatory cytokines, including MCP-1, IL-1β, IFN-γ, TNF-α, and IL-6 were inhibited by the extract. These results indicated that the extract has an anti-inflammatory effect and therefore a possible role in the treatment of atherosclerosis.
Lee, Su Hyeon;Jin, Kyong-Suk;Kwon, Hyun Ju;Kim, Byung Woo
Journal of Life Science
/
v.28
no.9
/
pp.1092-1099
/
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.
Jae-Hoon Choi;Hyun-Mi Cho;Heung-Soo Son;Tae-Woong Kim
Journal of the Korean Society of Food Science and Nutrition
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v.23
no.3
/
pp.402-408
/
1994
Human plasma low density lipoprotein (LDL) is the major factor of coronary heart disease.But recent studies suggest the normal LDL can be realdily oxidized by oxygen free radicals and not interact with LDL receptors.Lipoprotei particles consist of lipid and protein, and fatty acids are prone to oxidatioin.The fatty acid compositions of LDL from Koreans was compared with that of Westerners.From the results, the raio of unsaturated fatty acids of korean and Westerner approximately 30 and 70%, respectively.which means Westerners are more labile in the lipid oxidation of LDL than Koreams.Normal LDL was incubated with $CuSO_4$ in PBS to lead for the peroxidation of LDL, and it was tested by the detection of TBARS and free radicals.Then, ascorbate, ${\alpha}-tocopherol$ and hyaluronic acid were found to have effects of antioxidants on LDL oxidation.The amount of free radical increased as the extent of oxidation increased.The time course of free radical formation was similar to TBARS.Therefore, determination of free radical by Luminometer was much more convenient than that of TBARS.
Han, Ik-Hwan;Goo, Sung-Young;Park, Soon-Jung;Hwang, Se-Jin;Kim, Yong-Seok;Yang, Michael Sungwoo;Ahn, Myoung-Hee;Ryu, Jae-Sook
Parasites, Hosts and Diseases
/
v.47
no.3
/
pp.205-212
/
2009
Trichomonas vaginalis commonly causes vaginitis and perhaps cervicitis in women and urethritis in men and women. Macrophages are important immune cells in response to T. vaginalis infection. In this study, we investigated whether human macrophages could be involved in inflammation induced by T. vaginalis. Human monocyte-derived macrophages (HMDM) were co-cultured with T. vaginalis. Live, opsonized-live trichomonads, and T. vaginalis Iysates increased proinflammatory cytokines, such as TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 by HMDM. The involvement of nuclear factor (NF)-${\kappa}B$ signaling pathway in cytokine production induced by T. vaginalis was confirmed by phosphorylation and nuclear translocation of p65 NF-${\kappa}B$. In addition, stimulation with live T. vaginalis induced marked augmentation of nitric oxide (NO) production and expression of inducible NO synthase (iNOS) levels in HMDM. However, trichomonad-induced NF-${\kappa}B$ activation and TNF-${\alpha}$ production in macrophages were significantly inhibited by inhibition of iNOS levels with L-NMMA (NO synthase inhibitor). Moreover, pretreatment with NF-${\kappa}B$ inhibitors (PDTC or Bay11-7082) caused human macrophages to produce less TNF-${\alpha}$. These results suggest that T. vaginalis stimulates human macrophages to produce proinflammatory cytokines, such as IL-1, IL-6, and TNF-${\alpha}$, and NO. In particular, we showed that T. vaginalis induced TNF-${\alpha}$ production in macrophages through NO-dependent activation of NF-${\kappa}B$, which might be closely involved in inflammation caused by T. vaginalis.
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