The effect of hyperbaric oxygen on the function of peritoneal macrophages in septic mice / 中华急诊医学杂志

Objective To investigate the changes of peritoneal macrophage function in sepsis mice after hyperbaric oxygen (HO) therapy.Methods One hundred C57BL/6 mice were randomly(random number) divided into four groups in equal number (n=25):control group (normal saline),sepsis group,hyperbaric oxygen group,sepsis+ hyperbaric oxygen group;the sepstic mice were treated by intra-peritoneal injection with zymosan;The mice of hyperbaric oxygen group and sepsis+ hyperbaric oxygen group received hyperbaric oxygen therapy (2 atm,2 h,100％ O2).The number of surviving mice was obsevred at 72 h after the intra-peritoneal injection with zymosan;The percentage of M1 macrophage was detected by flow cytometry.The expression of mRNA of inducible nitric oxide synthase (iNOS) in peritoneal macrophage was detected with RT-PCR.The levels of IL-12 and IL-10 in peripheral blood were measured by ELISA.Results Compared with sepsis group,the number of surviving mice of sepsis+ hyperbaric oxygen group was significantly higher (9 vs.16,P＜0.01).The percentages of M1 macrophage in sepsis+ hyperbaric oxygen group was significantly lower than that in sepsis group [(4.75+0.14)％ vs.(2.25±0.16)％,F=803.438,P＜0.01].The expression of mRNA of inducible nitric oxide synthase(iNOS) in sepsis+ hyperbaric oxygen group were significantly lower than that in sepsis group [(39.62+1.74) vs.(48.32±3.34),F=31.992,P＜0.01].The level of IL-12 in peripheral blood in sepsis+ hyperbaric oxygen group was significantly lower than that in sepsis group [(242.62±19.10) pg/mL vs.(159.51±6.35)pg/mL,F=102.282,P＜0.01].The level of IL-10 was significantly higher than that in sepsis group [(521.26±6.3) pg/mL vs.(188.83±8.53) pg/mL,F=5 896.006,P＜0.01].Conclusions Hyperbaric oxygen therapy had effective effect on increasing the number of surviving mice of sepsis group,reducing the level of IL-12 in peripheral blood and rising the level of IL-10 in peripheral blood.By inhibiting the expression of M1 macrophage and iNOS,hyperbaric oxygen could reduce the excessive inflammatory response and play a role in the protection of mice.

Polymyxin B as an inhibitor of lipopolysaccharides contamination of herb crude polysaccharides in mononuclear cells / 中国天然药物

Lipopolysaccharides (LPS) contamination in herbal crude polysaccharides is inevitable. The present study was performed to explore the effect of polymyxin B on abolishing the influence of LPS contamination in mononuclear cells. LPS was pretreated with polymyxin B sulfate (PB) at different concentrations for 1, 5 or 24 h, and then used to stimulate RAW264.7 and mouse peritoneal macrophages (MPMs). The nitric oxide (NO) and tumor necrosis factor-α (TNF-α) in cell culture supernatant, as the indications of cell response, were assayed. Bupleurum chinensis polysaccharides (BCPs) with trace amount contamination of LPS was treated with PB. 30 μg·mL of PB, treating LPS (10 and 1 000 ng·mL in stimulating RAW264.7 and MPMs respectively) at 37 °C for 24 h, successfully abolished the stimulating effect of LPS on the cells. When the cells were stimulated with LPS, BCPs further promoted NO production. However, pretreated with PB, BCPs showed a suppression of NO production in MPMs and no change in RAW264.7. In the in vitro experiments, LPS contamination in polysaccharide might bring a great interference in assessing the activity of drug. Pretreatment with PB (30 μg·mL) at 37 °C for 24 h was sufficient to abolish the effects of LPS contamination (10 and 1 000 ng·mL).

Mechanisms for the regulatory effect of prostaglandin E2/ prostaglandin E receptor 4 on high mobility group box-Ⅰ protein in lipopolysaccharide-induced sepsis in mouse peritoneal macrophage / 中南大学学报(医学版)

Objective:To explore the effect of prostaglandin E2 (PGE2) on the expression of high mobility group box-1 protein (HMGB1) in peritoneal macrophages of septic mice and its possible mechanisms.Methods:Ihe mouse peritoneal macrophages were isolated and cultured by conventional methods.The model of inflammation was established by using lipopolysaccharide (LPS) to incubate with mouse peritoneal macrophages.The PGE2,prostaglandin E receptor (EP) 4 agonist,EP4 RNAi,and DN.CREB inhibitory plasmid were used to interfere with the LPS-treated mouse peritoneal macrophage.The levels of HMGB 1 was determined by Western blot.Results:Compared with LPS alone treatment,the expression of HMGB 1 in peritoneal macrophages was increased obviously after 24 h by treatment with PGE2 and LPS,and it was also increased after the combined treatment of EP4 receptor agonist with LPS for 24 h (both P0.05);compared with LPS alone treatment,the combined treatment of EP4 receptor agonist with LPS for 24 h could up-regulate the phosphorylation of epidermal growth factor receptor (EGFR) and protein kinase B (Akt) thr308 (P<0.05),which were blocked by EGFR inhibitor.Once Akt specific inhibitor was used before EP4 and LPS treatment,the expression of HMGB1 was declined (P<0.05).Conclusion:PGE2 can up-regulate the expression of HMGB1 in sepsis of peritoneal macrophages through EP4 receptor,which may be related to the activation of EGFR/PI3K/Akt signaling pathway.

Metformin enhances ATP-stimulated inflammasomeactivation in LPS-primed peritoneal macrophages / 中国药理学通报

Aim To explore the influence of metformin(a first-line drug for type 2 diabetes) on ATP-induced inflammasome activation and the release of interleukin-1β(IL-1β) by LPS-activated peritoneal macrophages, a commonly-used inflammatory cell model.Methods Peritoneal macrophages were elicited by intraperitoneal injection of 30 g·L-1 thioglycollate into C57BL/6 mice.Inflammasome was activated and cell pyroptosis was induced by LPS plus ATP treatment, and the pyroptotic cells were calculated after propidium iodide(PI) staining.The protein levels of IL-1β and caspase-1 expressed in the cells and released from them into the supernatant were evaluated by Western blot.Immunofluorescent microscopy was recruited to detect the subcellular distribution and fluorescent intensity of the purinergic P2X7 receptor(P2X7R).Results Metformin per se did not induce pyroptosis in LPS-activated peritoneal macrophages, but it significantly and dose-dependently increased cell pyroptosis induced by ATP treatment.At protein levels, maturated IL-1β(17 ku) could not be released from the cells upon single LPS or LPS plus metformin stimulation;but after ATP was added, maturated IL-1β was released into the supernatants of the cells.Moreover, metformin dose-dependently increased the protein levels of both maturated IL-1β and active caspase-1 released by the LPS-activated peritoneal macrophages upon ATP stimulation.Conclusion Metformin intensifies the activation of inflammasome and increases the release of active caspase-1 and maturated IL-1β upon ATP stimulation in the LPS-activated peritoneal macrophages, which should promote inflammatory responses.

Therapeutic switching: from antidermatophytic essential oils to new leishmanicidal products

This study examined whether the antidermatophytic activity of essential oils (EOs) can be used as an indicator for the discovery of active natural products against Leishmania amazonensis. The aerial parts of seven plants were hydrodistilled. Using broth microdilution techniques, the obtained EOs were tested against three strains of dermatophytes (Trichophyton mentagrophytes, Microsporum gypseum and Microsporum canis). To compare the EOs antifungal and antiparasitic effects, the EOs activities against axenic amastigotes of L. amazonensis were concurrently evaluated. For the most promising EOs, their antileishmanial activities against parasites infecting peritoneal macrophages of BALB/c mice were measured. The most interesting antifungal candidates were the EOs from Cymbopogon citratus, Otacanthus azureus and Protium heptaphyllum, whereas O. azureus, Piper hispidum and P. heptaphyllum EOs exhibited the lowest 50% inhibitory concentration (IC50) values against axenic amastigotes, thus revealing a certain correspondence between both activities. The P. hispidum EO was identified as the most promising product in the results from the infected macrophages model (IC50: 4.7 µg/mL, safety index: 8). The most abundant compounds found in this EO were sesquiterpenes, notably curzerene and furanodiene. Eventually, the evaluation of the antidermatophytic activity of EOs appears to be an efficient method for identifying new potential drugs for the treatment of L. amazonensis.

Effects of Mcl-1 silencing on apoptosis of mouse peritoneal macrophages infected with different virulence of Mycobacterium tuberculosis / 中国病理生理杂志

AIM:To investigate the effect of inhibiting Mcl-1 gene expression on apoptosis of mouse peritoneal macrophages infected with different virulence of Mycobacterium tuberculosis using a technique of RNA interference .METH-ODS:The BALB/c mice were infected with prepared bacterium of the virulence strains of Xinjiang , H37Rv, H37Ra and BCG.Mcl-1-shRNA was applied to the mouse model of infection , and the control groups were set up .On 1 d, 3 d, 5 d and 7 d, the mouse peritoneal macrophages were collected .The expression of Mcl-1 at mRNA and protein levels was deter-mined by real-time PCR and Western blot .The apoptotic rate of peritoneal macrophages was analyzed by flow cytometry . RESULTS:The expression of Mcl-1 at mRNA and protein levels was up-regulated in the peritoneal macrophages from the mice infected with different virulence of Mycobacterium tuberculosis, and the cells from the mice infected with virulence strains of Xinjiang and H37Rv expressed higher level of Mcl-1 than the uninfected control cells (P<0.05).The expres-sion of Mcl-1 at mRNA and protein levels was reduced by RNA interference as compared with control group ( P<0.05 ) . Inhibition of Mcl-1 expression induced apoptosis of peritoneal macrophages in the mice .CONCLUSION: The Mcl-1 ex-pression at mRNA and protein levels in mouse peritoneal macrophages infected with different virulence of Mycobacterium tu-berculosis was effectively suppressed by Mcl-1-shRNA, which can induce macrophage apoptosis .

Effects of Chitosan on the Secretion of Cytokines and Expression of Inducible Nitric Oxide Synthase mRNA in Peritoneal Macrophages of Broiler Chicken

An in vitro experiment was conducted to study the effects of chitosan on the secretion of cytokines and expression of inducible nitric oxide synthase mRNA in peritoneal macrophages of broiler chicken. In the experiment, peritoneal macrophages were incubated for 24 h in culture medium supplemented with 0 (control), 40, 80, 160 and 320 µg/mL chitosan. The results showed that chitosan tended to increase quadratically the levels of interleukin-1 (P = 0.093) and interleukin-2 (P = 0.106) in the culture fluid of peritoneal macrophage. Chitosan also significantly enhanced inducible nitric oxide synthase mRNA expression of peritoneal macrophage in a quadratic dose-dependent manner (P < 0.05) and tended to promote quadratically the secretion of nitric oxide (P = 0.053) and inducible nitric oxide synthase (P = 0.157) in peritoneal macrophages. This result implied that one of the mechanisms by which chitosan modulated immune functions in chickens might be chitosan activating expression of inducible nitric oxide synthase and then improving the secretion of nitric oxide.

Recombinant TAT-CD137 Ligand Cytoplasmic Domain Fusion Protein Induces the Production of IL-6 and TNF-alpha in Peritoneal Macrophages

BACKGROUND: The ligand for CD137 (CD137L; also called 4-1BBL) is mainly expressed on activated APCs such as dendritic cells, B cells and macrophages. Even though CD137L functions as a trigger of the CD137 signaling pathway for T cell activation and expansion, engagement of CD137L can deliver a signal leading to the production of proinflammatory cytokines in macrophages. METHODS: We generated cell-permeable TAT-CD137L cytoplasmic domain fusion protein (TAT-CD137Lct) and examined its ability to initiate the CD137L reverse signaling pathway. RESULTS: Treatment of TAT-CD137Lct induced the production of high levels of IL-6 and TNF-alpha mRNAs and proteins in peritoneal macrophages. TAT-CD137Lct increased phosphorylation of Erk, p38 MAPK and Jnk, and activated transcription factors C/EBP and CREB. However, TAT-CD137Lct did not visibly affect the degradation of the inhibitor of NF-kB (IkBalpha). We further demonstrated that JNK activation was required for TAT-CD137Lct-induced production of TNF-alpha, while activation of Erk and p38 MAPK were involved in IL-6 and TNF-alpha production. CONCLUSION: Our results suggest that TAT-CD137Lct is an effective activator for the CD137L reverse signaling pathway.

Inhibition of Emodin on LPS-induced Nitric Oxide Generation by Suppressing PLC-γ Phosphorylation in Rat Peritoneal Macrophages / 中草药·英文版

Objective To investigate the inhibitory mechanism of emodin on lipopolysaccharide(LPS)-induced nitric oxide(NO)generation in rat peritoneal macrophages.Methods NO production and iNOS expression were measured through nitrite assay and Western blotting assay,respectively.NF-kB activity and nuclei P65 expression were estimated by dual-luciferase and Western blotting assay,respectively.Intracellular free Ca2+([Ca2+]i)was detected using the ratiometric fluorescent calcium indicator dye,Fura-2,and a microspectrofluorometer.PLC-γphosporylation was analyzed by Western blotting assay.Results First,emodin was found playing active roles in suppressing LPS-induced NF-kB activation in rat peritoneal macrophages.Second,emodin down-regulated transient[Ca2*]i and could increase in NF-kB upstream signal.Finally,emodin suppressed phosphorylation of PLC-γ by LPS stimulation in the upstream of[Ca2+]i.Conclusion Suppression of PLC-γ phosphorylation is involved in emodin inhibiting NO generation by LPS stimulation in rat peritoneal macrophages.

Staphylococcal catalase regulates its virulence and induces arthritis in catalase deficient mice.

To figure out whether in vivo expression of Staphylococcal catalase could correlate with the virulence and pathogenicity of the bacteria in the catalase deficient Swiss albino mice. 3 Amino 1, 2, 4 triazole (ATZ) (2 mg/g body wt) treated catalase deficient mice were infected with virulent S. aureus and bacterial burden, antioxidant enzyme levels were estimated after 3, 5 and 10 days of infection. Arthritic scores and levels of serum uric acid in mice were also determined. ATZ treatment was found to have slowed down the clearance of bacteria from blood and their rapid elimination from spleen. Increased tissue catalase activities in the spleen and liver of ATZ pre-treated mice even after 5 days of infection suggested its bacterial origin. It was further verified by zymographic analysis. Increased swelling of joints was observed after 5 days of infection. Uric acid level was found lesser in ATZ treated mice. ATZ treatment slowed the bacterial passage from blood with a lower tissue anti-oxidant enzymes leading to induction of joint inflammation.

Efecto inmunosupresor de Trypanosoma lewisi (Kinetoplastidae) sobre la multiplicación de Toxoplasma gondii (Sarcocystidae) en macrófagos alveolares y peritoneales de rata blanca

The immunosuppressant effect of T. lewisi (Kinetoplastidae) infection on the multiplication of Toxoplasma gondii (Sarcocystidae) on alveolar and peritoneal macrophages of the white rat. The immunosuppressant effect of T. lewisi infection on the multiplication of T. gondii was compared in peritoneal (MP) and alveolar macrophages (MA) of white rat. Two animal groups were infected with T. lewisi and sacrificed after four days and seven days post infection. A group without infection was maintained as a control. The number of intracellular parasites (tachyzoites) (IT) was counted by light microscopy, calculating the rate infection rate per 100 total cells (TC) and per infected cells (IC) for each group of phagocyte cells. The relation quotient IT, TC or IC multiplied percent, provided a statistical ratio (RE) of the relative number of parasites in both cellular types for each time interval. MA as well as MP obtained after 4 days showed a significant increase in the multiplication of T. gondii with respect to the control. Unlike the MP (which had an increase in the multiplication of T. gondii the fourth day of infection with T. lewisi diminishing towards the seventh day), the MA had an increase in the multiplication of the parasite from the fourth to the seventh day. This difference can be related to the route of infection used for the experiments, that affect the MP directly with a greater effect in comparison with the MA of the lungs. Lung compartment will be affected later, when the infection becomes systemic between the fourth and sixth day of infection. The immunity against T. gondii is similar between both phagocytes, but the time of infection and the compartment where the cells are located, makes the difference in the response time against T. gondii. Supernatants from macrophage cultures or T. lewisi by rat did not induced any immunosuppression. Rev. Biol. Trop. 57 (1-2): 13-22. Epub 2009 June 30.

El efecto inmunosupresor de la infección de T. lewisi sobre la multiplicación de T. gondii fue comparado en macrófagos peritoneales (MP) y alveolares (MA) de rata. El número de parásitos (taquizoitos) intracelulares (TI) fue contado por microscopía de luz. Los macrófagos alveolares y peritoneales (MP) de animales con 4 días de infección con T. lewisi muestran un aumento significativo en la multiplicación de T. gondii. A diferencia de los MP (que muestran un aumento en la multiplicación de T. gondii al cuarto día de infección con T. lewisi disminuyendo hacia el séptimo día), los MA mantienen un aumento en la multiplicación del parásito desde el cuarto, aumentando hacia el séptimo día de infección. Esta diferencia se puede deber a la ruta de infección utilizada para los experimentos que afectan directamente los MP donde se observa un efecto mayor y más temprano en comparación con los MA aislados de los pulmones, compartimiento afectado cuando la infección se vuelve sistémica entre el cuarto y sexto día de infección. La inmunidad contra T. gondii es similar entre ambas células fagocíticas, pero el tiempo de infección y el compartimiento donde se encuentren las células hace la diferencia en el tiempo de respuesta contra un parásito dado, en nuestro caso T. gondii. No hubo evidencia de que los sobrenadantes de cultivos de macrófagos provenientes de ratas infectadas ni el lisado de tripanosomas indujeran el efecto inmunosupresor.

Expression of Endothelin-1 by Stimulation with CXCL8 in Mouse Peritoneal Macrophages

Endothelin-1 (ET-1) has been characterized as a potent vasoconstrictor secreted by the endothelium, and play a major role in the regulation of vascular tone. It has been also known to participate in inflammatory reactions. The production of ET-1 by macrophages during infection and inflammation is related to tissue perfusion and leukocyte extravasation. The aim of this study is to investigate the role of IL-8/CXCL8, as a major inflammatory chemokine, for ET-1 expression in macrophges. Expression of ET-1 mRNA in mouse peritoneal macrophages (PeM phi) was weaker than that in vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). However, expression of IL-8/CXCL8-induced ET-1 mRNA in PeM phi was much more stronger than that in SHR and WKY VSMCs. Maximum expression of ET-1 mRNA was observed at 50 ng/ml dose of IL-8/CXCL8 and occurred at 2 h after addition of IL-8/CXCL8. Expression of ET-1 by IL-8/CXCL8 was dependent on NF-kappaB activation and ERK1/2 phosphorylation. Baicalein, a 12-lipoxygenase (LO) inhibitor, inhibited the expression of IL-8/CXCL8-induced ET-1 mRNA. This inhibitory action of baicalein was mediated via ERK1/2 inactivation. Induction of 12-LO mRNA by IL-8/CXCL8 and expression of ET-1 mRNA by 12-LO metabolite, 12(S)-HETE were also detected. The expression of IL-8/CXCL8-induced ET-1 mRNA was not detected in PeM phi transfected with 12-LO siRNA. These results suggest that IL-8/CXCL8 can act as one of main inducers of ET-1 in vascular inflammatory reactions, and ET-1 expression by IL-8/CXCL8 is related to 12-LO pathway in PeM phi.

Effect of Lipoteichoic Acid on Phagocytosis of Peritoneal Macrophages and NO Production in vitro / 华中科技大学学报(医学版)

Objective To explore the effects of lipoteichoic acid(LTA)on phagocytosis of peritoneal maerophages and NO production in vitro.Methods The peritoneal macrophages that were isolated sterilely from Kunming mice were divided into two groups:co-culture with RPMI 1640,and pretreatment with dexamethasone(DEX).Then,the LTA isolated from BFA with different final concentrations(5,10,20 and 40 ug/ml) was added.After the peritoneal macrophages were cultured for 4 and 24 h,the effects of LTA on cytophagocytesis and NO production in vitro were determined respectively by using neutral red and NO assay kit.Results LTA with final concentrations of 5,10,20 and 40/μg/mL significantly promoted the phagocytosis of peritoneal macrophages and production of NO(P<0.05).It also effectively antagonized the inhibitory response of DEX against peritoneal macrophages(P<0.05).The higher concentration of LTA,the stronger effect.Conclusion Not only can LTA stimulate and activate the peritoneal macrophages significantly,but also antagonize the inhibition from DEX in vitro.

Ca(OH)2 action on TNF-alpha and NO release in macrophage culture stimulated by Pseudomonas aeruginosa LPS / Ação do Ca(OH)2 sobre a produção de TNF-alfa e NO de cultura de macrófagos estimulada por LPS de Pseudomonas aeruginosa

Purpose: Calcium hydroxide [Ca(OH)2] interaction with the immune system to destroy or neutralize bacteria and their by-products is not completely understood. This study evaluated the calcium hydroxide ability to neutralize Pseudomonas aeruginosa lipopolysaccharides (LPS) using two different methods: nitric oxide (NO) stimulation and Tumoral Necrosis Factor-alpha (TNF-alpha) release in mice macrophage culture. Methods: For the NO assay, peritoneal exudate cells (PECs) were placed in contact with 25ìg/mL and 50ìg/mL LPS and LPS/Ca(OH)2 suspensions (50ìg/25mg and 25ìg/25mg). After incubation for 8h, Griess reagent was used, and the NO release was quantified. In the TNF-alpha assay the LPS solution was tested in 25ìg/mL, and the LPS/Ca(OH)2 suspension was tested in 25ìg/25mg concentration. After incubation for 24 hours, cells were fixed and stained with crystal violet. Absorbance values were obtained. Results were expressed in micromols. All tests were performed in triplicate. Results: The presence of Ca(OH)2 in both tested concentrations significantly reduced NO and TNF-alpha release. Conclusion: It can be concluded that bacteria LPS is a strong stimulus for NO and TNF-alpha release, but calcium hydroxide can neutralize it.

Objetivo: A ação do hidróxido de cálcio [Ca(OH)2] com o sistema imune e o mecanismo de neutralização das bactérias e seus subprodutos ainda não foi completamente esclarecida. Neste estudo foi avaliada a capacidade do Ca(OH)2 em neutralizar o lipopolissacarídeo (LPS) de Pseudomonas aeruginosa, utilizando-se duas metodologias: liberação de Óxido Nítrico (NO) e Fator de Necrose Tumoral Alfa (TNF-alfa) em cultura de macrófagos peritoneais de camundongos. Metodologia: No ensaio do NO, as células peritoneais foram expostas a uma solução de LPS (25mg/mL e 50mg/mL); e à suspensão de LPS/Ca(OH)2 em duas concentrações (50mg/ 25mg e 25mg/25mg). Após 8 horas de incubação, foi utilizado reagente de Griess, e a liberação de NO foi quantificada. No ensaio do TNF-alfa, a solução de LPS foi usada na concentração de 25mg/mL e o LPS/Ca(OH)2 a 25mg/25mg. Após 24 horas, as células foram fixadas e coradas com cristal violeta, e os valores de absorbância foram obtidos. Os resultados foram expressos em micromols. Todos os testes foram realizados em triplicata. Resultados: A presença de Ca(OH)2 nas duas concentrações avaliadas reduziu significativamente a liberação de NO e TNF-alfa. Conclusão: Pode-se concluir que o LPS bacteriano representa um forte estímulo para liberação destas citocinas, mas o hidróxido de cálcio foi capaz de neutralizar este efeito.

Inhibition of pathogenic Leptospira on phagocytosis of peritoneal macrophages of guinea pigs / 上海第二医科大学学报

Objective To investigate the effects of different Leptospira strains on phagocytosis of peritoneal macrophages of guinea pigs,and explore the role of innate immune in the pathogenesis of leptospirosis. Methods Peritoneal macrophages of guinea pigs were infected in vitro by three different Leptospira strains,the virulent Leptospira interrogans serovar Lai type strain Lai,the avirulent L.interrogans serovar Lai type strain IPAV,and the nonpathogenic L.biflexa serovar Patoc type strain PatocⅠ,respectively,and heat inactivated Staphylococcus epidermidis was added 0.5,1.5,3 and 6 h after infection and incubated for 30 min.The effect of Leptospira on the phagocytosis of macrophage was evaluated by the inactivated Staphylococcus epidermidis phagocytosis rate and phagocytosis index.Phagocytosis and ultrastructure of peritoneal macrophages were observed by transmission electron microscopy 3 h after infection,and changes of cytoskeleton of the macrophages were observed by laser scanning confocal microscopy. Results The phagocytic rates and phagocytic indexes of strain Lai,strain IPAV and strain PatocⅠinfection groups were significantly lower than those of control group 3 h and 6 h after infection(P

Effect of ginkgolide B on the platelet-activating factor induced changes of chemotaxis and cytoskeleton of macrophages / 药学学报

Aim To study the inhibitory effect of ginkgolide B (BN52021) on the PAF induced changes of chemotaxis of murine peritoneal macrophages and the related polymerization of F-actin.Methods Chemotaxis assays were performed using a modified 48-well Boyden chamber. Actin polymerization of murine peritoneal macrophages was analyzed by flow cytometry using a specific fluorescent stain. Results Peritoneal macrophages significantly migrated toward platelet-activating factor(PAF) through a micropore filter; however, in the presence of PAF receptor antagonist BN52021 (0. 01the actin polymerization of murine peritoneal macrophages induced by PAF in the presence of Ca2+ , but not in Ca2+ -free medium. Conclusion The results suggested that preventing polymerization of F-actin may be a pathway by BN52021 to inhibit the chemotaxis of macrophages, and this effect seems to be Ca2+dependent. The data further indicated that inhibition of PAF induced macrophage chemotaxis is an important mechanism underlying the anti-inflammatory action of BN52021.

The effects of ganoderma lucidum polysaccharides peptide (GLPP) on the nitric oxide production in mice peritoneal macrophages / 中国药理学通报

Aim To study the effects of Ganoderma polysaccharide peptide (GLPP) on the production of nitric oxide (NO) in mice peritoneal macrophages induced by LPS and its mechanism. Methods The effects of GLPP on the NO releasing from mice peritoneal macrophages induced by LPS were detected by using Griess reagent and the expression of iNOS by GLPP in mice peritoneal macrophages was detected by immunohistochemical method.Result The production of NO was increased by GLPP.(25～200 mg?kg -1) ig for five days or by GLPP(3.125～200 mg?L -1) in vitro but the effects of LPS on the production of NO was not influenced significantly. The expression of iNOS was increased by GLPP(25～200 mg?kg -1) ig for five days or GLPP(3.125～200 mg?L -1) in vitro.Conclusion GPP in vivo or in vitro could increase the production of NO in mice peritoneal macrophages. It might take effects by enhancing the synthesis of iNOS.

Inhibitory effects of sinomenine on the activation of nuclear factor-?B and on the degradation of I?B / 中国药理学通报

Aim To observe the effect of Sinomenine on the activation of NF-?B and the degradation of its inhibitor I-?B in peritoneal macrophages of BALB/c mice in vitro and provide experimental evidence for further evaluation of the antiinflammatory and antirheumatic effects of Sinomenine. Methods Effect of Sinomenine on the activation of NF-?B p65 in the cells was investigated by using fluorescence-labelling and laser confocal scanning microscopy; Effect of Sinomenine on the degradation of I?B-? was investigated by Western blot. Results SIN (0.25,1.25 mmol?L -1) attenuated the activation of NF-?B p65 in peritoneal macrophages of BALB/c mice induced by LPS in vitro. SIN(0.25,1.25 mmol?L -1) inhibited the degradation of I-?B-? in peritoneal macrophages of BALB/c mice induced by LPS in vitro. Conclusion SIN can partially inhibit the activation of NF-?B p65 and the degradation of I?B-? in peritoneal macrophages of BALB/c mice induced by LPS in vitro.

Study on the enhanced anti-tumoral immune function of human peritoneal macrophages by Sapylin / 中国免疫学杂志

Objective:To investigate the mechanism for intraperitoneal injection of sapylin to enhance the anti-tumoral immune functions of peritoneal macrophages.Methods:72 cases of patients with early or middle stage gastrointestinal cancer were peritoneally injected saline or sapylin, 48 h or 24 h before operation. The peritoneal macrophages(PM?) was harvested during their operations. The number of the PM? was counted. The phagocytosis, the enzyme activity of LDH and ACP,and the NO secretion were analyzed. Using human gastric cancer cell line MKN1 as target cells, the anti-tumoral cytotoxicity of the PM? was observed.The greater omentum was harvested,both the number and the size of the omental milky spots were measured.Results:I.p. injection of sapylin significantly increased the number, the phagocytosis, the enzyme activities,NO secretion and anti-tumoral cytotoxicity of the PM?, as well as the number and the size of omental milky spots.Conclusion:By activating omental milky spots, intraperitoneal injection of sapylin increased the number and enhanced the immune function of PM?, the activated PM? showed enhancement of anti-tumoral cytotoxicity.

Free radical scavenging activity by Ganoderma lucidum polysaccharides peptide on peritoneal macrophages in mice / 中国临床药理学与治疗学

AIM: To investigate the free radical scavenging activity of Ganoderma lucidum polysaccharides peptide (GLPP) on peritoneal macrophages in mice. METHODS: Alloxan and tert butylhydroperoxide (tBOOH) were used as an oxidant to injury peritoneal macrophages in vivo in mice or in vitro, respectively. 2', 7' dichlorodihydrofluorescein diacetate (DCHF DA) was used as fluorescent probe. The fluorescence from cells was observed under the laser confocal microscope. Time series scan of conforcal microscope was used to observe the changes of fluorescence by GLPP in mice peritoneal macrophages over time. RESULTS: The results of confocal microscopy showed that GLPP (100 mg?kg -1 , ig for 5 d ) lowed fluorescence in the mice macrophages injured by alloxan (75 mg?kg -1 iv). GLPP (10 mg?L -1 ) also lowed fluorescence in the mice macrophages injured by tBOOH ( 7.76 ?10 -5 mol?L -1 ) in vitro as well. Time series scan showed that GLPP (10 mg?L -1 ) lowed fluorescence in the mice macrophages at rest state or during the respiratory burst induced by PMA (50 nmol?L -1 ). CONCLUSION: GLPP shows antioxidant effects and might have free radical scavenging effects on peritoneal macrophages in mice.