Macrophages In Peritoneum Research Articles

Immunomodulatory Activity of Yogurt Fortified with Honey and Rosella (Hibiscus sabdariffa L.) on Reactive Oxygen Intermediate (ROI) and Nitric Oxide (NO) Secretion

Raise the body's immune system. Rosella (Hibiscus sabdariffa Linn.) is known to have anthocyanin compounds that have antioxidant and immunomodulatory effects. The aim of this research is to know the effect of immunomodulator yogurt which fortified by rosella ( Hibiscus sabdariffa L.) extract to increase the secretion of Reactive Oxygen Intermediate (ROI) and Nitric Oxide (NO). This research was conducted in vivo using 25 male mices with Balb/C strain divided into 5 groups: normal group, plain yogurt treated group and 2%, 4%, 8% rosella yogurt treated group. Treatment was given for 21 days orally. On the 22nd day the mice were sacrificed by taking peritoneum macrophage cells and then tested the secretion of Reactive Oxygen Intermediate (ROI) and Nitrite Oxide (NO). The results showed that there was an increase of ROI and NO secretion in 2%, 4%, and 8% rosella yoghurt treated groups compare to plain yoghurt. The fortification of yoghurt with rosella extract and honey increase the potency of yoghurt in increasing the immunomodulatory activity

Aspirin inhibits LPS-induced macrophage activation via the NF-\u03baB pathway

Aspirin (acetylsalicylic acid, ASA) has been shown to improve bone marrow mesenchymal stem cell-based calvarial bone regeneration by promoting osteogenesis and inhibiting osteoclastogenesis. However, it remains unknown whether aspirin influences other immune cells during bone formation. In the present study, we investigated whether ASA treatment influenced macrophage activation during the LPS inducement. We found that ASA could downregulate the expressions of iNOS and TNF-α both in mouse peritoneum macrophages and RAW264.7 cells induced by LPS via the IκK/IκB/NF-κB pathway and a COX2/PGE2/EP2/NF-κB feedback loop, without affecting the expressions of FIZZ/YM-1/ARG1 induced by IL-4. Furthermore, we created a rat mandibular bone defect model and showed that ASA treatment improved bone regeneration by inhibiting LPS-induced macrophage activation in the early stages of inflammation. Taken together, our results indicated that ASA treatment was a feasible strategy for improving bone regeneration, particularly in inflammatory conditions.

Effect of HylocereusPolyrhizus Rind Extract Toward Interleukin-1β, Vascular Endothelial Growth Factor Expression, Endometriosis Implant Area

Retrograde menstruation lead to I Kappa B Kinase (IKK) fosforilation in peritoneum macrophage and cause secretion of proinflammatory cytokine interleukin1β then stimulate endometriosis cell to produce Vascular Endothelial Growth Factor which lead to increasing of endometriosis lession seen as endometriosis implant area. Cytokine secretion was inhibited through prevention of NF-κB activation by dragon red fruit rind extract (Hylocereuspolyrhizus). The aim of this reserach is to know the effect of dragon red fuit rind extract with 0,25; 0,5; and 1 mg/g bodyweight dosage toward IL-1β, VEGF expression and implant area in endometriosis mice model. The design of this experiment was randomized post test only control group design.Endometrios mice model were made in 14 days and split into two group, positive control group and treatment group after two week negative control group and postive control group were given Na-CMC 0,5% solution consequetively, and treatment group were given dragon red fruit extract with different dosage. Signification number for IL-1β is p>0,05, signification number for VEGF is p>0,05, and implant area signification number is p>0,05. Administration of dragon red fruit rind extract can decrease IL-1β, VEGF, and implant area.

Protectin DX increases survival in a mouse model of sepsis by ameliorating inflammation and modulating macrophage phenotype

Recently, a serial of studies have demonstrated that lipid mediators derived from Omega-3 fatty acid docosahexaenoic acid have pro-resolving or anti-inflammatory effects in many inflammatory diseases. Here, we sought to evaluate whether Protectin DX (PDX, an isomer of Protecin D1), a newly identified lipid mediator, could protect mice against sepsis and explore the underling mechanism. Animal model of sepsis was established by cecum ligation and puncture (CLP). We found that PDX increased overall survival rate within eight days and attenuated multiple organ injury in septic mice. In addition, PDX reduced pro-inflammatory cytokines and bacterial load 24 h after CLP. Moreover, PDX promoted phagocytosis of peritoneal macrophages and increased the percentage of M2 macrophages in peritoneum of septic mice. In vitro, M2 macrophage markers (Arg1 and Ym1) and its transcriptional regulator (peroxisome proliferator-activated receptor-γ, PPAR-γ) were upregulated in Raw264.7 macrophages challenged with PDX. GW9662 (a PPAR-γ inhibitor) and PPAR-γ siRNA abrogated the induction of Arg1 and Ym1 by PDX in Raw264.7 cells. Taken together, our results suggest that PDX is able to promote M2 polarization, enhance phagocytosis activity of macrophage and accelerate resolution of inflammation, finally leading to increased survival rate of septic mice.

Abstract 588: Optimization of Radiolabeled Liposomes for in vivo Atherosclerosis Imaging by SPECT

Objectives: Molecular imaging enables non-invasive in vivo detection of vulnerable plaques. Last year, we reported an atherosclerosis imaging agent, 111 In-labeled liposome. The liposome was modified with phosphatidylserine (PS), since macrophages recognize PS to phagocytize apoptotic cells in the plaque. PS modification was successful and we could visualize atherosclerotic plaques by SPECT. However, the too quick blood clearance seemed to have reduced PS-liposome accumulation to the plaque. Therefore, in this study, we PEGylated the liposome to retard the blood clearance. Methods: PS liposomes were prepared by lipid-film hydration method. The liposomes, sizes in 100 or 200 nm, were PEGylated with PEG2000 or PEG5000 in 1 or 5% w/w. The liposomes were radiolabeled by encapsulating 111 In-NTA using active-loading method. For in vitro uptake study, the liposomes were incubated with mouse peritoneum macrophages. The in vivo biodistribution studies were done in ddY mice. En face autoradiograms were obtained with ApoE-/- mice 2 hr after the liposome injection. For SPECT imaging, WHHL rabbits were used. Results: The in vitro uptake levels to macrophages were decreased by PEGylation. 5% PEG5000 decreased the uptake to the half of non-PEGylated one, but 1% PEG2000 affected less than 5%. In vivo results showed slower blood clearance in PEGylated liposomes (12 %dose/g for 5%PEG5000, 2.0 %dose/g for 1%PEG2000, and 1.0 %dose/g for non-PEGylated liposome at 60 min after the injection). PS modification improved target-to-nontarget ratios, and 5%PEG2000 added 200 nm liposome showed the highest uptake to the region. The aorta was successfully visualized by SPECT at 48hr after the 111 In-labeled 5%PEG2000-PS200 liposome injection. Conclusions: PEGlylation prolonged the blood clearance, but decreased the uptake to macrophages. However, the effect was larger for blood clearance and decrease in macrophage uptake was acceptable for in vivo imaging.

Abstract 145: Development of Radiolabeled Liposomes for Atherosclerotic Plaque Imaging

Objectives Molecular imaging enables non-invasive in vivo detection of vulnerable plaques. Macrophage infiltration is characteristics for atherosclerotic vulnerable plaques, and many apoptotic cells are seen in this region. Also, it is known that macrophages recognize phosphatidylserine (PS) that is exposed on the cell surface during apoptosis to phagocytize these cells. In this study, we prepared radiolabeled phosphatidylserine liposomes for detection of vulnerable plaques. Methods PS liposomes and control liposomes (phosphatidylcholine (PC) liposomes) were prepared by lipid film hydration. The prepared sizes were 100 nm and 200 nm (PS100, PS200, PC100, and PC200 liposomes). The liposomes were radiolabeled by encapsulating 111 In-NTA using active-loading method. The 111 In-liposomes were incubated with mouse peritoneum macrophages for 2hr and uptake level was investigated. For biodistribution study, 111 In-liposomes were intravenously injected to ddY mice. Also, the 111 In-liposomes were injected to apoE -/- mice, and the aortas were harvested for autoradiography and Oil-Red O staining. For SPECT imaging, WHHL rabbits were used. Results The in vitro uptake levels to macrophages were 60.5, 14.7, 32.0, 14.4 %dose/mg protein, for 111 In-PS100, 111 In-PC100, 111 In-PS200, 111 In-PC200, respectively. The liver uptake was high in all liposomes, and blood clearance was faster in PS liposomes than PC liposomes. The in vivo distributions of 111 In-labeled PS liposomes to atherosclerotic regions were well matched with Oil-Red O staining in apoE -/- mouse. Target-to-non-target ratio was highest in 111 In-PS200. The aorta was successfully visualized by SPECT at 48hr after the 111 In-labeled PS liposome injection. Conclusions By in vitro uptake study, it is revealed that macrophage targeting was accomplished by PS. Also, atherosclerotic region was successfully detected by 111 In-PS200 in apoE-/- mice and WHHL rabbits in vivo .

Phytochemical analysis, antioxidant and anti-inflammatory activities of Phyllanthus simplex

Ethnopharmacological relevancePhyllanthus simplex (Family: Euphorbiacae) is widely used in traditional medicines for treatment of various diseases including inflammation. Materials and methodsPetroleum ether extract (PSPE) and ethanol extract (PSEE) of the whole plant of Phyllanthus simplex were characterized for their total phenolics, tannins and flavonoids content. These extracts were standardized by HPTLC using phyllanthin and gallic acid respectively as markers. Antioxidant activity of extracts was evaluated by the DPPH, hydroxyl and superoxide radicals scavenging assay. The total antioxidant capacity of extracts was determined. Anti-inflammatory activity was evaluated by their effect on nitric oxide (NO) production in isolated rat peritoneal macrophages; carragennan-induced paw edema and formation of cotton pellet-induced granuloma in rats. ResultsAbundance of phenolics was found in PSEE. Phyllanthin and gallic acid content in PSPE and PSEE were found to be 14.5 and 0.65% (w/w) respectively. PSEE showed concentration dependent significant scavenging of DPPH, hydroxyl and superoxide radicals with IC50 values 102.219, 171.485 and 24.73μg/ml respectively. PSEE significantly inhibited NO production in isolated rat peritoneum macrophages. Moreover, it also exhibited significant inhibition of carragennan-induced paw edema (58.48±0.028%, p<0.001, at 6h, 200mg/kg oral dose) and cotton pellet-induced granuloma formation (45.671±0.712%, p<0.001, at 200mg/kg oral dose). Anti-inflammatory activity of PSEE was found to be comparable to diclofenac sodium. ConclusionsSignificant antioxidant and anti-inflammatory activities were found in PSEE which may be attributed to its high phenolic content.

Fatty acid composition, antioxidant, anti-inflammatory and antibacterial activities of seed oil from Crotalaria juncea Linn.

Chemical characteristics, fatty acid composition, antioxidant, anti-inflammatory and antibacterial activities of Crotalaria juncea seed oil (CJSPE) were evaluated in this study. High amount of linoleic acid (62.36%) was found in CJSPE by the gas liquid chromatography (GLC) study. Antioxidant activity of CJSPE was evaluated by in vitro assay methods which revealed the 2,2-Diphenyl-1-picrylhydrazyl (DPPH), hydroxyl and superoxide radical scavenging activity of CJSPE; its antioxidant activity was found to be concentration dependent and IC50 values were 132.31, 286.409 and 31.254 μg/ml respectively. Moreover, CJSPE has displayed dose dependant, significant inhibition of NO production in the isolated rat peritoneum macrophages; and significant anti-inflammatory activity in carragennan-induced paw edema (64.52 ± 0.053%, p < 0.001, at 6 h, 200 mg/kg oral dose) and cotton pellet-induced granuloma formation (48.55 ± 0.244%, p < 0.001, at 200 mg/kg oral dose) models of inflammation; and its anti-inflammatory effect was comparable to that of diclofenac sodium. However, moderate antibacterial activity of CJSPE was observed. In conclusion, the study demonstrated significant antioxidant and anti-inflammatory activities of CJSPE. Key words: Crotalaria juncea, antibacterial activity, anti-inflammatory, antioxidant activity, linoleic acid.

Peritoneal macrophage infiltration is correlated with baseline peritoneal solute transport rate in peritoneal dialysis patients

High baseline peritoneal solute transport rate is reportedly associated with reduced patient and technique survival in continuous peritoneal dialysis (PD) patients. However, the determinants of baseline peritoneal solute transport rate remain uncertain. The aim of this study was to investigate the relationship between peritoneal local inflammation, angiogenesis and systemic inflammation and baseline peritoneal permeability. Peritoneal biopsy specimens from 42 pre-dialysis uraemic patients and 11 control individuals were investigated. Immunohistochemistry for CD68-positive macrophages, chymase- and tryptase-positive mast cells, interleukin-6 (IL-6)-positive cells, CD3-positive T cells, CD20-positive B cells, neutrophils and CD31- and pathologische anatomie Leiden-endothelium (PAL-E)-positive blood vessels in the peritoneum was performed. Baseline dialysate-to-plasma ratio for creatinine (D/P Cr) was determined within 6 months of PD induction. Clinical and laboratory parameters were measured at the time of peritoneal biopsy. Factors associated with peritoneal permeability were assessed by multiple linear regression analysis. Pre-dialysis uraemic peritoneum showed infiltration by CD68-positive macrophages, and mast cells, as compared with controls. Baseline D/P Cr was correlated with density of CD68-positive macrophages (P < 0.001), IL-6-positive cells (P < 0.001), CD31-positive (P < 0.05) and PAL-E-positive blood vessels (P < 0.05) and serum albumin (P < 0.05). However, baseline peritoneal permeability was not correlated with infiltration by mast cells, B cells, T cells, neutrophils, serum C-reactive protein or other clinical factors. On multiple linear regression analysis, the number of CD68-positive macrophages in peritoneum was an independent predictor for baseline peritoneal permeability (P = 0.009). Peritoneal macrophage infiltration is predominant in uraemic patients and is an important factor in predicting baseline peritoneal permeability.

Immunomodulatory and In vivo Antiplasmodial Activities of Propolis Extracts

Problem statement: Malaria caused by various species of prozoan paras ites, Plasmodium . The exact mechanism of humoral immunity in eliminating Plasmodium is not completely understood. However, an antimalarial antibody is strongly suspe cted to play important role in immunity. The purpose of this research was to evaluate the immuno modulator and antiplasmodial activities of Indonesian propolis extracts. Approach: Thirty male BALB/c mice weighing approximately 25-30 g aged between 6 and 8 weeks old were used for pr opolis treatment in vivo . The mice infected with 0.1 mL of the P. berghei suspension at a concentration of 10 7 parasites per mouse on day 0. The drug was administered orally for 4 days from day 0-3 at doses 25, 50 and 100 mg kg -1 BW in the experimental group. The control group was given the solven in equal volume for the same duration. This research utilized not only hypersensitivity re action which measures the humoral immunity by SRBC-immunized mice but also the activity and capacity of peritoneum macrophage phagocytosis in Plasmodium berghei-infected mice. The parasitaemia number was calculated using blood smear method every day for 4 day after the mice had been infected P. berghei to identify the antiplasmodial activity. Results: The research results revealed that Propolis Hydroa lcoholic Solution (PHS) had a strong immunomodulatory activity but weak antiplasmodial activity. Conclusion: It was concluded that PHS showed more immunostimulant activity than antiplasmodial activity, proved by the increase of IgG and the macrophage phagocytosis activity and capacity. The antiplasmodial activity of PHS was due to the mice immunity increase so that they lived longer.

Efeito da Mannheimia granulomatis sobre cultivo de fibroblastos

Primary cultures of Mannheimia granulomatis were established in chicken embryos to assess their capacity to stimulate fibroblast proliferation. The capacity of the bacterium-activated macrophages to stimulate cytokine and enzyme proliferation was assessed in a mouse peritoneum macrophage culture. To evaluate the bacteria infection on fibroblasts and their growth within 48h in relation to the active macrophages, cultures were washed and trypsinized and the cells counted. Results showed no significant differences when the bacteria-infected fibroblasts were mixed with bacterial extract (P=0.9682). The treatment using just products of macrophages resulted similar to the negative control. Significant differences on cell proliferation were established (P=0,0039) when the products of M. granulomatis-activated macrophages were used, meaning that bacterial components were unable to promote fibroblast increase. Further research is needed to elucidate the effect of M. granulomatis on the macrophages.

VAKSINASI BCG MENINGKATKAN AKTIVITAS FAGOSITOSIS DAN SEKRESI REACTIVE OXYGEN INTERMEDIATE (ROI) PADA MAKROFAG PERITONEUM KUCING YANG DIINFEKSI DENGAN Mycobacterium tuberculosis

ABSTRACT The aim of this experiment is to study the effect of BCG vaccination on phagocyte activity and ROI secretion in cat peritoneum macrophages which infected with M.tuberculosis. The experiment used twenty four healthy cats. The animals were divided in 2 groups, 12 cats in each group. Group I were vaccinated with BCG, group II were control group which unvaccinated. BCG vaccination was done twice in two weeks interval. Two days after vaccination each cat was infected by 105 cfu M.tuberculosis intraperitoneally. The activity of macrophages were measured  at 1st, 2nd, 12th, and 24th, after infection using in vitro latex bead phagocyte and NBT reduction assay. Three cats were used to measure the macrophage activity in each period, using triplicate sample for each cat. The results of the experiment showed that the phagocyte activity and ROI secretion increased significantly in vaccination group (P&lt;0.01) compared with the control group, and these activities reached to the plateau level at 2 weeks after infection. Although these enhanced activities were gradually diminished thereafter, higher levels of these activities were consistently observed until the end of experiment compared with control group. BCG vaccination increased the cellular immunity especially phagocyte and ROI secretion activities of peritoneual macrophages in cat infected with M.tuberculosis Key words: BCG, phagocyte, ROI, M.tuberculosis, cat

VAKSINASI BCG MENINGKATKAN AKTIVITAS FAGOSITOSIS DAN SEKRESI REACTIVE OXYGEN INTERMEDIATE (ROI) PADA MAKROFAG PERITONEUM KUCING YANG DIINFEKSI DENGAN Mycobacterium tuberculosis

ABSTRACT The aim of this experiment was to study the effect of BCG vaccination on phagocyte activity and ROI secretion in cat peritoneum macrophages which infected with M.tuberculosis. The experiment used twenty four healthy cats. The animals were divided in 2 groups, 12 cats in each group. Group I were vaccinated with BCG, group II were control group which unvaccinated. BCG vaccination was done twice in two weeks interval. Two days after vaccination each cat was infected by 105 cfu M.tuberculosis intraperitoneally. The activity of macrophages were measured  at 1st, 2nd, 12th, and 24th, after infection using in vitro latex bead phagocyte and NBT reduction assay. Three cats were used to measure the macrophage activity in each period, using triplicate sample for each cat. The results of the experiment showed that the phagocyte activity and ROI secretion increased significantly in vaccination group (P&lt;0.01) compared with the control group, and these activities reached to the plateau level at 2 weeks after infection. Although these enhanced activities were gradually diminished thereafter, higher levels of these activities were consistently observed until the end of experiment compared with control group. BCG vaccination increased the cellular immunity especially phagocyte and ROI secretion activities of peritoneual macrophages in cat infected with M.tuberculosis Key words: BCG, phagocyte, ROI, M.tuberculosis, cat

In Vitro Studies of Cytotoxic Effect on Sarcoma-180 Cells of .BETA.-Subunit from Egg White Ovomucin.

When sarcoma-180 (SR-180) cells were incubated with α- or β-subunit prepared from insoluble ovomucin in the gel fraction of egg white, the cell proliferation rate was apparently reduced by the addition of β-subunit, while it was largely unaffected by α-subunit. Examinations by scanning and transmission electronmicroscopy indicated a dose- and time-dependent cytotoxic effect of β-subunit on SR-180 cells, whereas β-subunit did not injure mouse peritoneum macrophage used as normal cells. Ultrastructurally, SR-180 cells treated with β-subunit showed changes such as swelling and bleb formation of microvilli on cell membrane, irregular clumping of chromatin, irregular nuclear shape, and marked swelling of organelles in cytoplasm associated with cell degeneration in necrotic change. Furthermore, apoptotic analysis of the cultured SR-180 cells also demonstrated that β-subunit did not induce the apoptotic DNA fragmentation.

In vivo indomethacin reverse exercise-induced immunosuppression in rats

The effect of oral indomethacin on the immunosuppressive effect of exercise was examined in exercised untrained female Wistar rats immunized with sheep red blood cell (SRBC) antigens. Intensity of the 1 h exercise was controlled by 0–50 kPa air pressure, generated by a compressor located at the bottom of a water tank, during continuous swimming of the rats, previously immunized with SRBC. After 48–72h, depending on the ip (intraperitoneal) or iv (intravenous) route of SRBC immunization, the exercise suppressed humoral PFC response and augmented phagocytosis of peritoneum macrophages. These effects occurred only when exercise was performed at 48 h after antigen injection. Animals receiving indomethacin, however, did not show any exercise-related suppression of the PFC response. The data suggest a relationship between exercise-induced immunosuppression and possible increased in vivo prostaglandin synthesis during the intense exercise. Overall, exercise-related suppression of humoral PFC response was dependent on the intensity of the exercise, was time specific, and was reversible by pharmacological blockade of the cyclooxygenase pathway of prostaglandin synthesis.