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1.
Acta cir. bras ; 31(2): 119-125, Feb. 2016. graf
Article in English | LILACS | ID: lil-775562

ABSTRACT

PURPOSE: To detect whether chitin and sepia ink sponge (CS) can promote wound healing and elevate impact of CS on phagocytosis ability of macrophages. METHODS: Forty-eight rats were assigned to four groups: Normal group (Normal), negative control group (Con), chitin and sepia ink sponge group (CS) and positive control Surgicel Gauze(r) group (SG). Deep second-degree burn model was created in rats. Wound area was recorded by digital imaging and determined using Image J software. Samples were collected and kept at -80oC on 3d, 7d, 14d and 21d for cytokines detecting. Transforming growth factor (TGF)-β1, interleukin (IL)-6, matrix metalloproteinase (MMP)-1, hydroxyproline (Hyp) and macrophage activity reflected by tumor necrosis factor (TNF)-α were determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Comparing to Con and SG, scabs in CS group fell off and basically healed on 21 day. TGF-β1, IL-6, MMP-1 and Hyp were significantly increased by CS and SG comparing to Con (p < 0.05), CS had more apparently adjustment on TGF-β1 and MMP-1 compared to SG; results in vitro indicated CS significantly promoted phagocytosis ability of macrophages reflected in TNF-α (p < 0.05). CONCLUSION: CS improved wound healing through exerting significant influences on secretion of kinds of cytokines and activating macrophages.


Subject(s)
Animals , Male , Wound Healing/drug effects , Burns, Chemical/drug therapy , Chitin/pharmacology , Sepia , Macrophages/drug effects , Phagocytosis/drug effects , Random Allocation , Chitin/therapeutic use , Cytokines/drug effects , Cytokines/metabolism , Rats, Wistar , Matrix Metalloproteinase 1/drug effects , Disease Models, Animal , Hydroxyproline/metabolism , Ink , Macrophages/metabolism
2.
Yonsei Medical Journal ; : 283-290, 2016.
Article in English | WPRIM | ID: wpr-147364

ABSTRACT

Macrophages (Mphi) play a pivotal role in the protection system by recognizing and eliminating invading pathogenic bacteria. Phagocytosis and the killing of invading bacteria are major effector functions of Mphi. Although the phagocytic and bactericidal activities of Mphi have been analyzed via several methods using a light microscope, a fluorescence microscope, or a fluorescence-activated cell sorter, expensive materials and equipment are usually required, and the methods are rather complicated. Moreover, it is impossible to determine both the phagocytic and bactericidal activities of Mphi simultaneously using these methods. In this review, we describe a simple, reproducible, inexpensive, yet old-fashioned method (antibiotic protection assay) for determining the phagocytic and bactericidal activities of Mphi.


Subject(s)
Anti-Bacterial Agents/pharmacology , Gentamicins/pharmacology , Macrophages/drug effects , Phagocytosis/drug effects
3.
Indian J Exp Biol ; 2015 Mar; 53(3): 158-163
Article in English | IMSEAR | ID: sea-158406

ABSTRACT

Chyawanprash is an ayurvedic formulation used in Indian traditional medicinal system for its beneficial effect on human health. We investigated the immunostimulatory effects of Chyawanprash (CHY) using in vitro assays evaluating the secretion of cytokines such as Tumor Necrosis Factor-alpha (TNF-α), Interleukin-1beta (IL-1β) and Macrophage Inflammatory Protein-1-alpha (MIP-1-α) from murine bone marrow derived Dendritic Cells (DC) which play pivotal role in immunostimulation. The effects of CHY on phagocytosis in murine macrophages (RAW264.7) and Natural Killer (NK) cell activity were also investigated. At non-cytotoxic concentrations (20–500 µg/ml), CHY enhanced the secretion of all the three cytokines from DC. CHY also stimulated both, macrophage (RAW264.7) as well as NK cell activity, in vitro. In conclusion, the data substantiates the immunoprotective role of CHY at cellular level mediated by immunostimulation in key immune cells viz. dendritic Cells, macrophages and NK cells.


Subject(s)
Adjuvants, Immunologic/pharmacology , Animals , Cell Line , Cytokines/analysis , Cytotoxicity, Immunologic/drug effects , Dendritic Cells/drug effects , Drug Evaluation, Preclinical , In Vitro Techniques , Killer Cells, Natural/drug effects , Macrophages/drug effects , Male , Medicine, Ayurvedic , Mice , Mice, Inbred C57BL , Phagocytosis/drug effects , Plant Preparations/pharmacology , Specific Pathogen-Free Organisms , Spleen/cytology , Zymosan
4.
Mem. Inst. Oswaldo Cruz ; 110(1): 56-64, 03/02/2015. tab, graf
Article in English | LILACS | ID: lil-741619

ABSTRACT

Histology is the gold standard for diagnosing acute rejection and hepatitis C recurrence after liver transplantation. However, differential diagnosis between the two can be difficult. We evaluated the role of C4d staining and quantification of hepatitis C virus (HCV) RNA levels in liver tissue. This was a retrospective study of 98 liver biopsy samples divided into four groups by histological diagnosis: acute rejection in patients undergoing liver transplant for hepatitis C (RejHCV+), HCV recurrence in patients undergoing liver transplant for hepatitis C (HCVTx+), acute rejection in patients undergoing liver transplant for reasons other than hepatitis C and chronic hepatitis C not transplanted (HCVTx-). All samples were submitted for immunohistochemical staining for C4d and HCV RNA quantification. Immunoexpression of C4d was observed in the portal vessels and was highest in the HCVTx- group. There was no difference in C4d expression between the RejHCV+ and HCVTx+ groups. However, tissue HCV RNA levels were higher in the HCVTx+ group samples than in the RejHCV+ group samples. Additionally, there was a significant correlation between tissue and serum levels of HCV RNA. The quantification of HCV RNA in liver tissue might prove to be an efficient diagnostic test for the recurrence of HCV infection.


Subject(s)
Animals , Humans , Mice , Annexin A1/pharmacology , Macrophages/drug effects , Macrophages/immunology , Neutrophils/cytology , Neutrophils/immunology , Apoptosis , Actins/metabolism , Annexin A1/deficiency , Annexin A1/genetics , Annexin A1/immunology , Cyclic AMP-Dependent Protein Kinases/metabolism , Cyclic AMP/metabolism , Dexamethasone/pharmacology , In Vitro Techniques , /biosynthesis , Mice, Knockout , Macrophages/metabolism , Peptides , Phagocytosis/drug effects , Transforming Growth Factor beta/biosynthesis
5.
Indian J Exp Biol ; 2014 Aug; 52(8): 808-813
Article in English | IMSEAR | ID: sea-153763

ABSTRACT

Guduchi has been widely used in the traditional medicine as an immunomodulator. Description of guduchi in Ayurvedic literature resemble with T. sinensis rather than with commonly available T. cordifolia and hence this may be used as substitutes for T. sinensis. T. cordifolia growing on Azadirachta indica commonly called Neem-guduchi has more immunomodulatory potential. Thus, immunomodulatory activity of three Tinospora spp. was assessed by checking humoral and cell mediated immune responses to the antigenic challenges with sheep RBCs and by neutrophil adhesion tests on albino Wistar rats using Guduchi-Satwa, a well known dosage form. Results revealed that Neem-guduchi possesses higher immunomodulatory potential at the dose of 300 mg/kg, po and validated the traditional claim. Hence, Neem-Guduchi can be employed in immunomodulatory formulation prepared using guduchi.


Subject(s)
Animals , Azadirachta/chemistry , Azadirachta/growth & development , Immunomodulation , Neutrophils/drug effects , Neutrophils/immunology , Phagocytosis/drug effects , Phagocytosis/immunology , Plant Extracts/administration & dosage , Plant Extracts/chemistry , Plant Extracts/immunology , Rats , Tinospora/chemistry , Tinospora/immunology
6.
Rev. méd. Chile ; 141(9): 1150-1157, set. 2013. ilus, graf, tab
Article in Spanish | LILACS | ID: lil-699682

ABSTRACT

Background: The rainforest is an important source of natural compounds with therapeutic properties. Although there are many anti-inflammatory and antineoplastic drugs available to the clinician, there is an ongoing need for new therapeutic drugs with fewer serious adverse effects. Aim: To evaluate the in vitro cytotoxic effects of lupeol and casearin G on tumor cells, on phagocytic activity and nitric oxide (NO) production by blood mononuclear cells. Material and Methods: The cytotoxic effect of these compounds on cell lines MCF-7 (human breast adenocarcinoma) and PC-3 (human prostate cancer) was measured by a colorimetric assay (MTS/PMS) and the sulphorhodamine B assay. Peripheral blood mononuclear cells were obtained from eight healthy volunteers. The effect of these compounds on nitric oxide (NO) production was measured using the Griess reaction. Their effect on phagocytic activity of PBMC was also evaluated. Results: Lupeol (≥ 2 mM) resulted in a reduction of both the phagocytic index and the percentage of phagocytic monocytes and macrophages. Treatment of monocytes/macrophages with lupeol (72 µM) and casearin G (4 µM) reduced the production of NO. Neither lupeol (< 969 µM) nor casearin G (< 55 µM) had cytotoxic effects on PBMC. Casearin G showed both cytotoxic (IC50, LC50) and cytostatic (GI50) effects against tumor cells, PC-3 (IC50 = 12.5 µM; GI50 = 13.3 µM; LC50 = 51.9 µM) and MCF-7 (IC50 = 112.8 µM; GI50 = 11.8 µM; LC50 = 49.4 µM), as well as a hemolytic effect (≥ 182 µM). Conclusions: These observations indicate that lupeol and casearin G might be useful compounds in the preparation of anti-inflammatory drugs, whereas casearin G might be useful in the elaboration of antitumor drugs.


Subject(s)
Humans , Anti-Inflammatory Agents/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Diterpenes/pharmacology , Leukocytes, Mononuclear/drug effects , Nitric Oxide/biosynthesis , Pentacyclic Triterpenes/pharmacology , Phagocytosis/drug effects , Antineoplastic Agents, Phytogenic/isolation & purification , Casearia/chemistry , Cell Line, Tumor/drug effects , Drug Screening Assays, Antitumor , Diterpenes/isolation & purification , Pentacyclic Triterpenes/isolation & purification , Zanthoxylum/chemistry
7.
Article in English | WPRIM | ID: wpr-65169

ABSTRACT

The incidence of diabetes mellitus is increasing among companion animals. This disease has similar characteristics in both humans and animals. Diabetes is frequently identified as an independent risk factor for infections associated with increased mortality. In the present study, homozygous diabetic (db/db) mice were infected with Listeria (L.) monocytogenes and then treated with the anti-diabetic drug exendin-4, a glucagon-like peptide 1 analogue. In aged db/db mice, decreased CD11b+ macrophage populations with higher lipid content and lower phagocytic activity were observed. Exendin-4 lowered high lipid levels and enhanced phagocytosis in macrophages from db/db mice infected with L. monocytogenes. Exendin-4 also ameliorated obesity and hyperglycemia, and improved ex vivo bacteria clearance by macrophages in the animals. Liver histology examined during L. monocytogenes infection indicated that abscess formation was much milder in exendin-4-treated db/db mice than in the control animals. Moreover, mechanistic studies demonstrated that expression of ATP binding cassette transporter 1, a sterol transporter, was higher in macrophages isolated from the exendin-4-treated db/db mice. Overall, our results suggest that exendin-4 decreases the risk of infection in diabetic animals by modifying the interaction between intracellular lipids and phagocytic macrophages.


Subject(s)
ATP-Binding Cassette Transporters/metabolism , Age Factors , Animals , Blood Chemical Analysis , Cholesterol/metabolism , Diabetes Mellitus, Type 2/drug therapy , Dyslipidemias/drug therapy , Female , Hyperglycemia/drug therapy , Hypoglycemic Agents/therapeutic use , Injections, Intraperitoneal , Lipid Metabolism/drug effects , Listeria monocytogenes/drug effects , Listeriosis/drug therapy , Macrophages/drug effects , Mice , Obesity/drug therapy , Peptides/therapeutic use , Phagocytosis/drug effects , Venoms/therapeutic use
8.
Egyptian Journal of Hospital Medicine [The]. 2011; 45 (October): 477-503
in English | IMEMR | ID: emr-145538

ABSTRACT

Tramadol is a synthetic opioid analgesic. It is commonly prescribed for moderate to severe pain, becoming abused more popular among teens in most countries. Paracetamol as anti-inflammatory drugs [acetaminophen] [APAP] is widely used as an analgesic and antipyretic agent. Meanwhile, tramadol/acetaminophen [tramacet] is effective in acute or chronic moderate-to-moderately severe pain. In comparative study, the current investigation threw the light on the effect of over doses of tramadol and/or APAP on the immune function and hepatocytes in adult male Sprague-Dawley rats. Treated rats received oral doses of each drug for 15 consecutive days and after last treatment, they kept three days later for withdrawal studies. The rats were divided into four treatment groups, in the first group, rats received saline and used as control. The second, third and fourth groups treated with tramadol [45 mg/kg], tramadol/APAP [45/450 mg/kg], APAP [450 mg/kg] respectively, once a-day at the first week and ending with 90, 90/900, 900 mg/kg at the second week. Rats were sacrificed at the end of the first, second weeks and three days of last treatment. Daily doses of tramadol and /or APAP exposure in rats decreased the cellularity of spleen. Moreover, phagocytic and killing of S. aureus by PMN and macrophage cells caused a highly significant decrease in treated groups. IFN-gamma was reduced in a statistically different treated group of rats. Serum IL-10 was unaffected by any of the treatment regimens but increased only in tramadol/APAP treated rats. Spleen histology exhibited mild pathological alteration with different injures between treated groups. Splenic white pulp accompanied by ill deformed which reflected the reduction of white pulp zones, thickened vasculature in the splenic net work, fibrous trabeculae become prominent feature, where splenic red pulp occupied large areas of the splenic network with predominant edema and megakaryocytes. On the other hand, the effect of tramadol and/or APAP induced DNA alterations of hepatocytes in dose dependent pattern as elucidated by dendrogramatic analysis. Liver histopathological changes of treated groups included vacuolated hepatocytes, dilated sinusoid with proliferated Kupffer cells; atrophied hepatocytes with nuclei reduced in size and darkly stained. Many areas of hepatocytes showed loss of architecture, congested central vein, expanded portal area with edema and inflammatory reaction. It could be concluded that the effect of tramadol/APAP induced anti-inflammatory cytokines than tramadol and APAP alone. Tramadol and/or APAP may display severe pathological consequences of hepatocytes. These hepatic lesions may be caused impairment of the liver function


Subject(s)
Male , Animals, Laboratory , Acetaminophen/adverse effects , Pain/drug therapy , Phagocytosis/drug effects , Cytokines , Drug Combinations/adverse effects , DNA Fragmentation/drug effects , Liver/pathology , Histology , Rats , Models, Animal
9.
Article in English | WPRIM | ID: wpr-203590

ABSTRACT

The action mode of 4,4'-diaminodiphenylsulfone (DDS) is still under debate, although it has long been used in treatment of several dermatologic diseases including Hansen's disease. In this study, we tested the effect of DDS as an antioxidant on paraquat-induced oxidative stress in non-phagocytic human diploid fibroblasts (HDFs). Overall, preincubation of HDFs with DDS prevented the oxidative stress and the resulting cytotoxic damages caused by paraquat in these cells. The specific effects of DDS in paraquat-treated HDFs are summarized as follows: a) reducing the expression of NADPH oxidase 4 (NOX4) by inhibiting paraquat-induced activation of PKC; b) inhibiting paraquat-induced decreases in mitochondrial complex protein levels as well as in membrane potentials; c) consequently, inhibiting the generation of cytosolic and mitochondrial superoxide anions. Taken together, these findings suggest that DDS would suppress the radical generation in non-phagocytic HDFs during oxidative stress, and that DDS might have the extended potential to be used further in prevention of other oxidative stress-related pathologies.


Subject(s)
Biphenyl Compounds/metabolism , Cell Death/drug effects , Cell Survival/drug effects , Dapsone/pharmacology , Diploidy , Enzyme Activation/drug effects , Fibroblasts/cytology , Free Radical Scavengers/metabolism , Gene Expression Regulation, Enzymologic/drug effects , Humans , Male , Mitochondria/drug effects , NADPH Oxidases/genetics , Paraquat/toxicity , Phagocytosis/drug effects , Picrates/metabolism , Protein Kinase C/metabolism , RNA, Messenger/genetics , Reactive Oxygen Species/metabolism , Superoxides/metabolism
10.
Article in English | WPRIM | ID: wpr-197698

ABSTRACT

The objective of this study was to explore the immunomodulatory effects of betulinic acid (BA) extracted from the bark of white birch on mice. Female mice were orally administered BA for 14 days in doses of 0, 0.25, 0.5, and 1 mg/kg body weight. We found that BA significantly enhanced the thymus and spleen indices, and stimulated lymphocyte proliferation induced by Concanavalin A and lipopolysaccharide as shown by MTT assay. Flow cytometry revealed that BA increased the percentage of CD4+ cells in thymus as well as the percentage of CD19+ and the ratios of CD4+/CD8+ in spleen. BA increased the number of plaque-forming cell and macrophage phagocytic activity as indicated by a neutral red dye uptake assay, and the peritoneal macrophages levels of TNF-alpha were also increased. In contrast, serum levels of IgG and IgM and serum concentrations of IL-2 and IL-6 were significantly decreased in BA-treated mice compared to the control as assayed by haemagglutination tests and ELISA, respectively. Taken together, these results suggest that BA enhances mouse cellular immunity, humoral immunity, and activity of macrophages. Thus, BA is a potential immune stimulator and may strengthen the immune response of its host.


Subject(s)
Adaptive Immunity/drug effects , Animals , Betula/chemistry , Cell Proliferation/drug effects , Cytokines/blood , Female , Immunity, Innate/drug effects , Immunologic Factors/pharmacology , Macrophages/drug effects , Mice , Phagocytosis/drug effects , Random Allocation , Spleen/cytology , Thymus Gland/cytology , Triterpenes/pharmacology
11.
Braz. j. med. biol. res ; 42(10): 912-917, Oct. 2009. ilus, tab
Article in English | LILACS | ID: lil-526193

ABSTRACT

Cyhalothrin, a pyrethroid insecticide, induces stress-like symptoms, increases c-fos immunoreactivity in the paraventricular nucleus of the hypothalamus, and decreases innate immune responses in laboratory animals. Macrophages are key elements in cellular immune responses and operate at the tumor-host interface. This study investigated the relationship among cyhalothrin effects on Ehrlich tumor growth, serum corticosterone levels and peritoneal macrophage activity in mice. Three experiments were done with 10 experimental (single gavage administration of 3.0 mg/kg cyhalothrin daily for 7 days) and 10 control (single gavage administration of 1.0 mL/kg vehicle of cyhalothrin preparation daily for 7 days) isogenic BALB/c mice in each experiment. Cyhalothrin i) increased Ehrlich ascitic tumor growth after ip administration of 5.0 x 106 tumor cells, i.e., ascitic fluid volume (control = 1.97 ± 0.39 mL and experimental = 2.71 ± 0.92 mL; P < 0.05), concentration of tumor cells/mL in the ascitic fluid (control = 111.95 ± 16.73 x 106 and experimental = 144.60 ± 33.18 x 106; P < 0.05), and total number of tumor cells in the ascitic fluid (control = 226.91 ± 43.22 x 106 and experimental = 349.40 ± 106.38 x 106; P < 0.05); ii) increased serum corticosterone levels (control = 200.0 ± 48.3 ng/mL and experimental = 420.0 ± 75.5 ng/mL; P < 0.05), and iii) decreased the intensity of macrophage phagocytosis (control = 132.3 ± 19.7 and experimental = 116.2 ± 4.6; P < 0.05) and oxidative burst (control = 173.7 ± 40.8 and experimental= 99.58 ± 41.7; P < 0.05) in vitro in the presence of Staphylococcus aureus. These data provide evidence that cyhalothrin simultaneously alters host resistance to Ehrlich tumor growth, hypothalamic-pituitary-adrenocortical (HPA) axis function, and peritoneal macrophage activity. The results are discussed in terms of data suggesting a link between stress, HPA axis activation and resistance to tumor growth.


Subject(s)
Animals , Male , Mice , Carcinoma, Ehrlich Tumor/pathology , Insecticides/pharmacology , Macrophages, Peritoneal/drug effects , Nitriles/pharmacology , Phagocytosis/drug effects , Pyrethrins/pharmacology , Carcinoma, Ehrlich Tumor/blood , Corticosterone/blood , Hypothalamo-Hypophyseal System/drug effects , Mice, Inbred BALB C , Tumor Cells, Cultured
12.
Braz. j. med. biol. res ; 42(7): 599-605, July 2009. graf
Article in English | LILACS | ID: lil-517795

ABSTRACT

To determine the effects of saturated and unsaturated fatty acids in phosphatidylcholine (PC) on macrophage activity, peritoneal lavage cells were cultured in the presence of phosphatidylcholine rich in saturated or unsaturated fatty acids (sat PC and unsatPC, respectively), both used at concentrations of 32 and 64 ìM. The treatment of peritoneal macrophages with 64 ìM unsat PC increased the production of hydrogen peroxide by 48.3% compared to control (148.3 ± 16.3 vs 100.0 ± 1.8%, N = 15), and both doses of unsat PC increased adhesion capacity by nearly 50%. Moreover, 64 ìM unsat PC decreased neutral red uptake by lysosomes by 32.5% compared to the untreated group (67.5 ± 6.8 vs 100.0 ± 5.5%, N = 15), while both 32 and 64 ìM unsat PC decreased the production of lipopolysaccharide-elicited nitric oxide by 30.4% (13.5 ± 2.6 vs 19.4 ± 2.5 ìM) and 46.4% (10.4 ± 3.1 vs 19.4 ± 2.5 ìM), respectively. Unsat PC did not affect anion production in non-stimulated cells or phagocytosis of unopsonized zymosan particles. A different result pattern was obtained for macrophages treated with sat PC. Phorbol 12-miristate 13-acetate-elicited superoxide production and neutral red uptake were decreased by nearly 25% by 32 and 64 ìM sat PC, respectively. Sat PC did not affect nitric oxide or hydrogen peroxide production, adhesion capacity or zymosan phagocytosis. Thus, PC modifies macrophage activity, but this effect depends on cell activation state, fatty acid saturation and esterification to PC molecule and PC concentration. Taken together, these results indicate that the fatty acid moiety of PC modulates macrophage activity and, consequently, is likely to affect immune system regulation in vivo.


Subject(s)
Animals , Male , Rats , Linoleic Acids/pharmacology , Macrophages, Peritoneal/drug effects , Phagocytosis/drug effects , Phosphatidylcholines/pharmacology , Cell Adhesion/drug effects , Cell Adhesion/physiology , Hydrogen Peroxide/metabolism , Macrophages, Peritoneal/physiology , Nitric Oxide Synthase/metabolism , Nitric Oxide/metabolism , Phagocytosis/physiology , Rats, Wistar , Reactive Oxygen Species/metabolism
13.
Rev. biol. trop ; 56(3): 1063-1073, sep. 2008. ilus, graf
Article in Spanish | LILACS | ID: lil-637847

ABSTRACT

Sesquiterpene lactones of the plants Viguiera sylvatica and Decachaeta thieleana (Asteraceae) modulate nitric oxide production and phagocytosis of macrophages cell line RAW. Different species of the Asteraceae family are a potential source of sesquiterpene lactones with anti-inflammatory properties. Macrophages play a central role in the regulation of immune responses. In the present study, the in vitro effect of two sesquiterpene lactones, a millerenolide and a thieleanin, was assessed by measuring the production of nitric oxide (NO) by cell line RAW (murine macrophages) using the Griess reagent. Additionally, the effect of these sesquiterpene lactones on phagocytic capacity of latex particles and the reduction of nitroblue tetrazolium (NBT) were evaluated microscopically. Treatment of macrophages with >2.5 µg/ml of both sesquiterpene lactones, reduced the production of NO. A decreased number of macrophages able to reduce NBT were observed when these cells were treated with 3 µg/ml of millerenolide or 7.5 µg/ml of thieleanin. Treatment of macrophages with 4 µg/ml of millerenolide or 7.5 µg/ml of thieleanin, reduced the phagocytic capacity of macrophages. Cytotoxic effects on the macrophages were only observed when the concentration was increased to 8 µg/ml of millerenolide or 25 µg/ml of thieleanin. Our results suggest that these sesquiterpene lactones could be useful compounds in the elaboration of anti-inflammatory drugs. Rev. Biol. Trop. 56 (3): 1063-1073. Epub 2008 September 30.


Las plantas de la familia Asteraceae son una fuente potencial de lactonas sesquiterpénicas con propiedades antiinflamatorias. Los macrófagos son células que desempeñan una función central en la regulación de la respuesta inmune. Este estudio evaluó el efecto in vitro de dos lactonas sesquiterpénicas, un millerenólido y thieleanina, sobre la producción de óxido nítrico (NO) en una línea celular de macrófagos de ratón (RAW), utilizando el reactivo de Griess. Además, se estudió el efecto sobre la capacidad fagocítica de RAW, evaluando al microscopio la fagocitosis de partículas inertes de látex y la reducción de nitroazul de tetrazolio (NBT). Se observó que los macrófagos tratados con lactona sesquiterpénica (>2.5 µg/ml) disminuyeron la producción de NO. Además, se observó una disminución de la cantidad de macrófagos capaces de reducir NBT, después que fueron tratados con millerenólido (3 µg/ml) o thieleanina (7.5 µg/ ml). Por otro lado, la exposición a 4 µg/ml de millerenólido ó 7.5 µg/ml de thieleanina redujo la cantidad promedio de partículas de látex fagocitadas. No se observaron diferencias entre tratamientos y control en cuanto al porcentaje de células fagocíticas. Sólo se observaron efectos citotóxicos sobre los macrófagos, cuando la concentración de millerenólido se incrementó a 8 µg/ml o la de thieleanina se aumentó a 25 µg/ml. Estos resultados sugieren que el millerenólido y la thieleanina podrían ser compuestos útiles en la elaboración de drogas antiinflamatorias.


Subject(s)
Animals , Mice , Asteraceae/chemistry , Lactones/pharmacology , Macrophages/drug effects , Nitric Oxide/biosynthesis , Phagocytosis/drug effects , Sesquiterpenes/pharmacology , Cell Line , Lactones/chemistry , Lactones/isolation & purification , Macrophages/physiology , Sesquiterpenes/chemistry , Sesquiterpenes/isolation & purification
14.
Mem. Inst. Oswaldo Cruz ; 103(3): 244-250, May 2008. ilus, graf
Article in English | LILACS | ID: lil-485211

ABSTRACT

A polysaccharide-rich fraction (ATF) of medicinal mushroom Agaricus brasiliensis was evaluated on the candidacidal activity, H2O2 and nitric oxide (NO) production, and expression of mannose receptors by murine peritoneal macrophages. Mice received three intraperitoneal (i.p.) injections of ATF and after 48 h their peritoneal resident macrophages were assayed against Candida albicans yeast forms. The treatment increased fungicidal activity and it was associated with higher levels of H2O2, whereas NO production was not affected. We also found that the treatment enhances mannose receptor expression by peritoneal macrophages, which are involved in the attachment and phagocytosis of non-opsonized microorganisms. Treatment of animals with ATF was able to enhance the clearance of C. albicans during the first 6 h after the experimental i.p. infection. Our results suggest that this extract can increase host resistance against some infectious agents through the stimulation of microbicidal activity of macrophages.


Subject(s)
Animals , Male , Mice , Agaricus/chemistry , Candida albicans/immunology , Macrophages, Peritoneal/immunology , Polysaccharides/pharmacology , Candida albicans/drug effects , Hydrogen Peroxide/immunology , Lectins, C-Type/immunology , Mice, Inbred BALB C , Macrophages, Peritoneal/drug effects , Macrophages, Peritoneal/microbiology , Mannose-Binding Lectins/immunology , Nitric Oxide/biosynthesis , Phagocytosis/drug effects , Polysaccharides/isolation & purification , Receptors, Cell Surface/immunology
15.
Indian J Med Microbiol ; 2008 Jan-Mar; 26(1): 25-8
Article in English | IMSEAR | ID: sea-53651

ABSTRACT

PURPOSE: To eliminate pathogenic bacteria, the host presents conditions that are stressful for bacteria. Oxidative stress arises when the concentration of pro-oxidants like hydrogen peroxide (H2O2 ) and superoxide anion increases to a level over the basal defence capacity of the cell. In the present study, we studied the effect of oxidative stress on the production of certain virulence factors by Escherichia coli . METHODS: E. coli was exposed to oxidative stress by growing in the presence of different concentrations of H2O2 . The effect of oxidative stress on the expression of surface hydrophobicity, adherence, haemolysin production, serum resistance and phagocytosis was studied. RESULTS: Oxidative stress caused a significant decrease in the expression of all the virulence factors of E. coli . CONCLUSIONS: Synthesis of virulence factors can be significantly altered by oxidative stress and such changes may affect the pathogenicity of E. coli.


Subject(s)
Anti-Bacterial Agents/pharmacology , Bacterial Adhesion/drug effects , Escherichia coli/drug effects , Escherichia coli Proteins/biosynthesis , Hemolysin Proteins/biosynthesis , Humans , Hydrogen Peroxide/pharmacology , Hydrophobic and Hydrophilic Interactions , Phagocytosis/drug effects , Virulence/drug effects , Virulence Factors/biosynthesis
16.
Rev. méd. Chile ; 136(1): 64-72, ene. 2008. ilus, graf
Article in Spanish | LILACS | ID: lil-483221

ABSTRACT

Background: Natural products are used in the production of therapeutic drugs due to their wide diversity and excellent adaptability to biological structures. Sesquiterpene ¡aciones are the active constituents of several plants from the Asteraceae family. Aim: To assess the in vitro effect of a sesquiterpene lactone (millerenolide). Material and methods: The drug effect was assessed measuring the proliferation of lymphocytes using the 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonylJ-2H-tetrazolium hydroxide (XTT) technique. Changes on the cell cycle were analyzed on a FACSort flow cytometer The effect of millerenolide on the production of nitric oxide (NO) by macrophages was evaluated using the Griess reagent. Additionally, phagocytosis of latex particles and nitroblue tetrazolium (NBT) reduction by macrophages were evaluated microscopically. Results: Treatment of human peripheral blood mononuclear cells (PBMC) with millerenolide decreases the proliferation of lymphocytes, decreases the percentage of cells in S, and G2/Mphases, and increases the proportion of cells in GO/Gl phase. Treatment of macrophages with millerenolide, reduces the production of NO, the phagocytic capacity and the number of cells able to reduce NBT. Cytotoxic effects of the lactone on human PBMC were only observed when the concentration was increased to 6 fig/ml. Conclusions: Millerenolide could be considered as a potential therapeutic agent with immunosuppressiveproperties.


Subject(s)
Humans , In Vitro Techniques , Immunosuppressive Agents/pharmacology , Lactones/pharmacology , Leukocytes, Mononuclear/drug effects , Lymphocyte Activation/drug effects , Nitric Oxide/biosynthesis , Sesquiterpenes/pharmacology , Analysis of Variance , Asteraceae/chemistry , Cytotoxicity Tests, Immunologic , Lactones/chemistry , Lactones/toxicity , Leukocytes, Mononuclear/physiology , Lymphocyte Activation/physiology , Phagocytosis/drug effects , Phagocytosis/physiology , Plant Extracts/chemistry , Plant Extracts/toxicity , Sesquiterpenes/chemistry , Sesquiterpenes/toxicity
17.
Indian J Physiol Pharmacol ; 2007 Oct-Dec; 51(4): 387-94
Article in English | IMSEAR | ID: sea-107186

ABSTRACT

Centchroman (Ormeloxifene) is a nonsteroidal selective estrogen receptor modulator that is used as once a week oral contraceptive agent. The effect of centchroman on the immune system was evaluated by using different experimental models such as carbon clearance test, cyclophosphamide induced neutropenia, neutrophil adhesion test, effect on serum immunoglobulins, mice lethality test and indirect haemagglutination test. The first three models namely carbon clearance test, cyclophosphamide induced neutropenia and neutrophil adhesion test were used to study cell mediated immunity while the latter three models were used to see the effect on humoral immunity. Centchroman was administered orally at a dose of 5 mg/kg and levamisole (2.5 mg/kg/ p.o) was used as standard drug. Centchroman significantly increased the levels of serum immunoglobulins and also prevented the mortality induced by bovine Pasteurella multocida in mice. It also increased significantly the circulating antibody litre in indirect haemagglunation test. However, it did not show any significant effect on phagocytic index in carbon clearance assay and nor did influence the adhesion of neutrophils in the neutrophil adhesion test. Centchroman was also not effective in preventing the cyclophosphamde induced neutropenia. Hence, it was concluded that centchroman increases humoral immunity with no significant effect on cell mediated immunity.


Subject(s)
Animals , Antibody Formation/drug effects , Cell Adhesion , Centchroman/pharmacology , Contraceptives, Postcoital, Synthetic/pharmacology , Cyclophosphamide/pharmacology , Female , Hemagglutination Tests , Immunity, Cellular/drug effects , Immunoglobulins/blood , Mice , Neutrophils/drug effects , Phagocytosis/drug effects , Rats , Rats, Wistar , Selective Estrogen Receptor Modulators/pharmacology
18.
Pakistan Journal of Pharmaceutical Sciences. 2007; 20 (3): 175-179
in English | IMEMR | ID: emr-134955

ABSTRACT

Effects of Saussurea lappa root extracts prepared in ethanol according to the homeopathic principles were ssessed on leukocyte phagocytic activity, lymphocyte transformation and mitogen-induced interferon-gamma [INF-gamma] in the cultures of peripheral blood mononuclear cells of goats [PBMC] in vitro. Leukocyte phagocytic activity was measured by flow cytometry, lymphocyte proliferation by MTT and IFN-gamma level in cell culture supernatants was determined by ELISA. The results obtained demonstrated that all tests dilutions [D4, D6, D8] of Saussurea lappa in ethanol have exerted a stimulating effect on leukocyte phagocytic activity in dose-dependent manner. A 10 micro liters dose of Saussurea lappa of each dilution markedly enhanced phagocytic activity, while other does tested made only a feeble stimulating effect. The increase with 10 micro liter dose were found significantly [p<0.01] different between each dilution, maximal stimulationwas observed by D8 dilution. Different does [10 micro liters, 2 micro liters, 0.5 micro liters] of all test dilutions [D4, D6, D8] of Saussurea lappa in sterile 0.9% NaCl solution inhibited lymphocyte proliferation. Maximal inhibitory effect was observed with the 2 micro liter dose. Similarly, Saussurea lappa suppressed the secretion of IFN-gamma by mitoren-activated [PHA; 2.5 micro g/ml] of peripheral mononuclear cells in dose-dependent manner. In conclusion these findings suggest that enhanced leukocyte phagocytic activity may be helpful to clear the soluble immune complexes produced during a sustained immune response against self antigens which causes chronic inflammatory injury of tissue. On the other hand, inhibition of lymphocyte proliferation and IFN-gamma by Saussurea lappa may contribute to suppress immune-mediated inflammatory reactions possibly through a cell-mediated cytokine pathway. Thus it is conceivable that ethanolic extracts of Saussurea lappa roots in homeopathetic dilutions may be considered as a potential candidate for therapeutic support in autoimmune and chronic inflammatory disorders


Subject(s)
Animals , Plant Roots , Leukocytes/drug effects , Phagocytosis/drug effects , Lymphocytes/drug effects , Interferon-gamma/drug effects , Plant Extracts , Ethanol , Goats
19.
Article in English | WPRIM | ID: wpr-191499

ABSTRACT

Receptor activator of NFkappaB ligand (RANKL) is known as a key regulator of osteoclastogenesis. However, the fact that fibroblasts and periodontal ligament cells express RANKL in response to bacterial substances, suggests that RANKL may have evolved as a part of the immunity to infection. As RANKL increases the survival and activity of dendritic cells, it may have similar effects on macrophages. To address this issue, we studied the effect of RANKL on various functions of macrophages using mouse bone marrow derived macrophages. RANKL enhanced the survival of macrophages and up-regulated the expression of CD86. RANKL-treated macrophages showed increased allogeneic T cell activation and phagocytic activity compared to control cells. In addition, RANKL increased the expression of TNFalpha, MCP-1, and IL-6 but not of IL-10, IL-12, IFN-gamma, and iNOS. Collectively, RANKL augmented the activity of macrophages especially as antigen presenting cells, suggesting its new role in immune regulation.


Subject(s)
Animals , Antigen-Presenting Cells/cytology , B7-2 Antigen/metabolism , Carrier Proteins/pharmacology , Cell Death/drug effects , Cell Survival/drug effects , Cells, Cultured , Cytokines/metabolism , Flow Cytometry , Histocompatibility Antigens Class II/metabolism , Inflammation Mediators , Interferon-gamma/pharmacology , Lipopolysaccharides/pharmacology , Macrophages/cytology , Membrane Glycoproteins/pharmacology , Mice , Mice, Inbred C57BL , Mice, Inbred ICR , Nitric Oxide Synthase Type II/metabolism , Phagocytosis/drug effects , T-Lymphocytes/immunology , Up-Regulation/drug effects
20.
Article in English | WPRIM | ID: wpr-37855

ABSTRACT

Dendritic cells (DCs) play a key role in activating the immune response against invading pathogens as well as dying cells or tumors. Although the immune response can be initiated by the phagocytic activity by DCs, the molecular mechanism involved in this process has not been fully investigated. Trp-Lys-Tyr-Met-Val-Met-NH2 (WKYMVM) stimulates the activation of phospholipase D (PLD) via Ca2+ increase and protein kinase C activation in mouse DC cell line, DC2.4. WKYMVM stimulates the phagocytic activity, which is inhibited in the presence of N-butanol but not t-butanol in DC2.4 cells. Furthermore, the addition of phosphatidic acid, an enzymatic product of PLD activity, enhanced the phagocytic activity in DC2.4 cells. Since at least two of formyl peptide receptor (FPR) family (FPR1 and FPR2) are expressed in DC2.4 as well as in mouse bone marrow-derived dendritic cells, this study suggests that the activation of FPR family by WKYMVM stimulates the PLD activity resulting in phagocytic activity in DC2.4 cells.


Subject(s)
1-Butanol/pharmacology , Animals , Bone Marrow Cells/cytology , Calcium Signaling/drug effects , Cell Death/immunology , Cell Line , Communicable Diseases/immunology , Dendritic Cells/immunology , Mice , Neoplasms/immunology , Oligopeptides/pharmacology , Phagocytosis/drug effects , Phosphatidic Acids/pharmacology , Phospholipase D/metabolism , Receptors, Formyl Peptide/metabolism , tert-Butyl Alcohol/pharmacology
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