ABSTRACT
Galectin-3 (Gal-3) controls intercellular and cell-extracellular matrix interactions during immunological responses. In chronic inflammation, Gal-3 is associated with fibrotic events, regulates B cell differentiation and delays lupus progression. Gal-3 deficient mice (Lgals3-/-) have intense germinal center formation and atypical plasma cell generation correlated to high levels IgG, IgE, and IgA. Here, we used pristane (2,6,10,14-tetramethylpentadecane) to induce lupus-like syndrome in Lgals3-/- and Lgals3+/+ BALB/c mice. Mesentery and peritoneal cells were monitored because promptly react to pristane injected in the peritoneal cavity. For the first time, mesenteric tissues have been associated to the pathogenesis of experimental lupus-like syndrome. In Lgals3+/+ pristane-induced mice, mesentery was hallmarked by intense fibrogranulomatous reaction restricted to submesothelial regions and organized niches containing macrophages and B lymphocytes and plasma cells. In contrast, Lgals3-/- pristane-treated mice had diffuse mesenteric fibrosis affecting submesothelium and peripheral tissues, atypical M1/M2 macrophage polarization and significant DLL1+ cells expansion, suggesting possible involvement of Notch/Delta pathways in the disease. Early inflammatory reaction to pristane was characterized by significant disturbances on monocyte recruitment, macrophage differentiation and dendritic cell (DC) responses in the peritoneal cavity of pristane-induced Lgals3-/- mice. A correlative analysis showed that mesenteric damages in the absence of Gal-3 were directly associated with severe portal inflammation and hepatitis. In conclusion, it has suggested that Gal-3 orchestrates histological organization in the mesentery and prevents lupoid hepatitis in experimental lupus-like syndrome by controlling macrophage polarization, Notch signaling pathways and DC differentiation in mesenteric structures.
Subject(s)
Galectin 3/metabolism , Hepatitis/immunology , Lupus Erythematosus, Systemic/immunology , Macrophages, Peritoneal/immunology , Mesentery/pathology , Animals , Disease Models, Animal , Female , Fibrosis , Galectin 3/genetics , Hepatitis/pathology , Humans , Injections, Intraperitoneal , Liver/immunology , Liver/pathology , Lupus Erythematosus, Systemic/chemically induced , Lupus Erythematosus, Systemic/complications , Mesentery/cytology , Mesentery/immunology , Mice , Mice, Knockout , Terpenes/administration & dosage , Terpenes/immunologyABSTRACT
Envenomation caused by spiders Loxosceles induce intense dermonecrosis at the bite site and systemic disease. In this work we described the hyaluronidase and collagenase activities in vitro of the Loxosceles intermedia venom, but no phospholipase A2 activity. In vivo, we evaluated the effect of L. intermedia venom used different strain of mice, C57BL/6, BALB/c and Swiss. All mice developed paw edema after venom injection, persistent for 24 h in BALB/c and C57BL/6 mice. Histopathological analysis of the skin after venom injection revealed vascular congestion in Swiss mice and an inflammatory reaction in BALB/c and C57BL/6 mice. The mobilization of inflammatory cells from bone marrow, spleen and blood was investigated. Typical innate immune response with mobilization of myeloid cells and cytotoxic CD8 T lymphocytes was observed in C57BL/6 mice. In contrast, typical acquired/humoral immune response was observed in BALB/c mice, with preferential involvement of conventional B lymphocytes and CD4 T helper cells. The skin inflammation associated to mobilization of inflammatory cells indicated that mice models are strongly recommended to investigate specific cell types involved with immune response to the envenomation and mechanisms to inhibit skin lesions.
Subject(s)
Inflammation/pathology , Necrosis/chemically induced , Necrosis/pathology , Phosphoric Diester Hydrolases/toxicity , Skin/pathology , Spider Venoms/toxicity , Spiders/chemistry , Analysis of Variance , Animals , CD8-Positive T-Lymphocytes/immunology , Collagenases/metabolism , Flow Cytometry , Hyaluronoglucosaminidase/metabolism , Mice , Mice, Mutant Strains , Phospholipases/metabolism , Phosphoric Diester Hydrolases/immunology , Species Specificity , Sphingomyelin Phosphodiesterase/metabolism , Spider Venoms/immunologyABSTRACT
We studied the effect of pulsed ultrasound therapy (UST) and antibothropic polyvalent antivenom (PAV) on the regeneration of mouse extensor digitorum longus muscle following damage by Bothrops jararacussu venom. Animals (Swiss male and female mice weighing 25.0 ± 5.0 g; 5 animals per group) received a perimuscular injection of venom (1 mg/kg) and treatment with UST was started 1 h later (1 min/day, 3 MHz, 0.3 W/cm², pulsed mode). Three and 28 days after injection, muscles were dissected and processed for light microscopy. The venom caused complete degeneration of muscle fibers. UST alone and combined with PAV (1.0 mL/kg) partially protected these fibers, whereas muscles receiving no treatment showed disorganized fascicules and fibers with reduced diameter. Treatment with UST and PAV decreased the effects of the venom on creatine kinase content and motor activity (approximately 75 and 48%, respectively). Sonication of the venom solution immediately before application decreased the in vivo and ex vivo myotoxic activities (approximately 60 and 50%, respectively). The present data show that UST counteracts some effects of B. jararacussu venom, causing structural and functional improvement of the regenerated muscle after venom injury.
Subject(s)
Animals , Female , Male , Mice , Antivenins/pharmacology , Bothrops , Crotalid Venoms/poisoning , Muscle, Skeletal/drug effects , Snake Bites/therapy , Ultrasonic Therapy/methods , Creatine Kinase/metabolism , Crotalid Venoms/administration & dosage , Edema/chemically induced , Immunologic Factors/immunology , Motor Activity/drug effects , Muscle, Skeletal/enzymology , Muscle, Skeletal/pathology , Necrosis , Rotarod Performance Test , Regeneration/drug effects , Snake Bites/complicationsABSTRACT
We studied the effect of pulsed ultrasound therapy (UST) and antibothropic polyvalent antivenom (PAV) on the regeneration of mouse extensor digitorum longus muscle following damage by Bothrops jararacussu venom. Animals (Swiss male and female mice weighing 25.0 ± 5.0 g; 5 animals per group) received a perimuscular injection of venom (1 mg/kg) and treatment with UST was started 1 h later (1 min/day, 3 MHz, 0.3 W/cm(2), pulsed mode). Three and 28 days after injection, muscles were dissected and processed for light microscopy. The venom caused complete degeneration of muscle fibers. UST alone and combined with PAV (1.0 mL/kg) partially protected these fibers, whereas muscles receiving no treatment showed disorganized fascicules and fibers with reduced diameter. Treatment with UST and PAV decreased the effects of the venom on creatine kinase content and motor activity (approximately 75 and 48%, respectively). Sonication of the venom solution immediately before application decreased the in vivo and ex vivo myotoxic activities (approximately 60 and 50%, respectively). The present data show that UST counteracts some effects of B. jararacussu venom, causing structural and functional improvement of the regenerated muscle after venom injury.
Subject(s)
Antivenins/pharmacology , Bothrops , Crotalid Venoms/poisoning , Muscle, Skeletal/drug effects , Snake Bites/therapy , Ultrasonic Therapy/methods , Animals , Creatine Kinase/metabolism , Crotalid Venoms/administration & dosage , Edema/chemically induced , Female , Immunologic Factors/immunology , Male , Mice , Motor Activity/drug effects , Muscle, Skeletal/enzymology , Muscle, Skeletal/pathology , Necrosis , Regeneration/drug effects , Rotarod Performance Test , Snake Bites/complicationsABSTRACT
The effect of dose and volume of a perimuscular injection of Bothrops jararacussu venom on myonecrosis of skeletal muscle was studied in mice. An increase of the venom dose (0.25 to 2.0 micro g/g) at a given volume (50 micro l) resulted in an increase in plasma creatine kinase (CK) levels 2 h after injection. Plasma CK activity increased from the basal level of 129.27 +/- 11.83 (N = 20) to 2392.80 +/- 709.43 IU/l (N = 4) for the 1.0 micro g/g dose. Histological analysis of extensor digitorum longus muscle 4 h after injection showed lesion of peripheral muscle fibers, disorganization of the bundles or the complete degeneration of muscle fibers. These lesions were more extensive when higher doses were injected. Furthermore, an increase in volume (12.5 to 100 micro l) by dilution of a given dose (0.5 micro g/g) also increased plasma CK levels from 482.31 +/- 122.79 to 919.07 +/- 133.33 IU/l (N = 4), respectively. These results indicate that care should be taken to standardize volumes and sites of venom injections.
Subject(s)
Bothrops , Creatine Kinase/blood , Crotalid Venoms/administration & dosage , Muscle, Skeletal/drug effects , Animals , Crotalid Venoms/pharmacology , Dose-Response Relationship, Drug , Injections, Intramuscular , Mice , Muscle, Skeletal/pathology , NecrosisABSTRACT
The effect of dose and volume of a perimuscular injection of Bothrops jararacussu venom on myonecrosis of skeletal muscle was studied in mice. An increase of the venom dose (0.25 to 2.0 æg/g) at a given volume (50 æl) resulted in an increase in plasma creatine kinase (CK) levels 2 h after injection. Plasma CK activity increased from the basal level of 129.27 ± 11.83 (N = 20) to 2392.80 ± 709.43 IU/l (N = 4) for the 1.0 æg/g dose. Histological analysis of extensor digitorum longus muscle 4 h after injection showed lesion of peripheral muscle fibers, disorganization of the bundles or the complete degeneration of muscle fibers. These lesions were more extensive when higher doses were injected. Furthermore, an increase in volume (12.5 to 100 æl) by dilution of a given dose (0.5 æg/g) also increased plasma CK levels from 482.31 ± 122.79 to 919.07 ± 133.33 IU/l (N = 4), respectively. These results indicate that care should be taken to standardize volumes and sites of venom injections
Subject(s)
Animals , Mice , Bothrops , Creatine Kinase , Crotalid Venoms , Muscle, Skeletal , Crotalid Venoms , Dose-Response Relationship, Drug , Muscle, SkeletalABSTRACT
We examined the effect of treatment with heparin and polyvalent antivenom on mice muscle Extensor digitorum longus (EDL) regeneration, after damage induced by injection of Bothrops jararacussu crude venom over the muscle of the right posterior limb. The mice were separated into groups and each group received treatment, by intravenous route with either high molecular weight heparin (H), low molecular weight heparin (LMWH), polyvalent antivenom (PAV) or with the combination of PAV plus H or PAV plus LMWH at 15 minutes and 4 hours after the injection of the venom. Myotoxicity was measured by the increase in plasma creatine kinase (CK) activity at two hours after the injection of the venom. The histological changes in EDL at 1, 3, 7 and 21 days after the injection of the venom were analyzed by light microscopy. In each group the normal and regenerated muscle fibers were quantified using Scion Image computer program. We also evaluated in vitro, the influence of these substances in the proteolytic and phospholipase activities of the venom. Heparins decreased the proteolytic activity of the venom but did not affect its phospholipase activity. However the PAV antagonized both activities. PAV and its combinations showed antimyotoxic activity, according to the magnitude of CK plasma levels. At 21 days the regeneration was observed in all animals, also in those that received only the venom. All treatments, except LMWH, promote a significant increase in the number of muscle fibers.
Subject(s)
Bothrops , Crotalid Venoms/pharmacology , Heparin/pharmacology , Muscle, Skeletal/injuries , Animals , Creatine Kinase/metabolism , Crotalid Venoms/immunology , Heparin, Low-Molecular-Weight/pharmacology , Horses , Mice , Muscle, Skeletal/drug effects , Muscle, Skeletal/pathology , Phospholipases A/metabolismABSTRACT
An acidic phospholipase A2 isolated from Lachesis muta snake venom denoted LM-PLA2, showed neither toxic nor anticoagulant activities in contrast to a potent inhibitory effect of collagen-induced platelet aggregation [Fuly, A.L., Machado. O.L.T., Alves, E.W. and Carlini, C.R., 1997. Thromb. Haemost 78, 1372-1380.]. Now, the myotoxicity induced by LM-PLA2 was investigated by using both in vivo and in vitro experiments. LM-PLA2 induced in vitro a dose- and time-dependent release of creatine-kinase (CK) from mouse Extensor Digitorium Longus (EDL) muscles and also increased the plasma CK activity in treated animals. Histopathological studies confirm myonecrosis of mouse skeletal muscles as a major effect. Edema could also be seen in muscle tissue. The amino-terminal sequence of LM-PLA2 (previously reported) indicates an aspartic acid residue located at position 49, together with other conserved amino acids present in the Asp-49 phospholipases, such as Tyr-28, Gly-30, Gly-32, His-48. Chemical modification of the protein moiety was also performed. Histidine alkylation with p-bromophenacyl bromide and lysine acetylation with acetic anhydride, abolished both indirect hemolytic and myotoxic activities of LM-PLA2. On the other hand, contrarily to what has been observed with several basic myotoxic phospholipases, the myotoxic effect induced by LM-PLA2 was not abolished by heparin.
