RESUMEN
A 56-day research was performed to examine the influence of graded levels (0 (control), 0.5, 1 and 2%) of Fern (Adiantum capillus-veneris) leaves powder (FLP) in diet on immune competence and growth of common carp (Cyprinus carpio, initial weight = 20 g). The serum total immunoglobulins content and lysozyme activity in the 1 and 2% FLP groups remarkably increased compared to the other groups (P < 0.05). The skin mucosal lysozyme activity enhanced with increasing dietary FLP level in a dose-response manner. Fish fed on the FLP-supplemented diets had higher skin-mucosal superoxide dismutase activity than the control (P < 0.05). However, serum antioxidant enzymes were not affected by dietary fern (P > 0.05). The serum bactericidal activity against human and fish pathogens increased with enhancing the FLP level in diet against Staphylococcus aureus, Escherichia coli (EHEC ATCC 43895), Escherichia coli (CI), Pseudomonas aeruginosa, Klebsiella pneumonia and Aeromonas hydrophila. The serum antibacterial activity against Yersinia ruckeri in the 2% FLP group was higher than the other treatments. Furthermore, the serum bactericidal activity against P. aeruginosa (ATCC 27853) only observed in fish fed on the 1 and 2% FLP-supplemented diets. The skin mucosal bactericidal activity and inhibitory effects increased with enhancing the FLP level in diet against E. coli, K. pneumonia, Y. ruckeri and A. hydrophila in a dose response manner. Moreover, the skin mucosal bactericidal activity against S. aureus only observed in fish fed on 1 and 2% FLP-supplemented diets. The weight gain values in the 1 and 2% FLP groups were higher than the other treatments (P < 0.05). Feed conversion ratio (FCR) improved with increasing FLP level in diet in a dose-response manner (P < 0.05). By considering serum and mucosal bactericidal activities against different pathogenic bacteria, the supplementation of 2% FLP in diet is recommended for C. carpio during grow-out phase.
Asunto(s)
Adiantum , Bacterias/crecimiento & desarrollo , Carpas/inmunología , Moco/inmunología , Muramidasa/inmunología , Preparaciones de Plantas/farmacología , Suero , Piel/inmunología , Animales , Carpas/sangre , Carpas/crecimiento & desarrollo , Catalasa/inmunología , Dieta/veterinaria , Inmunoglobulinas/sangre , Moco/enzimología , Muramidasa/sangre , Hojas de la Planta , Polvos , Superóxido Dismutasa/inmunologíaRESUMEN
In the current study, we investigated the effect of a probiotic bacterium (Lactobacillus rhamnosus ATCC 7469) microencapsulated with alginate and hi-maize starch and coated with chitosan on improving growth factors, body composition, blood chemistry, and the immune response of rainbow trout (initial weight: 18.41 ± 0.32 g). Four experimental diets were formulated to feed fish for 60 days. They were control diet without any additive (C), diet added with beads without probiotic (E), a probiotic sprayed to the diet (L.r), and encapsulated probiotic supplemented diet (E-L.r). The results indicated that feeding with E-Lr significantly improved weight gain (84.98 g) and feed conversion ratio (0.95) compared to the other groups (P < 0.05). Also, fish fed E-Lr diet had a significantly higher value of whole-body protein (17.51%), total protein in the blood (4.98 g/dL), lysozyme (30.66 U/mL), alternative complement pathway hemolytic activity (134 U/mL), superoxide dismutase (203 U/mg protein), and catalase (528.33 U/mg protein) (P < 0.05) as compared to those fed the control diet. Similarly, a higher relative expression of immune-related genes such as interleukin-1 (Il-1) and tumor necrosis factor-alpha (TNF-1α) were reported in those fed E-L.r and L.r diets respectively. Interestingly, the fish fed dietary E-L.r had a significantly lower value of lipid in the whole body (4.82%) and cholesterol in the blood (160.67%) in comparison with those fed the control diet (P < 0.05). At the end of the experiment, all groups were challenged by Yersinia ruckeri where the survival rate of rainbow trout fed dietary E-L.r (70.36%) was statistically higher than that of the others (P < 0.05). Overall, the results suggested that encapsulated probiotic Lact. rhamnosus ATCC 7469 acted better than unencapsulated probiotic and has a potential to improve growth performance, flesh quality, and the immune response of rainbow trout.
Asunto(s)
Enfermedades de los Peces/terapia , Proteínas de Peces/genética , Regulación de la Expresión Génica/efectos de los fármacos , Lacticaseibacillus rhamnosus/fisiología , Oncorhynchus mykiss/inmunología , Probióticos/farmacología , Yersiniosis/terapia , Alginatos/química , Alimentación Animal/análisis , Animales , Composición Corporal/efectos de los fármacos , Catalasa/genética , Catalasa/inmunología , Encapsulación Celular/métodos , Células Inmovilizadas , Quitosano/química , Colesterol/sangre , Vía Alternativa del Complemento/efectos de los fármacos , Dieta , Resistencia a la Enfermedad/efectos de los fármacos , Resistencia a la Enfermedad/genética , Resistencia a la Enfermedad/inmunología , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/microbiología , Proteínas de Peces/inmunología , Regulación de la Expresión Génica/inmunología , Interleucina-1/genética , Interleucina-1/inmunología , Muramidasa/genética , Muramidasa/inmunología , Oncorhynchus mykiss/crecimiento & desarrollo , Oncorhynchus mykiss/microbiología , Superóxido Dismutasa/genética , Superóxido Dismutasa/inmunología , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunología , Aumento de Peso/efectos de los fármacos , Yersiniosis/inmunología , Yersiniosis/microbiología , Yersinia ruckeri/efectos de los fármacos , Yersinia ruckeri/crecimiento & desarrollo , Yersinia ruckeri/patogenicidadRESUMEN
In this study, the effects of dietary myo-inositol on the skin mucosal immunity and growth of taimen (Hucho taimen) fry were determined. Triplicate groups of 500 fish (initial weight 5.58 ± 0.15 g) were fed different diets containing graded levels of myo-inositol (28.75, 127.83, 343.83, 565.81, and 738.15 mg kg-1) until satiation for 56 days. Thereafter, the nonspecific skin mucus immune parameters, antioxidative capacity, and growth performance were measured. The skin mucus protein and the activities of alkaline phosphatase were significantly higher than those in the control group (P < 0.05). However, there were no significant differences in lysozyme activity among the treatments (P > 0.05). The antimicrobial activity and minimum inhibitory concentration of the skin mucus were increased significantly by myo-inositol supplementation (P < 0.05). The superoxide dismutase, catalase, and glutathione peroxidase activities were significantly elevated in the treatment groups (P < 0.05), whereas the malondialdehyde contents were significantly decreased (P < 0.05). Low-level myo-inositol (28.75 mg kg-1) led to a significantly lower weight gain, feed efficiency, condition factor, and survival rate compared with the other treatments (P < 0.05). In conclusion, dietary myo-inositol deficiency (28.75 mg kg-1) adversely affects the skin mucus immune parameters, antioxidative capacity, and growth performance of Hucho taimen fry.
Asunto(s)
Carpas/inmunología , Suplementos Dietéticos , Inmunidad Mucosa/efectos de los fármacos , Inositol/farmacología , Moco/efectos de los fármacos , Piel/efectos de los fármacos , Complejo Vitamínico B/farmacología , Aeromonas hydrophila/crecimiento & desarrollo , Alimentación Animal , Animales , Carpas/genética , Carpas/crecimiento & desarrollo , Carpas/metabolismo , Catalasa/inmunología , Dieta/veterinaria , Glutatión Peroxidasa/inmunología , Moco/enzimología , Moco/inmunología , Piel/enzimología , Piel/inmunología , Superóxido Dismutasa/inmunología , Yersinia ruckeri/crecimiento & desarrolloRESUMEN
l-theanine is a water-soluble non-proteinous amino acid mainly found in green tea leaves. Despite the availability of abundant literature on green tea, studies on the use of l-theanine as a feed additive in animals, and especially broilers are limited. The objective of this study was, therefore, to evaluate the effect of different dietary levels of l-theanine on meat quality, growth performance, immune response, and blood metabolites in broilers. A total of 400 day-old broiler chicks were randomly divided into four treatment groups using a completely randomized design; C-control, basal diet; 100LT-basal diet + 100 mg l-theanine/kg diet; 200LT-basal diet + 200 mg l-theanine/kg diet; and 300LT-basal diet + 300 mg l-theanine/kg diet. Results revealed that the intermediate level of l-theanine (200 mg/kg diet) showed better results in terms of body weight gain (BWG), feed consumed (FC), and feed conversion ratio (FCR) as compared with the other supplemented groups and the control. The live weight eviscerated weight and gizzard weight were higher in all l-theanine levels as compared to those of the control group. Increased weight (p ≤ 0.05) of spleen and bursa were found in group 200LT (200 mg l-theanine/kg diet). Concerning meat color parameters, values for yellowness (b*), and redness (a*) were greater in l-theanine-supplemented groups than the control. Supplementing broiler diet with l-theanine reduced (p = 0.02) total serum cholesterol contents while increased HDL. Further analysis revealed lower relative serum cytokines (IL-2 and INF-γ) and reduced mRNA expression of TNF-α and IL-6 in thymus, and IFN-γ and IL-2 in spleen in the treated group. Moreover, supplementation with 200 mg/kg of l-theanine improved antioxidant status in blood by increasing SOD, GSH-Px, and relative CAT levels. It is concluded that the optimum supplementation level of l-theanine is 200 mg/kg of diet because it resulted in improved performance parameters in broilers. However, higher levels of l-theanine (300 mg/kg diet) may have deleterious effects on performance and health of broiler chickens.
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Pollos/inmunología , Suplementos Dietéticos , Glutamatos/administración & dosificación , Carne/análisis , Músculo Esquelético/efectos de los fármacos , Aumento de Peso/efectos de los fármacos , Alimentación Animal/análisis , Animales , Bolsa de Fabricio/efectos de los fármacos , Bolsa de Fabricio/inmunología , Bolsa de Fabricio/metabolismo , Catalasa/genética , Catalasa/inmunología , Pollos/genética , Pollos/crecimiento & desarrollo , LDL-Colesterol/sangre , Calidad de los Alimentos , Regulación de la Expresión Génica/inmunología , Glutatión Peroxidasa/genética , Glutatión Peroxidasa/inmunología , Inmunidad Innata/efectos de los fármacos , Interferón gamma/genética , Interferón gamma/inmunología , Interleucina-2/genética , Interleucina-2/inmunología , Interleucina-6/genética , Interleucina-6/inmunología , Músculo Esquelético/inmunología , Músculo Esquelético/metabolismo , Tamaño de los Órganos/efectos de los fármacos , Bazo/efectos de los fármacos , Bazo/inmunología , Bazo/metabolismo , Superóxido Dismutasa/genética , Superóxido Dismutasa/inmunología , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
The present study was carried out to investigate the anti-arthritic activity of Berberis aristata hydroalcoholic extract (BAHE) in formaldehyde-induced arthritis and adjuvant-induced arthritis (AIA) model. Arthritis was induced by administration of either formaldehyde (2% v/v) or CFA into the subplantar surface of the hind paw of the animal. In formaldehyde-induced arthritis and AIA, treatment of BAHE at doses 50, 100 and 200 mg/kg orally significantly decreased joint inflammation as evidenced by decrease in joint diameter and reduced inflammatory cell infiltration in histopathological examination. BAHE treatment demonstrated dose-dependent improvement in the redox status of synovium (decrease in GSH, MDA, and NO levels and increase in SOD and CAT activities). The beneficial effect of BAHE was substantiated with decreased expression of inflammatory markers such as IL-1ß, IL-6, TNF-R1, and VEGF by immunohistochemistry analysis in AIA model. BAHE increased HO-1/Nrf-2 and suppressed NF-κB mRNA and protein expression in adjuvant immunized joint. Additionally, BAHE abrogated degrading enzymes, as there was decreased protein expression of MMP-3 and -9 in AIA. In conclusion, we demonstrated the anti-arthritic activity of Berberis aristata hydroalcoholic extract via the mechanism of inhibition of NF-κB and activation of Nrf-2/HO-1.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Artritis Experimental/tratamiento farmacológico , Berberis/química , Hemo Oxigenasa (Desciclizante)/inmunología , Factor 2 Relacionado con NF-E2/inmunología , FN-kappa B/inmunología , Extractos Vegetales/farmacología , Administración Oral , Animales , Artritis Experimental/inducido químicamente , Artritis Experimental/inmunología , Artritis Experimental/patología , Catalasa/genética , Catalasa/inmunología , Relación Dosis-Respuesta a Droga , Formaldehído , Adyuvante de Freund , Regulación de la Expresión Génica , Glutatión/agonistas , Glutatión/inmunología , Goma Arábiga , Hemo Oxigenasa (Desciclizante)/genética , Masculino , Malondialdehído/antagonistas & inhibidores , Malondialdehído/inmunología , Factor 2 Relacionado con NF-E2/genética , FN-kappa B/genética , Óxido Nítrico/antagonistas & inhibidores , Óxido Nítrico/inmunología , Ratas , Ratas Wistar , Transducción de Señal , Superóxido Dismutasa/genética , Superóxido Dismutasa/inmunología , Membrana Sinovial/efectos de los fármacos , Membrana Sinovial/inmunología , Membrana Sinovial/patología , Tarso Animal/efectos de los fármacos , Tarso Animal/inmunología , Tarso Animal/patologíaRESUMEN
OBJECTIVE: To investigate the changes of B and T lymphocyte attenuator (BTLA), reactive oxygen species (ROS), reactive nitrogen species (RNS), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), total antioxidative capacity (TAOC) in the patients with ankylosing spondylitis (AS) and the effect of Xinfeng capsule (XFC) on them. METHODS: AS patients (n=140) were randomly divided into two groups, XFC group (3 tablets each time, tid, n=70) and salicylazosulfapyridine (SASP) group (4 pills each time, bid, n=70). Continuous treatment lasts 3 months. The study also enrolled 60 healthy volunteers as a control group. Flow cytometry was used to test BTLA expression. ELISA was performed to detect the oxidative stress indicators (ROS, RNS, MDA, SOD, CAT, TAOC) and cytokines (IL-4, IL-10, IL-1ß, TNF-α). Western blotting was adopted to examine the blood sedimentation (ESR). HITACHI 7060 automatic biochemical analyzer was used to determine the level of high sensitive C-reactive protein (Hs-CRP). RESULTS: Clinical efficacy of XFC group was significantly better than that of SASP group (P<0.01). Compared with the healthy control group, AS patients had significantly lower BTLA expression in CD3(+) T cells and CD4(+) T cells from the peripheral blood (P<0.01 or P<0.05), the decreased levels of SOD, CAT and TAOC, and significantly increased ROS, RNS and MDA values (P<0.01 or P<0.05). In addition, the levels of serum IL-1ß, TNF-α, ESR and Hs-CRP were significantly higher (P<0.01) and IL-4, IL-10 were significantly lower in AS patients (P<0.01 or P<0.05). Compared with pre-treatment, both XFC and SASP significantly elevated the expressions of BTLA(+)CD3(+) T, BTLA(+)CD4(+) T, BTLA, SOD, TAOC, IL-4, SF-36 (PF, SF, RP, RE, BP, MH, VT, GH) eight dimension scores, and reduced ROS, MDA, TNF-α, ESR, Hs-CRP, VAS, BASDAI, BASFI and BAS-G in the peripheral blood (P<0.01 or P<0.05). The differences between XFC group and SASP group were statistically significant (P<0.01 or P<0.05). Pearson correlation analysis showed that BTLA expression level in the peripheral blood was positively correlated with SOD, RP, BP, SF and RE. BTLA(+)CD3(+) T cells and BTLA*CD4(+) T cells were significantly negatively correlated with ROS, MDA, IL-1ß, TNF-α, ESR, VAS and BASDAI, and they were positively correlated with TAOC, IL-4 and IL-10. BTLA(+)CD3(+) T cells were significantly negatively correlated with RNS, Hs-CRP and BASFI; BTLA(+)CD4(+) T cells were positively correlated with CAT. CONCLUSION: XFC can improve BTLA expression in the peripheral blood of AS patients and regulate negatively the activation and proliferation of T cells.
Asunto(s)
Medicamentos Herbarios Chinos/uso terapéutico , Estrés Oxidativo/efectos de los fármacos , Espondilitis Anquilosante/tratamiento farmacológico , Linfocitos T/efectos de los fármacos , Adolescente , Adulto , Antiinflamatorios no Esteroideos/uso terapéutico , Cápsulas , Catalasa/inmunología , Catalasa/metabolismo , Citocinas/inmunología , Citocinas/metabolismo , Citometría de Flujo , Humanos , Recuento de Linfocitos , Masculino , Malondialdehído/inmunología , Malondialdehído/metabolismo , Persona de Mediana Edad , Estrés Oxidativo/inmunología , Fitoterapia/métodos , Especies de Nitrógeno Reactivo/inmunología , Especies de Nitrógeno Reactivo/metabolismo , Especies Reactivas de Oxígeno/inmunología , Especies Reactivas de Oxígeno/metabolismo , Receptores Inmunológicos/inmunología , Receptores Inmunológicos/metabolismo , Espondilitis Anquilosante/inmunología , Sulfasalazina/uso terapéutico , Superóxido Dismutasa/inmunología , Superóxido Dismutasa/metabolismo , Linfocitos T/inmunología , Linfocitos T/metabolismo , Factores de Tiempo , Resultado del Tratamiento , Adulto JovenRESUMEN
We evaluated the protective efficacy of the polyphenolic fraction from virgin coconut oil (PV) against adjuvant induced arthritic rats. Arthritis was induced by intradermal injection of complete Freund's adjuvant. The activities of inflammatory, antioxidant enzymes and lipid peroxidation were estimated. PV showed high percentage of edema inhibition at a dose of 80mg/kg on 21st day of adjuvant arthritis and is non toxic. The expression of inflammatory genes such as COX-2, iNOS, TNF-α and IL-6 and the concentration of thiobarbituric acid reactive substance were decreased by treatment with PV. Antioxidant enzymes were increased and on treatment with PV. The increased level of total WBC count and C-reactive protein in the arthritic animals was reduced in PV treated rats. Synovial cytology showed that inflammatory cells and reactive mesothelial cells were suppressed by PV. Histopathology of paw tissue showed less edema formation and cellular infiltration on supplementation with PV. Thus the results demonstrated the potential beneficiary effect of PV on adjuvant induced arthritis in rats and the mechanism behind this action is due to its antioxidant and anti-inflammatory effects.
Asunto(s)
Antiinflamatorios/uso terapéutico , Antioxidantes/uso terapéutico , Artritis Experimental/tratamiento farmacológico , Aceites de Plantas , Polifenoles/uso terapéutico , Animales , Antiinflamatorios/farmacología , Antioxidantes/farmacología , Artritis Experimental/inmunología , Artritis Experimental/patología , Proteína C-Reactiva/inmunología , Catalasa/inmunología , Aceite de Coco , Ciclooxigenasa 2/genética , Pie/patología , Glutatión Peroxidasa/inmunología , Interleucina-6/inmunología , Recuento de Leucocitos , Masculino , Óxido Nítrico/sangre , Óxido Nítrico Sintasa de Tipo II/genética , Polifenoles/farmacología , Prostaglandina-Endoperóxido Sintasas/inmunología , ARN Mensajero/metabolismo , Ratas Wistar , Superóxido Dismutasa/inmunología , Membrana Sinovial/patología , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
In view of well-established immunomodulatory properties of Lactobacillus, present investigation was carried out to evaluate antioxidant and anti-inflammatory potential of Lactobacillus casei and Lactobacillus acidophilus, against inflammatory pathway and oxidative stress developed in an experimental model of arthritis. Collagen-induced arthritis (CIA) model was used. Oral administration of L. casei, L. acidophilus, standard antiarthritic drug indomethacin, and vehicle were started after induced arthritis and continued up to day 28. Interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-1ß, IL-17, IL-4, and IL-10 levels were estimated in serum. In parallel, oxidative stress parameters were also measured from synovial effsuate. All rats were graded for arthritis score at the end of each week. L. casei, L. acidophilus, and indomethacin treatment significantly downregulated proinflammatory and upregulated anti-inflammatory cytokines at P<0.0001. They have significantly decreased oxidative stress in synovial effsuate (P<0.0001) and also arthritis score (P<0.05). Protection provided by L. casei and L. acidophilus was more pronounced than that of indomethacin. These lines of evidence suggest that L. casei and L. acidophilus exert potent protective effect against CIA. It further establishes effective anti-inflammatory and antioxidant properties of Lactobacillus. However, additional clinical investigations are needed to prove the efficacy of Lactobacillus in treatment/management of rheumatoid arthritis.
Asunto(s)
Artritis Experimental/inmunología , Inflamación/inmunología , Lacticaseibacillus casei/inmunología , Lactobacillus acidophilus/inmunología , Transducción de Señal/inmunología , Animales , Antioxidantes/metabolismo , Artritis Experimental/microbiología , Artritis Experimental/prevención & control , Catalasa/inmunología , Catalasa/metabolismo , Glutatión/inmunología , Glutatión/metabolismo , Glutatión Peroxidasa/inmunología , Glutatión Peroxidasa/metabolismo , Interacciones Huésped-Patógeno/inmunología , Inflamación/microbiología , Interleucina-10/sangre , Interleucina-10/inmunología , Interleucina-17/sangre , Interleucina-17/inmunología , Interleucina-1beta/sangre , Interleucina-1beta/inmunología , Interleucina-4/sangre , Interleucina-4/inmunología , Interleucina-6/sangre , Interleucina-6/inmunología , Lactobacillus acidophilus/fisiología , Lacticaseibacillus casei/fisiología , Peroxidación de Lípido/efectos de los fármacos , Peroxidación de Lípido/inmunología , Masculino , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/inmunología , Probióticos/administración & dosificación , Probióticos/farmacología , Ratas , Ratas Wistar , Transducción de Señal/efectos de los fármacos , Superóxido Dismutasa/inmunología , Superóxido Dismutasa/metabolismo , Factores de Tiempo , Factor de Necrosis Tumoral alfa/sangre , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
CONTEXT: Leptadenia reticulata Linn. (Asclpiadaceae) commonly known as "dodi," is an Indian medicinal plant which is known to have ethno-medical uses such as stimulant, tonic, immunostimulant and is one of the ingredient in ayurvedic formulation called as "Chawanprash," which is widely used in India to increase the strength of immune system. OBJECTIVE: The aim of present study is to evaluate immunomodulatory and antioxidant activity of ethanolic extract of L. reticulata L. leaves in rodents. METHODS: Haemagglutinating antibody (HA) titre, haematological profile (Hb, WBC, RBC), lipid peroxidation (LPO), reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), delayed type of hypersensitivity (DTH) response, neutrophil adhesion test and carbon clearance assay were determined by in vivo experiments. RESULTS: The evaluation of immunomodulatory potential of L. reticulata (100, 200 mg/kg, p.o.) evoked a significant dose-dependent increase in antibody titre values; DTH reaction induced by SRBC and potentiated percentage neutrophil adhesion to nylon fibers as well as phagocytosis in carbon clearance assay. Also it caused significant increase in haematological profile, GSH, SOD, CAT activity and significantly decreased LPO levels in cyclophosphamide-induced immunosuppressed rats. CONCLUSION: The results obtained in this study indicate that L. reticulata possesses potential immunomodulatory and antioxidant activity and can play a major role in reducing the risk to develop immunodeficiency disorders.
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Antioxidantes/farmacología , Apocynaceae/química , Inmunidad Celular/efectos de los fármacos , Inmunidad Humoral/efectos de los fármacos , Factores Inmunológicos/farmacología , Extractos Vegetales/farmacología , Hojas de la Planta/química , Animales , Antioxidantes/química , Catalasa/sangre , Catalasa/inmunología , Adhesión Celular/efectos de los fármacos , Adhesión Celular/inmunología , Femenino , Glutatión/sangre , Glutatión/inmunología , Hipersensibilidad Tardía/sangre , Hipersensibilidad Tardía/tratamiento farmacológico , Hipersensibilidad Tardía/inmunología , Factores Inmunológicos/química , Peroxidación de Lípido/efectos de los fármacos , Peroxidación de Lípido/inmunología , Masculino , Ratones , Neutrófilos/inmunología , Neutrófilos/metabolismo , Extractos Vegetales/química , Ratas , Ratas Wistar , Superóxido Dismutasa/sangre , Superóxido Dismutasa/inmunologíaRESUMEN
Catalase is one of the key antioxidant enzymes and it appears to be involved in protection against immune infection and oxidative stress. Here, two catalase cDNAs (ChCat-1 and ChCat-2) were isolated from hemocytes of Crassostrea hongkongensis using SSH and RACE. The full-length cDNAs of ChCat-1 and ChCat-2 are 1913 and 2466 bp in length, encoding proteins of 515 and 511 amino acids, respectively. Multiple alignments of amino acid sequences revealed that both ChCat-1 and ChCat-2 possess several characteristic features of the catalase family of enzymes, including one proximal active site signature, one heme-ligand signature, and three catalytic amino acid residues (His(72), Asn(145) and Tyr(355)). Phylogenetic analysis indicates that these two catalases may share a common ancestral gene and result from a gene duplication event following the divergence of bivalves and gastropods. Constitutive expression of ChCat-1 and ChCat-2 was observed in all tissues studied, with highest levels of expression in gill and muscle, respectively. The expression of both genes was inducible by bacterial infection, and reached the maximum at 8 h (9.0-fold) and 12 h (2.3-fold) post-infection, respectively. Furthermore, both the purified ChCat-1 and ChCat-2 protein displayed a strong catalase activity, and S2 cells carrying ChCat-1 or ChCat-2 showed a higher degree of resistance to H(2)O(2) than that of control cells. In a word, this is the first report of the presence of two catalase genes in a single marine bivalve, and our results highlight the involvement of both ChCat-1 and ChCat-2 in host protection against pathogen infection and oxidative stress in C. hongkongensis.
Asunto(s)
Catalasa/genética , Catalasa/inmunología , Crassostrea/inmunología , Regulación de la Expresión Génica/inmunología , Estrés Oxidativo/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Western Blotting , Catalasa/metabolismo , Crassostrea/enzimología , Crassostrea/microbiología , Cartilla de ADN/genética , ADN Complementario/genética , Genes Duplicados/genética , Branquias/metabolismo , Hemocitos/metabolismo , Peróxido de Hidrógeno , Datos de Secuencia Molecular , Músculos/metabolismo , Filogenia , Análisis de Secuencia de ADN , Homología de SecuenciaRESUMEN
Recent studies revealed that antioxidant enzymes play important roles in antioxidant responses caused by metabolic process or pathogen invasion. Catalase is one of these key enzymes which has been characterized and highly conserved from invertebrates to vertebrates. In the present study, a full-length cDNA sequence of catalase was cloned from the hemocyte suppression subtractive hybridization library of the crab Scylla paramamosain. The Sp-catalase (Sp-CAT) cDNA sequence contained 2551bp with an open reading frame of 1551bp encoding 517 amino acid residues. The conserved catalytic active residues His-71, Asn-144 and Tyr-354 were predicted in the amino acid sequence of Sp-CAT. The deduced Sp-CAT protein had a calculated molecular mass of 59 kDa with an estimated isoelectric point of 6.4. Multiple alignment analysis revealed that the deduced amino acid sequence of Sp-CAT shared high identity (75.4%) with those of other species. The Sp-CAT mRNA transcripts were demonstrated in multiple tissues of normal S. paramamosain. After LPS challenge, the expression level of Sp-CAT gene was increased significantly in hemocyte at 3 and 6 h, and in hepatopancreas at 6 h, respectively, determined by quantitative real-time PCR. Furthermore, the activities of CAT and SOD were also measured in different tissues and serum after LPS challenge. The CAT activity was significantly increased at 3, 6, 24 and 48 h in hemocyte lysate, at 3 h in serum, and at 24 and 48 h in hepatopancreas after LPS challenge. In addition, the SOD activity was significantly induced at 3 and 6 h in hemocyte lysate, 3 and 12 h in serum, 12 and 48 h in hepatopancreas post LPS stimulation, indicating a tissue and time-dependent antioxidant response in the crab. Taken together, these data demonstrated that a strong antioxidant response occurred in the LPS-challenged crab, which might be involved in the protection of host against microbial infections.
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Adyuvantes Inmunológicos/farmacología , Braquiuros , Catalasa , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Lipopolisacáridos/farmacología , Superóxido Dismutasa/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Braquiuros/efectos de los fármacos , Braquiuros/enzimología , Braquiuros/genética , Catalasa/química , Catalasa/genética , Catalasa/inmunología , Clonación Molecular , Perfilación de la Expresión Génica , Hemocitos/efectos de los fármacos , Hemocitos/inmunología , Hepatopáncreas/efectos de los fármacos , Hepatopáncreas/inmunología , Modelos Moleculares , Datos de Secuencia Molecular , Estructura Terciaria de ProteínaRESUMEN
Deep pulmonary delivery of an influenza ISCOMATRIX vaccine has previously been shown to induce a combined mucosal and systemic antibody response. To explore whether this combined response is influenced by intrinsic properties of the component antigen, we examined the efficacy of deep pulmonary delivery of ISCOMATRIX vaccines containing different recombinant antigens, specifically gB glycoprotein from cytomegalovirus and a fragment of catalase from Helicobacter pylori. Both these vaccines induced antigen-specific mucosal and systemic immunity, as well as antigen-specific proliferative cellular responses. Pulmonary immunisation with ISCOMATRIX vaccines may therefore be a generic way of inducing combined systemic and mucosal immunity.
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Adyuvantes Inmunológicos/administración & dosificación , Antígenos Bacterianos/inmunología , Antígenos Virales/inmunología , Colesterol/administración & dosificación , Fosfolípidos/administración & dosificación , Saponinas/administración & dosificación , Administración por Inhalación , Animales , Anticuerpos Antibacterianos/sangre , Anticuerpos Antivirales/sangre , Catalasa/inmunología , Proliferación Celular , Combinación de Medicamentos , Femenino , Helicobacter pylori/enzimología , Inyecciones Subcutáneas , Linfocitos/inmunología , Ovinos , Vacunas Sintéticas/inmunología , Proteínas del Envoltorio Viral/inmunologíaRESUMEN
OBJECTIVE: To investigate the effects of the two-valence vaccine consisting of Helicobacter pylori (H. pylori) catalase and urease subunit UreB in preventing H. pyloriinfection in mice. METHODS: C57BL/6 mice were divided into 7 groups and immunized with intragastric administration of catalase and UreB (both 100 microg) plus cholera toxin (CT, 2 microg), catalase (100 microg) plus CT (2 microg), UreB (100 microg) plus CT (2 microg), catalase (100 microg), UreB (100 microg), CT (2 microg), or PBS, respectively, once a week for 4 consecutive weeks. Two weeks after the last immunization, all the mice were challenged by live H. pylori, and sacrificed 4 weeks after the challenge to obtain the gastric mucosa samples for detecting H. pylori using semi-quantitative bacterial culture assay. RESULTS: The total protection rate in mice immunized with the two-valence vaccine, single-valence vaccine of catalase, and single-valence vaccine of UreB was 83.3% (20/24), 41.7% (10/24) and 54.2% (13/24), respectively, and the rate in the other 4 groups were all 0. The H. pyloricolony density in mice with vaccination was significantly lower than that of other 4 groups (P<0.05). The total protection rate and H. pylori colony density differed significantly between the two-valence vaccination group and the single-valence vaccination groups (P<0.05). CONCLUSION: The two-valence vaccine consisting of catalase, UreB and adjuvant has better immunoprotective effects than the single-valence vaccines.
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Vacunas Bacterianas/inmunología , Catalasa/inmunología , Infecciones por Helicobacter/prevención & control , Helicobacter pylori/inmunología , Ureasa/inmunología , Animales , Proteínas Bacterianas/inmunología , Infecciones por Helicobacter/inmunología , Helicobacter pylori/enzimología , Inmunización/métodos , Masculino , Ratones , Ratones Endogámicos C57BL , Distribución AleatoriaRESUMEN
We have developed a two-step selection protocol to generate a population of Chinese hamster ovary (CHO) cell variants that are plasmalogen-deficient, but contain intact, functional peroxisomes (plasmalogen-/peroxisome+). This involved sequential exposures of a mutagenized cell population to photodynamic damage by using two different pyrene-labelled sensors, 9-(1'-pyrene)nonanol and 12-(1'-pyrene)dodecanoic acid. By this procedure we generated several isolates, all except one of which displayed a severe decrease in plasmalogen biosynthesis. Further characterization of one of the plasmalogen-deficient isolates, NRel-4, showed that it contained intact, functional peroxisomes. Whole-cell homogenates from NRel-4 displayed severely decreased dihydroxyacetone phosphate acyltransferase, which catalyses the first step in plasmalogen biosynthesis. NRel-4 and another, recently described, plasmalogen-deficient cell line, NZel-1 [Nagan, Hajra, Das, Moser, Moser, Lazarow, Purdue and Zoeller (1997) Proc. Natl. Acad. Sci. U.S. A. 94, 4475-4480] were hypersensitive to singlet oxygen, supporting the notion of plasmalogens as radical oxygen scavengers. Wild-type-like resistance could be conferred on NRel-4 upon restoration of plasmalogen content by supplementation with a bypass compound, sn-1-hexadecylglycerol. NRel-4 and other plasmalogen-/peroxisome+ strains will allow us to examine further the role of ether lipids in cellular functions without complications associated with peroxisome deficiency, and might serve as an animal cell model for certain forms of the human genetic disorder rhizomelic chondrodysplasia punctata.
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Aciltransferasas/deficiencia , Microcuerpos/enzimología , Plasmalógenos/biosíntesis , Animales , Células CHO , Catalasa/inmunología , Supervivencia Celular/efectos de los fármacos , Condrodisplasia Punctata Rizomélica/genética , Cricetinae , Ácidos Grasos/farmacología , Inmunohistoquímica , Microcuerpos/patología , Mutagénesis/genética , Fosfolípidos/análisis , Pirenos/farmacología , Rayos Ultravioleta/efectos adversosRESUMEN
Ten complementation groups of generalized peroxisome biogenesis disorders (PBD), (excluding rhizomelic chondrodysplasia punctata) have been identified using complementation analysis. Four of the genes involved have been identified using two different methods of (1) genetic functional complementation of peroxisome deficient CHO cell mutants and (2) homology searches for human dbEST, based on yeast genes involved in peroxisome biogenesis (PEX genes). We report here the first identification of a new complementation group which is genetically different from peroxisome deficient CHO mutants. There were no complementations by the human PEX 13 gene. The nature of the related gene is being investigated.
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Prueba de Complementación Genética , Proteínas de la Membrana/genética , Trastorno Peroxisomal/genética , Animales , Células CHO , Catalasa/inmunología , Mapeo Cromosómico , Clonación Molecular , Cricetinae , ADN Complementario/genética , Fibroblastos , Técnica del Anticuerpo Fluorescente , Humanos , Lactante , Masculino , Fenotipo , Transfección/genéticaRESUMEN
The efficacy of an orogastric vaccine comprised of purified Helicobacter pylori catalase plus the mucosal adjuvant cholera toxin (CT) was examined with both the Helicobacter felis and H. pylori mouse models with BALB/c mice. Native H. pylori catalase (200 microg) plus CT was initially used as a vaccine antigen in the H. felis mouse model and protected 80% (8 of 10) of the challenged animals, while all control animals were infected (20 of 20). In a follow-up experiment, recombinant H. pylori catalase plus CT was used for immunization, and groups of mice were challenged with the Sydney strain of H. pylori. Immunization with recombinant catalase protected a significant proportion (9 of 10) of the mice from H. pylori challenge, indicating that this enzyme should be considered as a candidate for a future vaccine. This study provides the first available data on the efficacy of protective immunization with the new Sydney strain of H. pylori in a mouse model. These data also provide indirect evidence that proteins which are normally intracellular, such as catalase, may be present on the surface of H. pylori and thus may provide targets for immunization.