RESUMO
We evaluated changes in BAX and BCL2 expression levels after spinal cord ischemia/reperfusion injury (SCII) and hypothermia during operations in rats. Eighty rats were divided into four groups: Group A (N = 20, 18°C); Group B (N = 20, 28°C); Group C (N = 20, room temperature); and Group D (N = 20, sham operation control). Spinal cord ischemia was induced for 90 min. Hypothermia was induced 15 min before, and maintained during ischemia, followed by heating to normothermia for 30 min after reperfusion. Motor function of the lower limbs was evaluated according to the Tarlov score at 72 and 168 h. For each rat, spinal cord samples were taken at 6, 24, 72 h, and 1 week to evaluate the histopathological changes, neuronal apoptosis, and BAX and BCL2 expression levels. Compared with normothermia, hypothermia significantly improved hind limb function; Group B achieved a higher score than Group A. Group D showed no neurologic deficiency, while the other groups showed various degrees. Group C exhibited greater neuronal apoptosis, higher BAX expression, but lower BCL2 expression than the other groups. Compared with Group A, BAX was expressed less and BCL2 more in Group B, and there was less apoptosis in Group B. Hypothermia preserves hind limb motor function and reduces neuronal death, thereby protecting rats from SCII. The spinal cord may be protected from SCII by inhibition of BAX and activation of BCL2. However, deep hypothermia may inhibit the expression of BCL2, resulting in a worse outcome than mild hypothermia.
Assuntos
Proteínas Proto-Oncogênicas c-bcl-2/genética , Traumatismo por Reperfusão/genética , Traumatismo por Reperfusão/prevenção & controle , Isquemia do Cordão Espinal/genética , Medula Espinal/metabolismo , Proteína X Associada a bcl-2/genética , Animais , Apoptose/genética , Temperatura Baixa , Regulação da Expressão Gênica , Membro Posterior/irrigação sanguínea , Membro Posterior/fisiopatologia , Masculino , Atividade Motora/fisiologia , Neurônios/metabolismo , Neurônios/patologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/fisiopatologia , Medula Espinal/patologia , Isquemia do Cordão Espinal/metabolismo , Isquemia do Cordão Espinal/patologia , Isquemia do Cordão Espinal/fisiopatologia , Proteína X Associada a bcl-2/metabolismoRESUMO
We examined the effects of co-culturing CD4+ CD25+ Treg cells with sirolimus or cyclosporin A on Treg cell proliferation and differentiation and on transforming growth factor-ß (TGF-ß) and Foxp3 expression. CD4+ CD25+ Treg cells were harvested from mononuclear cells of spleens of C57BL/6 mice using immunomagnetic beads and divided into control, sirolimus, and cyclosporine groups. Following a 96-h co-culture, Treg cells were assayed by flow cytometry. FoxP3 and TGF-ß mRNA levels and secretion were assayed by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay. Smad protein of the TGF-ß signaling pathway was assayed by western blot and its effect on CD4+ CD25+ FoxP3+ Treg cell proliferation was determined. Sirolimus-promoted differentiation and proliferation was examined using a TGF-ß neutralizing antibody. Sirolimus-treated CD4+ T cell TGF-ß secretion increased 2.5X over control levels (P < 0.01), but that of the cyclosporine group decreased marginally (P > 0.05). The CD4+ cell proportion decreased significantly (41.25 vs 69.22%, P < 0.01) and slightly (65.21 vs 69.22, P > 0.05) in the cyclosporine and sirolimus groups, respectively. T cell Foxp3 mRNA expression was significantly higher in the sirolimus-treated than in the cyclosporine (53.7 vs 40.2%, P < 0.05) and control groups (P < 0.01), but was significantly lower in the cyclosporine group than in controls (23.6 vs 40.2%, P < 0.01). Overall, sirolimus promoted CD4+ CD25+ Treg cell proliferation and growth in vitro, whereas cyclosporin A inhibited proliferation. Sirolimus might promote CD4+ CD25+ FoxP3+ regulatory T cell proliferation by inducing TGF-ß secretion in vivo.
Assuntos
Imunossupressores/farmacologia , Sirolimo/farmacologia , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T Reguladores/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Animais , Biomarcadores , Proliferação de Células/efeitos dos fármacos , Ciclosporina/farmacologia , Fatores de Transcrição Forkhead/metabolismo , Masculino , Camundongos , Proteína Smad2/metabolismo , Proteína Smad3/metabolismo , Subpopulações de Linfócitos T/efeitos dos fármacos , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Linfócitos T Reguladores/imunologiaRESUMO
The main objective of the current study was to explore the different activation mechanisms of capacitation and freeze-thawed spermatozoa. Using SDS-PAGE and Western blotting, the conversion process of boar proacrosin during freeze-thawing and capacitation of spermatozoa was analyzed. The results revealed that capacitated spermatozoa exhibited a greater fluorescence area than that of the freeze-thawed spermatozoa, which were smaller than those of the fresh group. Fresh spermatozoa displayed 45- and 35- kDa protein bands, while those of freeze-thawed andcapacitated spermatozoa displayed 45-, 35- and 28-kDa bands. In summary, these data indicate that proacrosin is activated, thus becoming α- and ß-acrosins and a 28-kDa protein during capacitation and freeze-thawing.
Assuntos
Acrosina/metabolismo , Criopreservação , Precursores Enzimáticos/metabolismo , Capacitação Espermática , Espermatozoides/metabolismo , Suínos/metabolismo , Animais , Western Blotting , Fluorescência , Masculino , Coloração e RotulagemRESUMO
To explore the relationship between Myf5 gene polymorphisms and production performance traits in Songliao white geese, we used the chicken Myf5 sequence to design primers and amplified part of the exon 1 sequence of the Songliao white goose Myf5 gene. Results of single-strand conformation polymorphism polymerase chain reaction analysis revealed polymorphisms of the amplified fragment, including three genotypes (AA, AB, and BB). Three varieties were dominated by allele A and were mainly expressed in AA genotypes. We also identified that the Myf5 gene has one single nucleotide change (AâG) on exon 1 at locus 1344, and another (GâC) at locus 1410. Analysis of variance showed significant differences between genotypes before slaughter in live weight, carcass weight, eviscerated weight, leg muscle weight, weight of the wings, and slaughter rate. There were no significant differences with respect to other growth and carcass traits evaluated.