RESUMEN
The aim of the present study was to determine the preventive effects of the polysaccharide of Larimichthys crocea swim bladder (PLCSB) on CCl4-induced hepatic damage in ICR mice. The in vitro preventive effects of PLCSB on CCl4-induced liver cytotoxic effect were evaluated in BRL 3A rat liver cells using the MTT assay. The serum levels of AST, ALT, and LDH in mice were determined using commercially available kits. The levels of IL-6, IL-12, TNF-α, and IFN-γ were determined using ELISA kits. The pathological analysis of hepatic tissues was performed with H and E staining, and the gene and protein expressions were determined by RT-PCR and Western blotting, respectively. PLCSB (20 μg·mL(-1)) could increase the growth of BRL 3A rat liver cells treated with CCl4. The serum levels of AST, ALT, and LDH were significantly decreased when the mice were treated with two doses of PLCSB, compared with the control mice (P < 0.05). PLCSB-treated groups also showed reduced levels of the serum pro-inflammatory cytokines IL-6, IL-12, TNF-α, and IFN-γ. PLCSB could decrease the liver weight, compared to the CCl4-treated control mice. The histopathology sections of liver tissues in the 100 mg·kg(-1) PLCSB group indicated that the animals were recovered well from CCl4 damage, but the 50 mg·kg(-1) PLCSB group showed necrosis to a more serious extent. The 100 mg·kg(-1) PLCSB group showed significantly decreased mRNA and protein expression levels of NF-κB, iNOS, and COX-2, and increased expression of IκB-α compared with the CCl4-treated control group. In conclusion, PLCSB prevented from CCl4-induced hepatic damage in vivo.
Asunto(s)
Animales , Masculino , Estructuras Animales , Química , Productos Biológicos , Farmacología , Usos Terapéuticos , Tetracloruro de Carbono , Intoxicación por Tetracloruro de Carbono , Quimioterapia , Metabolismo , Patología , Enfermedad Hepática Inducida por Sustancias y Drogas , Metabolismo , Patología , Ciclooxigenasa 2 , Metabolismo , Citocinas , Sangre , Proteínas I-kappa B , Metabolismo , Mediadores de Inflamación , Sangre , Hígado , Metabolismo , Patología , Ratones Endogámicos ICR , Inhibidor NF-kappaB alfa , FN-kappa B , Metabolismo , Necrosis , Óxido Nítrico Sintasa de Tipo II , Metabolismo , Perciformes , Polisacáridos , Farmacología , Usos Terapéuticos , ARN Mensajero , MetabolismoRESUMEN
Objective To establish a spontaneous abortion rat model for a syndrome in traditional chinese medicine, kidney deficiency, and observe the changes of physiological indicators and related cytokine expression in the model. Methods 40 female and 20 male rats were used in this study. The female and male rats were mated (mating ratio 2:1). The day of vaginal smear with a large number of sperm was considered as the first day of pregnancy. The rats were randomly divided into control group and model group. The model group received 450 mg/kg hydroxyurea every day. Mifepristone was given on the eighth day in a dose of 3.75 mg/kg. The diet amount, the diameter index of kidney, ovary and embryos were analyzed. The mRNA expression of Th1/Th2 cytokine was detected by RT-PCR, and the expression of co-stimulating factors CD80, CD86, CD28, CTLA-4 were determined by flow cytometry.Results Comparing the model group with control group on the eighth day, there were significant differences between the model and control groups in quantity of food and water intake, and weight increase (P<0.05), and also in the embryonic diameter index, average of abortion rate, Th1/Th2 type cytokines, co-stimulating factor CD80, CD86, CD28, and CTLA-4 (P<0.05). Conclusion A rat model of spontaneous abortion due to kidney deficiency can be successfully established with hydroxyurea and mifepristone. The high expression of Th1 (TNF-α, IFN-γ) may cause abortion and be harmful to pregnancy. Th2 type (IL-4, IL-10) may facilitate pregnancy. The expression co-stimulating factor CD80, CD86, CD28, CTLA-4 may be relevant to the spontaneous abortion.
RESUMEN
The kinetics of cytokine mRNA expression was studied in porcine alveolar macrophages using an RT-PCR assay. The expression levels of IFN- gamma, IL-2, IL-4, IL-6, GM-CSF, IL-12 p35, and IL-12 p40 were examined after 2, 4, 14, 24, 48, and 72 h of incubation in unstimulated control and LPS-stimulated cells. The expression contents of IFN-gamma, IL-2, and IL-4 were not detected in both unstimulated and LPS-stimulated cells. On the other hand, the expression levels of IL-6, GM-CSF, and IL-12 in LPS-stimulated cells were almost always higher than those in control cells. Among those cytokines, IL-6 exhibited the predominant expression, and GM-CSF, IL-12 p40, and IL-12 p35 followed in the descending order. The times to reach the peak expression levels for IL-6, and GM-CSF, IL-12 p35, and IL-12 p40 were 14, and 24 h, respectively. After reaching the peak expression point, the expression levels of IL-6, GM-CSF, and IL-12 p40 reduced to the baseline by 72 h after stimulation, however, IL-12 p35 still kept a substantial expression by the same time. This study demonstrates that porcine alveolar macrophages primarily respond to express IL-6, GM-CSF, and IL-12 by LPS-stimulation and have a cytokine-specific expression profile during the stimulation time.
Asunto(s)
Animales , Femenino , Células Cultivadas , Citocinas/genética , Perfilación de la Expresión Génica , Regulación de la Expresión Génica/efectos de los fármacos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Interferón gamma/genética , Interleucinas/genética , Cinética , Lipopolisacáridos/farmacología , Macrófagos Alveolares/efectos de los fármacos , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sensibilidad y Especificidad , Porcinos/genéticaRESUMEN
Objective:To investigate the cytokine profile expressed by CIK cells.Methods:CIK cells were induced from peripheral blood mononuclear cells in the presence of IFN-?,IL-1?,IL-2 and mAb against CD3. The phenotype and characterization of CIK cells were identified by flow cytometric analysis.Cytokine expression profiles were determined at mRNA level by semi-quantitative RT-PCR.Results:The percentage of CD3~+CD56~+ positive cells reached up to or higher than 50%.CIK cells couldn't express IFN-?、IFN-? and IFN-? under this kind of condition. But they could express IFNs principally on day 10 to 20. The level of ILs went down after steadily expressed on day 6 to 10(or 20).TNF-? and TRAIL constitutively expressed during the culture period in vitro. TNF-? maintained high expression on day 20.TGF-?_1 mainly expressed on day 30.Conclusion:CIK cells can widely express various kinds of cytokines in vitro.