RESUMEN
BACKGROUNDS: (6aS, 10S, 11aR, 11bR, 11cS)-10-methylaminododecahydro-3a, 7a-diaza-benzo (de) anthracene-8-thione (MASM), a novel derivative of matrine, exhibits better anti-inflammatory activity. This study was designed to evaluate the protective effect of MASM on acute and chronic liver injuries and explore the possible mechanisms. METHODS: Acute and chronic liver injury models were established by the CCl4 intraperitoneal injection and the protective effect of MASM was assessed by biochemical and histological examination. The infiltration of different monocyte subsets into the liver was characterized and analyzed by flow cytometry. The in vitro effect of MASM on liver nonparenchymal cells was evaluated by real-time PCR and transwell chemotaxis assays. RESULTS: Administration of MASM markedly attenuated acute liver injury and liver fibrosis induced by CCl4 injection. Meanwhile, the infiltrations of Gr1hi monocytes in injured livers and induced hepatic expression of monocyte chemoattractant protein-1 (MCP-1) were greatly inhibited. Cellular experiments demonstrated that MASM not only decreased the expression of MCP-1 but also inhibited its chemotactic activity. CONCLUSIONS: This study demonstrates that the protective effect of MASM on liver injury could be contributed to the suppression of Gr1hi monocyte infiltration to the liver and the inhibition of MCP-1 production and activity. These findings provide new insights into the protective role of MASM in liver injury.
Asunto(s)
Alcaloides/uso terapéutico , Antracenos/farmacología , Antiinflamatorios/uso terapéutico , Enfermedad Hepática Inducida por Sustancias y Drogas/tratamiento farmacológico , Cirrosis Hepática/tratamiento farmacológico , Monocitos/efectos de los fármacos , Quinolizinas/uso terapéutico , Tionas/farmacología , Alcaloides/farmacología , Animales , Antracenos/uso terapéutico , Antiinflamatorios/farmacología , Antígenos Ly/inmunología , Tetracloruro de Carbono , Enfermedad Hepática Inducida por Sustancias y Drogas/inmunología , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Quimiocina CCL2/inmunología , Hígado/efectos de los fármacos , Hígado/inmunología , Hígado/patología , Cirrosis Hepática/inmunología , Cirrosis Hepática/patología , Masculino , Ratones Endogámicos C57BL , Monocitos/inmunología , Quinolizinas/farmacología , Tionas/uso terapéutico , MatrinasRESUMEN
BACKGROUND: Inula helenium L. is an herb whose anti-inflammatory properties are attributed to its active components, the sesquiterpene lactones (SLs). Our previous study demonstrated that the total sesquiterpene lactones isolated from Inula helenium L. (TSL-IHL), consisting mainly of alantolactone (AL) and isoalantolactone (IAL), may have potential in the prevention and treatment of rheumatoid arthritis (RA). However, the effect of TSL-IHL on atopic dermatitis (AD) has not been studied yet. AIM OF THE STUDY: The present study evaluates the potential of TSL-IHL as a treatment for AD. METHODS/STUDY DESIGNS: The effects of TSL-IHL on the expression of inflammatory genes and the activation of NF-κB signaling pathway in HaCat cells were examined by quantitative real-time PCR and western blotting, respectively, and compared with those of AL and IAL. The protective effect of TSL-IHL against AD was tested in a mouse model induced by 2,4-dinitrochlorobenzene (DNCB), in which AD-like skin lesions were induced in ICR mice by sensitizing once with 100 µl of 7% DNCB painted on their shaved back skin and then challenging with 20 µl of 0.2% DNCB five times on their right ears at 3 day intervals starting on day 5 post-sensitization. RESULTS: TSL-IHL, as well as AL and IAL, could all inhibit TNF-α-induced activation of NF-κB and the expression of TNF-α, IL-1, and IL-4 in HaCat cells in a dose-dependent manner in the range of 0.6-2.4⯵g/ml. The topical application of TSL-IHL (1% W/W in emollient cream) attenuated DNCB-induced dermatitis severity and right ear swelling. The serum levels of IgE, TNF-α and IFN-γ in TSL-IHL-treated mice were reduced by 81.39%, 89.69%, and 87.85%, respectively, while the mRNA levels of IL-4, IL-5 and IL-13, in the back-skin lesions of TSL-IHL-treated mice were reduced by 39.21%, 40.62% and 48.12%, respectively, compared with the untreated controls. Histopathological examination showed that TSL-IHL treatment reduced epidermis/dermis thickening and dermal inflammatory infiltration in both ear and back skins. CONCLUSIONS: We suggest that TSL-IHL inhibited the development of AD-like skin symptoms by regulating cytokine expression and may be an effective alternative therapy for AD.
Asunto(s)
Dermatitis Atópica/tratamiento farmacológico , Inula/química , Lactonas/farmacología , Sesquiterpenos/farmacología , Animales , Línea Celular , Citocinas/metabolismo , Dermatitis Atópica/inducido químicamente , Dinitroclorobenceno , Femenino , Humanos , Ratones , Ratones Endogámicos ICR , FN-kappa B/metabolismo , Extractos Vegetales/farmacología , Piel/efectos de los fármacosRESUMEN
Esculentoside A (EsA), a saponin isolated from Phytolacca esculenta, is reported as a potent suppressor of pro-inflammatory functions of macrophages. However, little is known about the target proteins of EsA for its anti-inflammatory activity. In the present study, to identify the intracellular target for EsA, affinity resins bearing immobilized EsA were used to capture binding proteins of EsA from RAW264.7 cell lysates. Mass spectrography and Western blot analysis of captured proteins indicated that ribosomal protein S3a preferentially bound to EsA affinity resin. Competition experiment further demonstrated that free EsA can disturb the specific interaction between recombinant RPS3a and affinity resin. Surface Plasmon Resonance analysis confirmed that EsA directly bound to RPS3a. Lentivirus-mediated RNAi RPS3a resulted in suppression of TNF-α and IL-6 production and impediment of signal transduction in LPS-stimulated RAW264.7 cells, indicating that RPS3a is required for LPS-triggered signaling during induction of pro-inflammatory cytokines. In addition, EsA inhibited the expression of inflammatory factors more strongly in the case of RPS3a interference. These results suggest that EsA exerts its anti-inflammatory activity by targeting RPS3a and impairing its signaling function. These new findings not only extended our understanding on the intracellular mechanisms of EsA, but also indicated RPS3a as an essential component for LPS-mediated pro-inflammatory signaling, thus implying RPS3a as a novel therapeutic target for anti-inflammatory therapy.
Asunto(s)
Antiinflamatorios/metabolismo , Macrófagos/inmunología , Ácido Oleanólico/análogos & derivados , Proteínas Ribosómicas/metabolismo , Saponinas/metabolismo , Animales , Humanos , Interleucina-6/metabolismo , Lipopolisacáridos/inmunología , Ratones , Ácido Oleanólico/metabolismo , Phytolaccaceae/inmunología , Unión Proteica , Células RAW 264.7 , ARN Interferente Pequeño/genética , Proteínas Ribosómicas/genética , Transducción de Señal , Resonancia por Plasmón de Superficie , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
BACKGROUNDS: Inula helenium L. is an herb with anti-inflammatory properties. Sesquiterpene lactones (SLs), mainly alantolactone (AL) and isoalantolactone (IAL), are considered as its active ingredients. However, the anti-inflammatory effects of SL-containing extracts of I. helenium have not been explored. Here we prepared total SLs from I. helenium (TSL-IHL), analyzed its chemical constituents, and performed cellular and animal studies to evaluate its anti-inflammatory activities. MATERIALS AND METHODS: The chemical profile of TSL-IHL was analyzed by HPLC-UV. Its in vitro effects on the activation of signaling pathways and expression of inflammatory genes were examined by western blotting and quantitative real-time PCR, respectively, and compared with those of AL and IAL. Its in vivo anti-inflammatory effects were evaluated in adjuvant- and collagen-induced arthritis rat models. RESULTS: Chemical analysis showed that AL and IAL represent major constituents of TSL-IHL. TSL-IHL, as well as AL and IAL, could inhibit TNF-α-induced activation of NF-κB and MAPK pathways in b. End3 cells, suppress the expressions of MMP-3, MCP-1, and IL-1 in TNF-α-stimulated synovial fibroblasts, and IL-1, IL-6, and iNOS in LPS-activated RAW 264.7 cells in a dose-dependent manner in the range of 0.6-2.4µg/mL. Oral administration of TSL-IHL at 12.5-50mg/kg could dose-dependently alleviate the arthritic severity and paw swelling in either developing or developed phases of arthritis of rats induced by adjuvant or collagen CONCLUSIONS: These results indicated potentials of TSL-IHL in prevention and therapy of rheumatoid arthritis.
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Antiinflamatorios/uso terapéutico , Artritis Experimental/tratamiento farmacológico , Artritis Reumatoide/tratamiento farmacológico , Inula , Lactonas/uso terapéutico , Sesquiterpenos/uso terapéutico , Animales , Antiinflamatorios/farmacología , Artritis Experimental/metabolismo , Artritis Experimental/prevención & control , Artritis Reumatoide/metabolismo , Artritis Reumatoide/prevención & control , Línea Celular , Citocinas/genética , Citocinas/metabolismo , Lactonas/farmacología , Masculino , Ratones , Proteínas Quinasas Activadas por Mitógenos/metabolismo , FN-kappa B/metabolismo , Raíces de Plantas , Células RAW 264.7 , Ratas Wistar , Sesquiterpenos/farmacologíaRESUMEN
Hepatocytes are critical for the maintenance of liver homeostasis, but its involvement in hepatic fibrogenesis remains elusive. Hepatocyte nuclear factor 1α (HNF1α) is a liver-enriched transcription factor that plays a key role in hepatocyte function. Our previous study revealed a significant inhibitory effect of HNF1α on hepatocellular carcinoma. In this study, we report that the expression of HNF1α is significantly repressed in both human and rat fibrotic liver. Knockdown of HNF1α in the liver significantly aggravates hepatic fibrogenesis in either dimethylnitrosamine (DMN) or bile duct ligation (BDL) model in rats. In contrast, forced expression of HNF1α markedly alleviates hepatic fibrosis. HNF1α regulates the transcriptional expression of SH2 domain-containing phosphatase-1 (SHP-1) via directly binding to SHP-1 promoter in hepatocytes. Inhibition of SHP-1 expression abrogates the anti-fibrotic effect of HNF1α in DMN-treated rats. Moreover, HNF1α repression in primary hepatocytes leads to the activation of NF-κB and JAK/STAT pathways and initiates an inflammatory feedback circuit consisting of HNF1α, SHP-1, STAT3, p65, miR-21 and miR-146a, which sustains the deregulation of HNF1α in hepatocytes. More interestingly, a coordinated crosstalk between hepatocytes and hepatic stellate cells (HSCs) participates in this positive feedback circuit and facilitates the progression of hepatocellular damage. Our findings demonstrate that impaired hepatocytes play an active role in hepatic fibrogenesis. Early intervention of HNF1α-regulated inflammatory feedback loop in hepatocytes may have beneficial effects in the treatment of chronic liver diseases.
Asunto(s)
Células Estrelladas Hepáticas/metabolismo , Factor Nuclear 1-alfa del Hepatocito/biosíntesis , Hepatocitos/metabolismo , Hígado/patología , Animales , Comunicación Celular , Retroalimentación Fisiológica , Fibrosis , Humanos , Quinasas Janus/metabolismo , Hígado/metabolismo , FN-kappa B/metabolismo , Ratas , Factores de Transcripción STAT/metabolismoRESUMEN
To search for novel anticancer agents, we designed and synthesized a series of new triazolyl berberine derivatives. The evaluation of all the synthesized compounds and their anticancer activities against a panel of four human cancer cell lines including MCF-7 (breast), MCF-7/ADR (breast), SW-1990 (pancreatic), SMMC-7721 (liver) and the noncancer cell line HUVEC (human umbilical vein endothelial cell). The results showed that most of the compounds displayed better anticancer activities against MCF-7 and SMMC-7721 compared with berberine. Among these derivatives, compounds 5p and 5a exhibited the most potent inhibitory activities against the SMMC-7721 and SW-1990 cell lines with IC50 values of 14.861 ± 2.4 µM and 16.798 ± 3.4 µM. Furthermore, compounds 5p, 5a and 5n exhibited much better selectivity toward the normal cell line HUVEC than berberine.
Asunto(s)
Antineoplásicos/química , Antineoplásicos/farmacología , Berberina/química , Berberina/farmacología , Línea Celular , Línea Celular Tumoral , Química Clic/métodos , Diseño de Fármacos , Células Endoteliales de la Vena Umbilical Humana , Humanos , Concentración 50 Inhibidora , Células MCF-7 , Relación Estructura-ActividadRESUMEN
To search for novel anticancer agents, we designed and synthesized a series of new triazolyl berberine derivatives. The evaluation of all the synthesized compounds and their anticancer activities against a panel of four human cancer cell lines including MCF-7 (breast), MCF-7/ADR (breast), SW-1990 (pancreatic), SMMC-7721 (liver) and the non-cancer cell line HUVEC (human umbilical vein endothelial cell). The results showed that most of the compounds displayed better anticancer activities against MCF-7 and SMMC-7721 compared with berberine. Among these derivatives, compounds 5p and 5a exhibited the most potent inhibitory activities against the SMMC-7721 and SW-1990 cell lines with IC50 values of 14.861 ± 2.4 µM and 16.798 ± 3.4 µM. Furthermore, compounds 5p, 5a and 5n exhibited much better selectivity toward the normal cell line HUVEC than berberine.
RESUMEN
A series of novel derivatives of phenyl-substituted berberine triazolyls has been designed and synthesized via copper-catalyzed azide-alkyne cycloaddition click chemistry in an attempt to develop antitumor agents. All of the compounds were evaluated for anticancer activity against a panel of three human cancer cell lines, including MCF-7 (breast), SW-1990 (pancreatic), and SMMC-7721 (liver) and the noncancerous human umbilical vein endothelial cell (HUVEC) cell lines. The results indicated that most of the compounds displayed notable anticancer activities against the MCF-7 cells compared with berberine. Among these derivatives, compound 16 showed the most potent inhibitory activity against the SW-1990 and SMMC-7721 cell lines, with half-maximal inhibitory concentration (IC50) values of 8.54±1.97 µM and 11.87±1.83 µM, respectively. Compound 36 exhibited the most potent inhibitory activity against the MCF-7 cell line, with an IC50 value of 12.57±1.96 µM. Compound 16 and compound 36 exhibited low cytotoxicity in the HUVEC cell line, with IC50 values of 25.49±3.24 µM and 30.47±3.47 µM. Furthermore, compounds 14, 15, 16, 17, 18, 32, and 36 exhibited much better selectivity than berberine toward the normal cell line HUVEC.
Asunto(s)
Berberina/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias Hepáticas/tratamiento farmacológico , Neoplasias Pancreáticas/tratamiento farmacológico , Antineoplásicos/síntesis química , Antineoplásicos/química , Antineoplásicos/farmacología , Berberina/síntesis química , Berberina/química , Neoplasias de la Mama/patología , Línea Celular Tumoral , Química Clic/métodos , Diseño de Fármacos , Femenino , Células Endoteliales de la Vena Umbilical Humana , Humanos , Concentración 50 Inhibidora , Neoplasias Hepáticas/patología , Células MCF-7 , Neoplasias Pancreáticas/patología , Relación Estructura-ActividadRESUMEN
UNLABELLED: Liver fibrosis and its endstage, cirrhosis, represent a major public health problem worldwide. Activation of hepatic stellate cells (HSCs) is a central event in hepatic fibrosis. However, the proteins that control HSC activation are incompletely understood. Here we show that (6aS, 10S, 11aR, 11bR, 11cS)-10-methylamino-dodecahydro-3a, 7a-diaza-benzo [de]anthracene-8-thione (MASM) exhibits potent inhibitory activity against liver fibrosis in vitro and in vivo associated with the reduction of Akt phosphorylation. Furthermore, ribosomal protein S5 (RPS5) was identified as a direct target of MASM, which stabilized RPS5 in cultured HSCs and in the liver of experimental animals after dimethylnitrosamine (DMN) or bile duct ligation (BDL). Functional studies revealed that RPS5 could prevent HSC activation. RPS5 overexpression in HSCs resulted in Akt dephosphorylation at both Ser473 and Thr308, and led to subsequent dephosphorylation of GSK3ß or P70S6K. Progression of DMN- and BDL-induced hepatic fibrosis was aggravated by Rps5 knockdown and alleviated by RPS5 overexpression, which correlated with the modulation of Akt phosphorylation and HSC number in the fibrotic livers. Moreover, RPS5 was substantially reduced in the transdifferentiated HSCs, experimental fibrotic livers, and human cirrhosis samples. CONCLUSION: These results demonstrate that RPS5 is implicated in hepatic fibrogenesis and may represent a promising target for potential therapeutic intervention in liver fibrotic diseases.
Asunto(s)
Alcaloides/farmacología , Células Estrelladas Hepáticas/efectos de los fármacos , Células Estrelladas Hepáticas/patología , Compuestos Heterocíclicos de 4 o más Anillos/farmacología , Cirrosis Hepática/tratamiento farmacológico , Cirrosis Hepática/patología , Quinolizinas/farmacología , Proteínas Ribosómicas/fisiología , Adenoviridae/genética , Animales , Línea Celular Transformada , Modelos Animales de Enfermedad , Técnicas de Silenciamiento del Gen , Células Estrelladas Hepáticas/fisiología , Cirrosis Hepática/fisiopatología , Masculino , Medicina Tradicional China/métodos , Miofibroblastos/efectos de los fármacos , Miofibroblastos/patología , Miofibroblastos/fisiología , Fosforilación/efectos de los fármacos , Fosforilación/fisiología , Cultivo Primario de Células , Ratas , Ratas Sprague-Dawley , Proteínas Ribosómicas/genética , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , MatrinasRESUMEN
ZNF24 is a member of the SCAN domain family of Krüppel-like zinc finger (ZF) transcription factors, which plays a critical role in cell proliferation and differentiation. However, how ZNF24 enters the nucleus in order to exert its function remains unclear since its nuclear localization signal(s) (NLS) has not been identified. Here, we generated a series of GFP-tagged deletion and point mutants and assessed their subcellular localization. Our results delimit the NLS to ZF1-2. Deletion of ZF1-2 caused cytoplasmic accumulation of ZNF24. Fusion of the ZF1-2 to green fluorescent protein (GFP) targeted GFP to the nucleus, demonstrating that the ZF1-2 is both necessary and sufficient for nuclear localization. ZNF24 containing histidine to leucine mutations that disrupt the structure of ZF1 or/and ZF2 retains appropriate nuclear localization, indicating that neither the tertiary structure of the zinc fingers nor specific DNA binding are necessary for nuclear localization. K286A and R290A mutation led to partial cytoplasmic accumulation. Co-immunoprecipitation demonstrated that ZNF24 interacted with importin-ß and this interaction required the ZF motifs. The ß-Catenin (CTNNB1) luciferase assays showed that the ZNF24 mutants defective in nuclear localization could not promote CTNNB1 promoter activation as the wild-type ZNF24 did. Taken together, these results suggest that consecutive ZF1-2 is critical for the regulation of ZNF24 nuclear localization and its transactivation function.
Asunto(s)
Núcleo Celular/metabolismo , Factores de Transcripción de Tipo Kruppel/metabolismo , Western Blotting , Línea Celular , Células HeLa , Humanos , Inmunoprecipitación , Factores de Transcripción de Tipo Kruppel/genética , Regiones Promotoras Genéticas/genética , Unión Proteica , Transporte de Proteínas/genética , Transporte de Proteínas/fisiología , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , beta Carioferinas/genética , beta Carioferinas/metabolismoRESUMEN
AIM: To investigate the effects of CpG-oligodeoxynucleotide (CpG-ODN) on the formation of macrophage foam cells and related gene expression. METHODS: A gene expression profile was examined by microarray techniques, and mRNA expression was detected by reverse transcriptase polymerase chain reaction (RT-PCR). The cholesterol and cholesteryl ester contents of cells were determined by high performance liquid chromatography. RESULTS: CD36, LPL, and Fcgamma2b, which were related to lipid metabolism and the formation of macrophage foam cells, were upregulated after CpG-ODN stimulation. The mRNA expression related to the formation of foam cells was confirmed by semiquantitative RT-PCR. Moreover, histochemical analysis confirmed that lipid deposits inside cells increased after CpG-ODN treatment. However, using flow cytometry, we found that CpG-ODN had no effect on the expression of membrane receptors. CONCLUSION: CpG-ODN up-regulated the expression of genes in macrophage foam cell formation.
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Células Espumosas/citología , Oligodesoxirribonucleótidos/farmacología , Adyuvantes Inmunológicos/farmacología , Animales , Antígenos CD36/biosíntesis , Antígenos CD36/genética , Línea Celular , Colesterol/metabolismo , Ésteres del Colesterol/metabolismo , ADN/farmacología , Células Espumosas/metabolismo , Expresión Génica , Humanos , Lipoproteína Lipasa/biosíntesis , Lipoproteína Lipasa/genética , Macrófagos/citología , Macrófagos/metabolismo , Ratones , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Receptores de IgG/biosíntesis , Receptores de IgG/genética , Células U937RESUMEN
The study evaluated the immune response elicited by a DNA vaccine encoding ESAT6 protein of Mycobacterium tuberculosis by DNA prime-protein boost protocol. BALB/c mice were respectively vaccinated with plasmid DNA encoding ESAT6 protein, with ESAT6 protein in IFA adjuvant, or a combined DNA prime-protein boost regimen. While DNA immunization induced Th1-polarized immune response, protein-in-adjuvant vaccination elicited a Th2-dominant response. When animals were primed with DNA and boost with protein, both antibodies and Th-cell proliferative response were significantly enhanced. Moreover, production of Th1-type cytokine (IFN-gamma) was increased significantly by DNA priming-protein boosting. This protocol also resulted in an increased relative ratio of IgG2a to IgG1 and the cytotoxicity of T cells. Thus, this study demonstrated that the formation of ESAT6 DNA prime-protein boost inoculation could improved antigen-specific cellular immune responses, which are important for protection against TB infection.
Asunto(s)
Antígenos Bacterianos/genética , Antígenos Bacterianos/inmunología , Vacunas contra la Tuberculosis/genética , Vacunas contra la Tuberculosis/inmunología , Adyuvantes Inmunológicos , Animales , Anticuerpos Antibacterianos , Proteínas Bacterianas , División Celular/fisiología , Línea Celular , Ensayo de Inmunoadsorción Enzimática , Femenino , Inmunidad Celular , Inmunización Secundaria , Inmunoglobulina G/análisis , Inmunoglobulina G/biosíntesis , Inmunohistoquímica , Interferón gamma/biosíntesis , Interleucina-4/biosíntesis , Linfocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Plásmidos/genética , Bazo/citología , Bazo/inmunología , Células TH1/inmunología , Transfección , Vacunas de ADN/inmunologíaRESUMEN
The immunological protection of pVAX-PS, a DNA vaccine, was assessed in the tree shrews model. pVAX-PS was constructed by inserting the gene encoding the middle (pre-S2 plus S) envelope protein of HBV into a plasmid vector pVAX1. Tree shrews (Tupaia belangeri chinenesis) were experimentally infected with human HBV by inoculation with human serum positive for HBV markers. DNA vaccination-induced seroconversion and antibody to HBV surface antigen (anti-HBs) were analyzed by ELISA, and protective effects elicited by pVAX-PS vaccination against subsequent HBV challenge were evaluated by detection of HBV seromarkers and observation of hepatic lesions in HBV-infected tree shrews. The results shown that anti-HBs were detectable in serum at week 2 after pVAX-PS vaccination and peaked at week 4, and immunization with pVAX-PS decreased the positive conversion rate of HBV seromarkers and relieved hepatic lesions in tree shrews challenged with HBV. These results indicated that pVAX-PS immunization could induce remarkable humoral immune response and prevent the experimental tree shrews from infection of HBV.
Asunto(s)
Vacunas contra Hepatitis B/farmacología , Hepatitis B/prevención & control , Animales , Modelos Animales de Enfermedad , Hepatitis B/inmunología , Hepatitis B/patología , Anticuerpos contra la Hepatitis B/sangre , Vacunas contra Hepatitis B/genética , Humanos , Hígado/patología , Factores de Tiempo , Tupaiidae , Vacunas de ADN/genética , Vacunas de ADN/farmacologíaRESUMEN
The study evaluated DNA vaccination in Taenia solium cysticercosis prevention by using cDNA of an antigen (cC1) from T. solium metacestode. pcDNA3-cC1 DNA vaccine was constructed by inserting the cDNA into the eukaryotic expression plasmid pcDNA3. Positive expression of the pcDNA3-cC1 product was confirmed by its transfection into COS7 cell and enzyme-linked immunoabsorbent assay using serum of pigs infected with T. solium metacestode. Immunisation of BALB/c mice with three injections of pcDNA3-cC1 induced antigen-specific immune responses of the Th1 phenotype. Inoculation of new-born pigs induced protection against challenge with T. solium by 73.3% reduction of the metacestode number. Antibodies elicited by DNA immunisation with pcDNA3-cC1 specifically reacted with native cC1 protein, which was mainly restricted to the cyst wall of T. solium metacestode. Positive apoptosis signals were also detected in the cyst wall cells of metacestode slices from pigs immunised with pcDNA3-cC1 by TUNEL staining method. Those suggested that apoptosis played a role in protecting pigs immunised with pcDNA3-cC1 nucleic acid vaccine from pathogen challenge.