RESUMO
Hepatitis E virus (HEV) is a foodborne zoonotic pathogen that is supposed to be one of the most common causes of acute viral hepatitis. However, HEV infection has been recently associated with a wide spectrum of extrahepatic manifestations, particularly neurological disorders. Previous studies have shown that HEV is able to cross the blood-brain barrier (BBB) and induce inflammatory response of the central nervous system. However, the pathogenesis of HEV-induced neuroinflammation and tissue injury of the central nervous system have yet to be fully elucidated. In this study, activation of NLRP3 inflammasome following HEV infection were investigated. In a gerbil model infected by HEV, brain histopathological changes including gliosis, neuronophagia and neuron injury were observed and expression of NLRP3, caspase-1, IL-1ß and IL-18 were elevated. Brain microvascular endothelial cells (BMECs) are key components of the BBB that protects the brain from various challenges. Following HEV infection, virus-like particles range from 30 to 40 nm in diameter were observed in human BMECs (hBMECs). Enhanced expression levels of NLRP3 and subsequent ASC, caspase-1, IL-1ß and IL-18 were detected in infected cells. Treatment with MCC950 alleviated HEV infection induced activation of NLRP3 inflammasome, mitochondrial damage and VE-cadherin degradation. The findings provide new insights into HEV-associated neuroinflammation. Moreover, targeting NLRP3 inflammasome signalling is a promising therapeutic in HEV-induced neurological disorder.
RESUMO
Porcine circovirus type 3 (PCV3) is an emerging porcine circovirus that has been associated with porcine dermatitis and nephropathy syndrome (PDNS)-like clinical signs, reproductive failure, cardiac pathologies, and multisystemic inflammation in piglets and sows. Many aspects of PCV3 infection biology and pathogenesis, however, remain unknown. Here, we used a PCV3 virus stock from the rescue of an infectious PCV3 DNA clone to intranasally inoculate 4- and 8-week-old specific-pathogen-free piglets for evaluation of PCV3 pathogenesis. For 4-week-old piglets, typical clinical signs resembling those of PDNS-like disease were observed when piglets were inoculated with PCV3 alone or PCV3 combined with immunostimulation by keyhole limpet hemocyanin, with a mortality of 40% (2/5) for both types of inoculated piglets during a 28-day observation period postinoculation. Both types of inoculated piglets showed similar progressive increases in viral loads in the sera and had seroconverted to PCV3 capsid antibody after inoculation. Pathological lesions and PCV3-specific antigen were detected in various tissues and organs, including the lung, heart, kidney, lymph nodes, spleen, liver, and small intestine, in both types of inoculated piglets. The levels of proinflammatory cytokines and chemokines, including interleukin 1 beta (IL-1ß), IL-6, IL-23α, gamma interferon (IFN-γ), tumor necrosis factor alpha (TNF-α), and chemokine ligand 5 (CCL5), were significantly upregulated in both groups of inoculated piglets. Eight-week-old piglets also exhibited a similar PDNS-like disease but without death after PCV3 inoculation, as evidenced by pathological lesions and PCV3 antigen in various tissues and organs. These results show for the first time successful reproduction of PDNS-like disease by PCV3 infection and further provide significant information regarding the pathogenesis of PCV3 in piglets.IMPORTANCE Porcine circovirus type 3 (PCV3), an emerging porcine circovirus, is considered the cause of porcine dermatitis and nephropathy syndrome (PDNS)-like clinical signs and other systemic diseases in piglets and sows. To evaluate the pathogenesis of PCV3 infection in vivo, we used a PCV3 virus stock from the rescue of an infectious PCV3 DNA clone to intranasally inoculate 4- and 8-week-old specific-pathogen-free piglets and demonstrated successful reproduction of PDNS-like disease in animals that were inoculated with PCV3 alone or PCV3 combined with immunostimulation by keyhole limpet hemocyanin. Both 4- and 8-week-old PCV3-inoculated piglets showed similar increases in viral loads in the sera and had seroconverted to PCV3 capsid antibody. Pathological lesions and PCV3-specific antigen were detected in various tissues and organs, while numerous proinflammatory cytokines and chemokines in the sera were significantly upregulated after PCV3 inoculation. These results will provide significant information regarding the pathogenesis of PCV3 in piglets.
Assuntos
Circovirus/metabolismo , Dermatite/metabolismo , Doenças dos Suínos/virologia , Animais , Anticorpos Antivirais/sangue , Antígenos Virais/imunologia , Capsídeo/imunologia , Proteínas do Capsídeo/imunologia , Infecções por Circoviridae/virologia , Circovirus/genética , Dermatite/virologia , Genoma Viral/genética , Rim/patologia , Fígado/patologia , Pulmão/patologia , Pulmão/virologia , Suínos/virologiaRESUMO
With the globe warming, chronic heat stress (CHS) has been considered to be a common hazard that could negatively affect pig's growth and reproduction performance. However, the effects of CHS on the immune functions of pigs were seldom reported, especially the cellular immune functions of intestinal mucosal system. In order to resolve this problem, a pig CHS model was built firstly and the effects of CHS on numbers of T cells in spleen and small intestines were observed. Exposure to a temperature of 39⯰C, 4â¯h/d for 10d, the expression of heat stress protein 70 (HSP70) was increased dramatically. Under CHS condition, the numbers of CD3+ T cells were increased dramatically in both spleens and small intestines. Besides, the numbers of CD4+T cells and the value of CD4+/CD8+T cells in spleens were also significantly increased. The results highly revealed that CHS made the equilibrium state of immune function destroyed. Furthermore, CHS mainly promoted the expression of anti-apoptosis factor B cell lymphoma-2 (Bcl-2) and thus inhibited the apoptosis of lymphocytes in spleens and intestinal mucosa. This study demonstrates for the first time that CHS negatively affects the immune functions of both spleens and intestinal mucosal system in pigs through the inhibition of apoptosis. Our study can richer the data for study of mechanism of CHS and provide new knowledge for reference of making new strategy to control the disease induced by CHS.
Assuntos
Apoptose/efeitos da radiação , Resposta ao Choque Térmico , Mucosa Intestinal/imunologia , Mucosa Intestinal/efeitos da radiação , Baço/imunologia , Baço/efeitos da radiação , Animais , Temperatura Alta , Suínos , Fatores de TempoRESUMO
The health effects of bisphenol A (BPA) have become a great concern in recent years. In this study, the reproductive toxicity of BPA was investigated. Male CD-1 mice were orally administrated with BPA (0, 100, 300 and 600 mg kg-1 body weight) for 56 consecutive days. Results showed that relative testis weight to total body weight was significantly lower in the high-dose group ( p < 0.01, p < 0.05). Microscopic examination under light and transmission electron microscopes showed disorders of spermatogenesis after BPA exposure, including rough basal lamina of seminiferous tubules and damage of tight junctions between Sertoli cells. Further study by terminal-deoxynucleoitidyl transferase-mediated nick end labelling assay showed a significant induction of apoptosis in the testis tissue of the BPA groups ( p < 0.01). Immunohistochemical study found that the expression of androgen-binding protein (ABP) was significantly decreased in BPA-treated mice ( p < 0.01). Our results indicated that impairment of the basal lamina of seminiferous tubules and tight junctions may contribute to BPA-induced cell injury. A decrease in the level of ABP could be the possible mechanism for the reproductive toxicity of BPA. These findings provided direct evidence and novel insight into the reproductive toxicity of BPA and may have implications for understanding the toxicity of other endocrine disruptors.
Assuntos
Compostos Benzidrílicos/toxicidade , Fenóis/toxicidade , Testículo/efeitos dos fármacos , Animais , Apoptose , Disruptores Endócrinos/toxicidade , Masculino , Camundongos , Reprodução/efeitos dos fármacos , Túbulos Seminíferos/efeitos dos fármacos , Túbulos Seminíferos/patologia , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/patologia , Espermatogênese/efeitos dos fármacos , Testículo/patologia , Junções Íntimas/efeitos dos fármacosRESUMO
It has been shown that enhancement of blood-brain barrier (BBB) permeability is modulated by the expression of chemokines/cytokines and reduction of tight junction (TJ) proteins in the brains of mice infected with rabies virus (RABV). Since CXCL10 was found to be the most highly expressed chemokine, its temporal and spatial expression were determined in the present study. The expression of the chemokine CXCL10 was initially detected in neurons as early as 3 days postinfection (p.i.) in the brains of RABV-infected mice, after which it was detected in microglia (6 days p.i.) and astrocytes (9 days p.i.). Neutralization of CXCL10 by treatment with anti-CXCL10 antibodies reduced gamma interferon (IFN-γ) production and Th17 cell infiltration, as well as restoring TJ protein expression and BBB integrity. Together, these data suggest that it is the neuronal CXCL10 that initiates the cascade that leads to the activation of microglia/astrocytes, infiltration of inflammatory cells, expression of chemokines/cytokines, reduction of TJ protein expression, and enhancement of the BBB permeability.
Assuntos
Barreira Hematoencefálica/fisiopatologia , Quimiocina CXCL10/biossíntese , Citocinas/metabolismo , Leucócitos/imunologia , Neurônios/virologia , Vírus da Raiva/imunologia , Raiva/imunologia , Animais , Feminino , Perfilação da Expressão Gênica , Camundongos Endogâmicos ICR , Neurônios/imunologia , Permeabilidade , Vírus da Raiva/fisiologia , Proteínas de Junções Íntimas/biossíntese , Fatores de TempoRESUMO
BACKGROUND: Hemoglobin is a rich source of biological peptides. As a byproduct and even wastewater of poultry-slaughtering facilities, chicken blood is one of the most abundant source of hemoglobin. RESULTS: In this study, the chicken hemoglobin antimicrobial peptides (CHAP) were isolated and the antimicrobial and bactericidal activities were tested by the agarose diffusion assay, minimum inhibitory concentration (MIC) analysis, minimal bactericidal concentration (MBC) analysis, and time-dependent inhibitory and bactericidal assays. The results demonstrated that CHAP had potent and rapid antimicrobial activity against 19 bacterial strains, including 9 multidrug-resistant bacterial strains. Bacterial biofilm and NaCl permeability assays, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were further performed to detect the mechanism of its antimicrobial effect. Additionally, CHAP showed low hemolytic activity, embryo toxicity, and high stability in different temperatures and animal plasma. CONCLUSION: CHAP may have great potential for expanding production and development value in animal medication, the breeding industry and environment protection.
Assuntos
Anti-Infecciosos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Hemoglobinas/farmacologia , Fragmentos de Peptídeos/farmacologia , Animais , Antibacterianos/efeitos adversos , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Anti-Infecciosos/isolamento & purificação , Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Bactérias/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Bacteriólise/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Atividade Bactericida do Sangue , Permeabilidade da Membrana Celular/efeitos dos fármacos , Galinhas , Farmacorresistência Bacteriana Múltipla , Hemoglobinas/química , Hemolíticos/isolamento & purificação , Hemolíticos/farmacologia , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Fragmentos de Peptídeos/química , Estabilidade Proteica , Cloreto de Sódio/metabolismoRESUMO
BACKGROUND: In recent decades, Porcine circovirus 2 (PCV2) infection has been recognized as the causative agent of postweaning multisystemic wasting syndrome, and has become a threat to the swine industry. Hepatitis E virus (HEV) is another high prevalent pathogen in swine in many regions of the world. PCV2 and HEV are both highly prevalent in pig farms in China. CASE PRESENTATION: In this study, we characterized the HEV and PCV2 co-infection in 2-3 month-old piglets, based on pathogen identification and the pathological changes observed, in Hebei Province, China. The pathological changes were severe, and general hyperemia, hemorrhage, inflammatory cell infiltration, and necrosis were evident in the tissues of dead swine. PCR was used to identify the pathogen and we tested for eight viruses (HEV, Porcine reproductive and respiratory syndrome virus, PCV2, Classical swine fever virus, Porcine epidemic diarrhea virus, Transmissible gastroenteritis coronavirus, Porcine parvovirus and Pseudorabies virus) that are prevalent in Chinese pig farms. The livers, kidneys, spleens, and other organs of the necropsied swine were positive for HEV and/or PCV2. Immunohistochemical staining showed HEV- and PCV2-antigen-positive signals in the livers, kidneys, lungs, lymph nodes, and intestine. CONCLUSION: HEV and PCV2 co-infection in piglets was detected in four out of seven dead pigs from two pig farms in Hebei, China, producing severe pathological changes. The natural co-infection of HEV and PCV2 in pigs in China has rarely been reported. We speculate that co-infection with PCV2 and HEV may bring some negative effect on pig production and recommend that more attention should be paid to this phenomenon.
Assuntos
Infecções por Circoviridae/veterinária , Circovirus , Vírus da Hepatite E , Hepatite E/veterinária , Doenças dos Suínos/virologia , Animais , China/epidemiologia , Infecções por Circoviridae/complicações , Infecções por Circoviridae/epidemiologia , Infecções por Circoviridae/patologia , Circovirus/genética , Coinfecção/patologia , Coinfecção/veterinária , Coinfecção/virologia , Hepatite E/complicações , Hepatite E/epidemiologia , Hepatite E/patologia , Vírus da Hepatite E/genética , Filogenia , Reação em Cadeia da Polimerase/veterinária , Suínos/virologia , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/patologiaRESUMO
Bisphenol A (BPA) is a chemical estrogen widely used in the food packaging industry, especially in baby bottles. Its toxicity for the fetus has become a great concern in recent years. In the present study, the effects of BPA on the development of central immune organs in chick embryos were investigated. A total of 30 specific-pathogen-free (SPF) chick embryos were divided into BPA, control, and vehicle group. Chick embryos were exposed to BPA (250 µg per egg), saline (control), or corn oil (vehicle) on embryonic day 9 (ED9) by injection into the allantoic cavity. Thymuses and bursae of Fabricius were collected on ED22. The microscopic examination of tissue structure and ultrastructure was carried out for histopathological changes of thymus and the bursa of Fabricius morphology under light and scanning electron microscopes (SEM). In the BPA group, the weight index of the bursae of Fabricius was significantly reduced (p < 0.01); the number of lymphatic follicles in the bursae of Fabricius was remarkably decreased (p < 0.01); and the thickness of the thymus cortex and medulla was reduced (p < 0.01). Light microscope and SEM examinations further showed that the lymphatic follicles and epithelial cells of the bursa of Fabricius and thymus were damaged by BPA. Our study confirms a direct toxicity of BPA at a very low-dose level on the development of the central immune organs of SPF chick embryos. However, more studies are necessary to elucidate the underlying mechanisms.
Assuntos
Compostos Benzidrílicos/toxicidade , Bolsa de Fabricius/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Fenóis/toxicidade , Timo/efeitos dos fármacos , Animais , Bolsa de Fabricius/patologia , Embrião de Galinha , Tamanho do Órgão/efeitos dos fármacos , Organismos Livres de Patógenos Específicos , Timo/citologia , Timo/patologiaRESUMO
Previous studies here have demonstrated that the rabbit sacculus rotundus-derived antimicrobial peptides (RSRP) could alter the intestinal mucosal immune responses in specific-pathogen-free (SPF) chickens, however, the protective effects of RSRP on chickens against infection remain questionable. In the present study, eighty SPF chickens were randomly divided into five groups and challenged with very virulent infectious bursal disease virus (vvIBDV) to determine the protective effects and its underlying mechanism of RSRP. Histopathology examination found that vvIBDV-infection caused severe damage in the bursa of Fabricius, especially the bursal lymphoid follicles underwent severe necrosis, depletion, hemorrhage, and edema. Unexpectedly, RSRP intervention significantly reduced the necrosis and depletion of lymphoid follicles in the vvIBDV-infected chickens. Moreover, RSRP treatment significantly decreased the expression of Bax (P < 0.01) as well as remarkably promoted the expression of Bcl-2 (P < 0.01), concomitantly alleviated the excessive apoptosis in the immune organs such as the bursa of Fabricius during vvIBDV infection. Notably, consistent with our previous reports that increased mast cell activation and degranulation in the bursa after vvIBDV infection, RSRP administration considerably reduced the mast cell density and the expression of tryptase, a marker for activated mast cells. Collectively, the present study indicates that rabbit sacculus rotundus-derived antimicrobial peptides could effectively protect the major immune organs including the bursa of Fabricius from the damage caused by vvIBDV infection, which provides the possibility and a promising perspective for the future application of antimicrobial peptides for poultry production.
Assuntos
Infecções por Birnaviridae , Galinhas , Vírus da Doença Infecciosa da Bursa , Doenças das Aves Domésticas , Animais , Infecções por Birnaviridae/veterinária , Infecções por Birnaviridae/virologia , Infecções por Birnaviridae/prevenção & controle , Vírus da Doença Infecciosa da Bursa/fisiologia , Doenças das Aves Domésticas/virologia , Doenças das Aves Domésticas/prevenção & controle , Doenças das Aves Domésticas/imunologia , Coelhos , Organismos Livres de Patógenos Específicos , Bolsa de Fabricius/efeitos dos fármacos , Bolsa de Fabricius/virologia , Peptídeos Antimicrobianos/farmacologia , Peptídeos Antimicrobianos/administração & dosagem , Distribuição AleatóriaRESUMO
Avian metapneumovirus causes acute respiratory tract infection and reductions in egg production in various avian species. We isolated and characterized an increasingly prevalent avian metapneumovirus subgroup C strain from meat-type commercial chickens with severe respiratory signs in China. Culling of infected flocks could lead to economic consequences.
Assuntos
Galinhas/virologia , Metapneumovirus/genética , Infecções por Paramyxoviridae/veterinária , Doenças das Aves Domésticas/epidemiologia , Infecções Respiratórias/veterinária , Animais , China/epidemiologia , Genes Virais , Pulmão/patologia , Pulmão/virologia , Tipagem Molecular , Infecções por Paramyxoviridae/epidemiologia , Infecções por Paramyxoviridae/virologia , Filogenia , Doenças das Aves Domésticas/virologia , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Análise de Sequência de DNARESUMO
BACKGROUND: Using various methods, we analyzed the cause of death among weaned pigs from a pig farm in Hebei Province, China. All 300 piglets (100% fatality) were identified as moribund, with death occurring within 1 month from the onset of clinical signs. RESULTS: A single case exhibited obvious hemorrhagic necrotic changes with massive lymphocytic infiltration in multiple organs, in particular the liver, lungs and intestines. Dysplasia and lymphocyte deterioration were common in lymphatic organs. No visible bacterial colonies from liver and spleen were observed in nutrient, MacConkey, and blood agar plates. Using polymerase chain reaction techniques for this case, we attempted to detect a number of epidemic swine viruses in spleen and liver, including PRRSV, CSF, HEV, and PCV2. We found that this sample was positive for the presence of HEV and PRRSV. CONCLUSIONS: We have detected HEV and PRRSV co-infection in one piglet. Severe pathologic changes were observed. The high mortality of weaned pigs which showed the similar clinical syptom was possibly a result of HEV and PRRSV co-infection, which has rarely been reported previously. We speculated that co-infection with PRRSV and HEV might lead to more serious problems.
Assuntos
Coinfecção/veterinária , Vírus da Hepatite E/isolamento & purificação , Hepatite E/veterinária , Síndrome Respiratória e Reprodutiva Suína/diagnóstico , Vírus da Síndrome Respiratória e Reprodutiva Suína/isolamento & purificação , Estruturas Animais/patologia , Animais , China , Coinfecção/diagnóstico , Coinfecção/patologia , Coinfecção/virologia , Infecções por Coronaviridae , Hepatite E/complicações , Hepatite E/diagnóstico , Hepatite E/patologia , Reação em Cadeia da Polimerase , Síndrome Respiratória e Reprodutiva Suína/patologia , Síndrome Respiratória e Reprodutiva Suína/virologia , SuínosRESUMO
Newcastle disease, which is caused by Newcastle disease virus (NDV), is a highly contagious viral disease of poultry and other bird species. The mucosa is the first line of defence to invading pathogens, including NDV, and it has been confirmed that the mucosa can contribute to host protection. This study was conducted to evaluate the intestinal mucosal immunology in NDV infection. Forty specific-pathogen-free chickens were divided into two groups, 20 birds in each group. Group 1 was inoculated with NDV by the intravenous route. Group 2 was used as the control group and was given sterile phosphate-buffered saline by the same route. At 24, 48, 72, and 96 h post infection (h.p.i.), five chickens from each treatment were killed. Samples of the duodenum, jejunum, and ileum were collected to quantify intestinal intraepithelial lymphocytes (IEL), goblet cells and secretory IgA (sIgA) by cytochemistry and immunohistochemistry analysis. The results indicated that IEL were increased from 24 to 72 h.p.i. in the infected tissues, and were significantly higher than in the control group at 48 h.p.i. (P < 0.01). In contrast to IEL, goblet cell numbers were reduced dramatically from 24 to 96 h.p.i. in the infected birds (P < 0.01) Furthermore, the content of sIgA was significantly higher at 48 and 72 h.p.i. in the infected tissues (P < 0.01). sIgA positivity was observed in the epithelial lining of the intestinal mucosa. These data suggest that IEL, goblet cells, and sIgA were involved in the intestinal mucosal immunity against NDV infection.
Assuntos
Galinhas , Células Caliciformes/imunologia , Mucosa Intestinal/imunologia , Linfócitos/imunologia , Doença de Newcastle/imunologia , Doença de Newcastle/virologia , Vírus da Doença de Newcastle/imunologia , Animais , Imunoglobulina A Secretora/metabolismo , Imuno-Histoquímica/veterinária , Mucosa Intestinal/metabolismoRESUMO
Hepatitis B virus (HBV) is one of the most important human pathogens. Its existence in food animals could present a significant threat to public health. The objective of this study was to determine if HBV is present in serum and liver of chickens. A total of 129 serum samples from broiler chickens were collected for the detection of HBV antigens and antibodies, and 193 liver samples were tested for HBV DNA sequence by PCR and for the existence of HBV antigens by immunohistochemistry. The overall prevalence of HBsAg, anti-HBs, anti-HBc was 28.68%, 53.49%, 17.05%, respectively, whereas HBeAg, anti-HBe were barely detectable. Three serum samples were found to be positive for both HBsAg and HBeAg. Further analysis of these samples with transmission electron microscopy (TEM) revealed two morphologic particles with 20 nm and 40 nm in diameter, which were similar to small spherical and Danes particles of HBV. The viral DNA sequence identified in two of the chicken livers shared 92.2% of one known HBV strain and 97.9% nucleotide sequence of another HBV strain. Our results showed the existence of HBV in chickens. This would present a significant risk to people who work with live chickens or chicken products if HBV found in chicken could be confirmed to be the same as human HBV.
Assuntos
Galinhas/virologia , Vírus da Hepatite B/isolamento & purificação , Hepatite B/veterinária , Doenças das Aves Domésticas/virologia , Animais , Sequência de Bases , DNA Viral/química , Hepatite B/epidemiologia , Hepatite B/virologia , Anticorpos Anti-Hepatite B/sangue , Antígenos do Núcleo do Vírus da Hepatite B/sangue , Antígenos de Superfície da Hepatite B/sangue , Antígenos E da Hepatite B/sangue , Vírus da Hepatite B/imunologia , Vírus da Hepatite B/ultraestrutura , Humanos , Fígado/imunologia , Fígado/virologia , Dados de Sequência Molecular , Doenças das Aves Domésticas/epidemiologia , Alinhamento de SequênciaRESUMO
The 3-methyl-4-nitrophenol (4-nitro-m-cresol; PNMC) exists in diesel exhaust particles (DEP), and is also one of the degradation products of insecticide fenitrothion. To assess potential nephrotoxicity of PNMC, male Sprague-Dawley (SD) rats were subcutaneously dosed with PNMC at 1, 10, and 100 mg/kg/day for five consecutive days. No significant changes were detected in body weights and relative weights of kidneys by the treatment of PNMC. However, the extent of cellular necrosis was found to be severe in renal tubular epithelial cells of PNMC-treated rats. In addition, PNMC exposure significantly increased the number of terminal deoxynucleotidyle transferase-mediated dUTP nick end-labeling (TUNEL)-positive cells compared to the control in renal tubule of PNMC-treated rats. Moreover, immunohistochemical results indicated that significant decrease in the B-cell lymphoma 2 (Bcl-2) expressions andincrease in the Bcl-2 associated × protein (Bax) expression were detected in PNMC-treated rats. The ratio of Bcl-2/Bax was also reduced significantly at PNMC-treated rats dosed at 10 or 100 mg kg(-1) . Furthermore, the significant increase of FAS (CD95/APO-1) expression was found in the groups dosed at 10 or 100 mg kg(-1) of PNMC. The expression of Caspase-3 was higher in PNMC-treated rats, compared to the control group. Our results indicated that activation of mitochondria and Caspase-3 protease may contribute to the PNMC-induced apoptosis, suggesting that PNMC could cause both necrosis and apoptosis resulting in cell death of renal epithelium cells and could induce renal toxicity.
Assuntos
Apoptose/efeitos dos fármacos , Cresóis/toxicidade , Células Epiteliais/efeitos dos fármacos , Necrose/induzido quimicamente , Animais , Caspase 3/metabolismo , Células Epiteliais/citologia , Marcação In Situ das Extremidades Cortadas , Túbulos Renais/citologia , Masculino , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos , Ratos Sprague-Dawley , Proteína X Associada a bcl-2/metabolismo , Receptor fas/metabolismoRESUMO
Chronic hepatitis E virus (HEV) infection is frequently reported in immunocompromised patients, but has also been increasingly reported in non-immunocompromised individuals. We characterized the course of chronic HEV infection in immunocompetent rabbits. In two independent experiments, 40 specific-pathogen-free rabbits were infected with a rabbit HEV genotype 3 strain in serial diluted titers (108 to 104 copies/mL). Serum and fecal samples were collected weekly and were tested for HEV RNA, antigen, anti-HEV and liver enzymes. Rabbits that spontaneously cleared the infection before 10 weeks post-inoculation (wpi) were kept to the end of the study as recovery control. Liver tissues were collected from HEV-infected rabbits at 5, 10 and 26 wpi for histopathological analysis. Nineteen rabbits (47.5%) developed chronic HEV infection with persistent viraemia and fecal HEV shedding for >6 months. Seroconversion to anti-HEV was observed in 84.2% (16/19) of the chronically infected rabbits. Serum levels of aminotransferase were persistently elevated in most of the rabbits. Characterizations of chronic HEV infection in immunocompetent settings could be recapitulated in rabbits, which can serve as a valuable tool for future studies on pathogenesis.
Assuntos
Vírus da Hepatite E , Hepatite E , Animais , Vírus da Hepatite E/genética , Humanos , RNA Viral/genética , Coelhos , Organismos Livres de Patógenos Específicos , Eliminação de Partículas ViraisRESUMO
The toxicity of melamine (MA) and its analogue cyanuric acid (CA) in multiple organs has been widely investigated. The purpose of this study was to characterize the pathological lesions of the liver caused by melamine alone or in combination with CA. Mice were oral administered 0, 25, 50, or 100 mg/kg/day MA and CA mixture (MC), or 25, 50, and 100 mg/kg/day MA alone for 7 days. We found MC caused increase of liver weight index and elevations of the serum concentrations of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and creatinine (Cr). Histopathologically, both MA and MC caused scattered necrosis and inflammation cell infiltration in liver. Notably, at 100 mg/kg/day MC, melamine-related crystals were observed in hepatic sinusoid. The liver at high-dose MA and MC groups were further examined by TEM. There were marked degeneration of the mitochondria, and crystal deposition in the Disse space or cytoplasm of hepatic cells and Kupffer cells. TUNEL staining revealed that MA and MC caused apoptosis of hepatocytes and Kupffer cells. Western blotting showed the expression of Bcl-2 decreased, and Bax and caspase-3 increase in liver. The analysis of oxidative stress showed that the expression and activities of two key antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPX) decreased, and the concentration of malondialdehyde (MDA) elevated in MA- and MC-treated mice. These results from this study demonstrated that both MA and MC caused pathological damage to the liver in mice, especially when ingested in high concentration.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/patologia , Triazinas/toxicidade , Animais , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Relação Dose-Resposta a Droga , Fígado/efeitos dos fármacos , Fígado/patologia , Camundongos , Distribuição AleatóriaRESUMO
Recently, hepatitis E virus (HEV) has caused large outbreaks and presented a significant public health problem. Thus, the mechanism of HEV has attracted increasing research attention. Previous studies revealed that HEV infection induced hepatocyte injuries and structural and functional changes in mitochondria. These pathological changes affected the life cycle of hepatocytes. However, the precise underlying mechanism and the effector protein responsible for this process remain unclear. In the present study, mitochondrial function and the expression of mitophagy-associated mRNA transcripts and proteins were detected in an HEV- infected Mongolian gerbil model. Observation of ultrastructural changes in the liver of the inoculated group revealed the disappearance of mitochondrial cristae of mitochondrion, blurring of the bilayer structure and cavitation in the cytoplasm. The results showed that the mitochondrial transmembrane potential of decreased, mitochondrial transition pore (MPTP) opening increased, reactive oxygen species (ROS) production increased, and glutathione peroxidase (GSH-Px) activity decreased in the HEV-inoculated group. Moreover, the LC3, Beclin1, BNIP3L, Parkin, PINK1 and P62 mRNA levels were significantly increased (p < 0.05 and p < 0.01) in the inoculated group. Western blot and immunohistochemistry assay analyses detected the upregulation of the mitophagy-associated proteins LC3, Beclin1, BNIP3L, Parkin, PINK1 and P62 (p < 0.05 and p < 0.01) in HEV-infected gerbils. All these data demonstrated that HEV infection in vivo induced mitochondrial dysfunction and the activation of the mitophagy pathway, which might be one of the key factors in hepatocyte injury.
Assuntos
Vírus da Hepatite E , Animais , Proteína Beclina-1/metabolismo , Gerbillinae/genética , Gerbillinae/metabolismo , Vírus da Hepatite E/genética , Fígado/patologia , Mitocôndrias/metabolismo , Mitofagia , Proteínas Quinases/metabolismo , RNA Mensageiro/metabolismo , Ubiquitina-Proteína Ligases/genéticaRESUMO
BACKGROUND: The objective of this study is to established evidence of the existence of a novel member of the hepadnavirus family endemic in swine. Temporarily this virus was designated as swine hepatitis B virus (SHBV). This SHBV can be detected by using human hepatitis B virus diagnostic kits including ELISA, immunohistochemical staining, and transmission electron microscopy (TEM). Also seroprevalence of pig farms in Beijing, China, and pathological features of SHBV infection was determined. RESULTS: Screened result shows that overall prevalence of HBsAg was 24.8%, closed to that of anti-HBsAg, whereas HBeAg and anti-HBe were barely detectable. The distribution of HBsAg and HBcAg was examined by immunohistochemistry of liver samples. Typical hepatitis pathological change, such as spotty parenchymal cell degeneration, necrosis of hepatocytes and proliferation of fibrous connective tissue were observed during histopathological analysis. Analysis of HBsAg-positive serum with TEM revealed two morphologic forms, 20 nm and 40 nm sized particles, similar to small spherical and Danes particles of HBV. Observation of the ultrastructure of the liver also found HBV-like particles in the nucleus of hepatocytes. CONCLUSION: Our research result implies that SHBV could be a causative agent of swine. The discovery of SHBV will unveil novel evolutionary aspects of hepatitis and provides new information for further hepadnavirus research.
Assuntos
Infecções por Hepadnaviridae/epidemiologia , Hepatite Viral Animal/epidemiologia , Orthohepadnavirus/classificação , Orthohepadnavirus/imunologia , Doenças dos Suínos/epidemiologia , Animais , China/epidemiologia , Doenças Endêmicas , Ensaio de Imunoadsorção Enzimática , Infecções por Hepadnaviridae/virologia , Anticorpos Anti-Hepatite B/sangue , Antígenos de Superfície da Hepatite B/sangue , Antígenos E da Hepatite B/sangue , Hepatite Viral Animal/virologia , Histocitoquímica , Humanos , Imuno-Histoquímica , Fígado/patologia , Fígado/virologia , Microscopia , Microscopia Eletrônica de Transmissão , Orthohepadnavirus/isolamento & purificação , Estudos Soroepidemiológicos , Suínos , Doenças dos Suínos/virologia , Vírion/ultraestruturaRESUMO
Porcine circovirus type 3 (PCV3) infection induces porcine dermatitis and nephropathy syndrome, reproductive failure, and multisystemic inflammatory lesions in piglets and sows. To better understand the host responses to PCV3 infection, isobaric tags for relative and absolute quantification (iTRAQ) labeling combined with LC-MS/MS analysis was used for quantitative determination of differentially regulated cellular proteins in the lungs of specific-pathogen-free piglets after 4 weeks of PCV3 infection. Totally, 3429 proteins were detected in three independent mass spectrometry analyses, of which 242 differential cellular proteins were significantly regulated, consisting of 100 upregulated proteins and 142 downregulated proteins in PCV3-infected group relative to control group. Bioinformatics analysis revealed that these higher or lower abundant proteins involved primarily metabolic processes, innate immune response, MHC-I and MHC-II components, and phagosome pathways. Ten genes encoding differentially regulated proteins were selected for investigation via real-time RT-PCR. The expression levels of six representative proteins, OAS1, Mx1, ISG15, IFIT3, SOD2, and HSP60, were further confirmed by Western blotting and immunohistochemistry. This study attempted for the first time to investigate the protein profile of PCV3-infected piglets using iTRAQ technology; our findings provide valuable information to better understand the mechanisms underlying the host responses to PCV3 infection in piglets. SIGNIFICANCE: Our study identified differentially abundant proteins related to a variety of potential signaling pathways in the lungs of PCV3-infected piglets. These findings provide valuable information to better understand the mechanisms of host responses to PCV3 infection.
Assuntos
Infecções por Circoviridae/metabolismo , Circovirus/metabolismo , Pulmão/metabolismo , Proteoma/metabolismo , Proteômica/métodos , Doenças dos Suínos/metabolismo , Animais , Cromatografia Líquida/métodos , Infecções por Circoviridae/virologia , Circovirus/isolamento & purificação , Circovirus/patogenicidade , Pulmão/virologia , Proteoma/análise , Suínos , Doenças dos Suínos/virologia , Espectrometria de Massas em Tandem/métodosRESUMO
An ideal animal model for hepatitis E virus (HEV) research is still unavailable. To assess the possibility of using Mongolian gerbils as animal model, 28 gerbils were randomly assigned into two groups, 14 for each group. Gerbils in Group 1 were inoculated with a genotype 4 HEV recovered from swine via the intraperitoneal route. Group 2 was used as a negative control and inoculated with normal suspension of swine liver. Sera and feces samples were collected once a week for 7 weeks. Two gerbils from both groups were necropsied weekly, pathological changes were recorded and tissue samples collected for further investigation. Distribution of the virus antigens was determined by immunohistochemical staining. Nested RT-PCR and a commercial ELISA kit were used to confirm the infection. Research results demonstrated that Mongolian gerbils in Group 1 were successfully infected with HEV. Viremia and fecal virus shedding lasted nearly 4 weeks, while the virus could be detected constantly in the liver, and occasionally in the kidneys and spleen as well as the small intestine. Histopathological changes in the liver were present with slight, multifocal, lymphohistiocytic infiltrates in the portal tracts or distributed irregularly throughout the liver. HEV antigens could be detected in the liver and intestine, and were mainly distributed in the nuclei. The results indicate that Mongolian gerbils could be used as an ideal animal model for the study of HEV.