RESUMO
The aim of the current study was to examine the protective effects and mechanisms of Ndfipl on neurocytes in an experimental in vitro Parkinson's disease model induced by MPP+. The cell model was developed with dominant negative expression and suppressed expression of Ndfipl by means of transient transfection of Ndfipl-dominant negative and -inhibitory vectors. In total, four different Ndfipl cell models were established. Different methods were used to analyze the cells. The MTT method was used to detect the effect of Ndfipl on the survival rate and apoptosis of the cells induced by MPP(+). We further studied the roles of Ndfipl in inhibiting MPP(+)-induced SH-SY5Y apoptosis, protection, and ubiquitination of SH-SY5Y cells. Our results showed that Ndfipl reduced apoptosis and improved cell survival rate, indicating that Ndfipl has a neuroprotective effect. Furthermore, we found that Ndfipl binds to Nedd4-1, and that increased expression of Ndfipl significantly reduced Itch expression. We also found that increased ubiquitination played a role in Ndfipl-mediated processes, and that Ndfipl and α-synuclein interact. Additionally, the expression of Ndfipl reduced expression of α-synuclein. In conclusion, Ndfipl plays a significant role in protecting SH-SY5Y cells in in vitro Parkinson's disease models.
Assuntos
Apoptose/genética , Proteínas de Transporte/biossíntese , Complexos Endossomais de Distribuição Requeridos para Transporte/genética , Proteínas de Membrana/biossíntese , Doença de Parkinson Secundária/genética , Ubiquitina-Proteína Ligases/genética , alfa-Sinucleína/biossíntese , 1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/efeitos adversos , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Complexos Endossomais de Distribuição Requeridos para Transporte/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Células HEK293 , Humanos , Intoxicação por MPTP , Ubiquitina-Proteína Ligases Nedd4 , Neurônios/metabolismo , Neurônios/patologia , Fármacos Neuroprotetores , Doença de Parkinson Secundária/induzido quimicamente , Doença de Parkinson Secundária/patologia , Espécies Reativas de Oxigênio/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , alfa-Sinucleína/genéticaRESUMO
This study investigated the effects induced by co-culturing human primary basic fibroblasts (HPBFs) with 16-human bronchial epithelial cells (16-HBE) infected with respiratory syncytial virus (RSV), in particular the transformation of HPBFs into myofibroblasts and secretion of extracellular matrix proteins. HPBFs were co-cultured with 16-HBE cells infected with RSV and quantitatively analyzed. We constructed models of HPBFs co-cultured with 16-HBE cells that were either uninfected (control group) or infected with RSV (experimental group). Following initiation of co-cultures, HPBFs and supernatants were collected at 24-h intervals up to 120 h. Expression of α-smooth muscle actin (α-SMA) was detected by indirect immunofluorescence and western blotting, while type I collagen (Col I) and fibronectin were analyzed by competitive enzyme-linked immunosorbent assays. After 72 h, α-SMA expression increased in HPBFs cultured with RSV-infected 16-HBE relative to uninfected controls, reaching its highest level at 96 h. Similarly, Col I secretion was also higher in HPBFs co-cultured with RSV-infected 16-HBE relative to uninfected controls; Col I secretion increased with time and reached its highest level at 120 h. HPBFs were transformed into myofibroblasts following co-culture with RSV-infected 16-HBE, which when combined with the observed increase in Col I secretion suggests that airway remodeling would then be promoted.
Assuntos
Remodelação das Vias Aéreas , Células Epiteliais/virologia , Fibroblastos/patologia , Miofibroblastos/patologia , Infecções por Vírus Respiratório Sincicial/patologia , Infecções Respiratórias/patologia , Actinas/metabolismo , Brônquios/patologia , Brônquios/virologia , Técnicas de Cocultura , Colágeno Tipo I/metabolismo , Células Epiteliais/patologia , Fibroblastos/metabolismo , Fibronectinas/metabolismo , Humanos , Miofibroblastos/metabolismo , Infecções por Vírus Respiratório Sincicial/metabolismo , Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sinciciais Respiratórios , Infecções Respiratórias/metabolismo , Infecções Respiratórias/virologiaRESUMO
We investigated protein expression in the medullary visceral zone (MVZ) of rats with multiple-organ dysfunction syndrome (MODS) caused by subarachnoid hemorrhage (SAH) to discuss the possible regulatory mechanism of the MVZ in the course of SAH-induced MODS. A SAH-induced MODS model was established in rats by injecting arterial blood into the Willis' circle. Protein expression in the MVZ was analyzed by immunohistochemistry assay. Protein expression in the MVZ peaked 24-36 h after SAH, and was significantly higher than in the control and sham operation groups. Organs at each time point exhibited inflammatory injuries to varying degrees after SAH, which reached a maximum at 24-36 h. Incidences of systemic inflammatory response syndrome and MODS were 100 and 71.67%, respectively, after SAH. There is a consistency between MVZ protein expression and inflammatory changes in each organ after SAH. This prompts the suggestion that the MVZ may be one of the direct regulative centers in SAH-induced MODS, and may be involved in the functional regulation of the surrounding organs after SAH.
Assuntos
Bulbo/metabolismo , Hemorragia Subaracnóidea/metabolismo , Animais , Lesões Encefálicas/metabolismo , Estudos de Casos e Controles , Círculo Arterial do Cérebro/metabolismo , Modelos Animais de Doenças , Imuno-Histoquímica , Masculino , Insuficiência de Múltiplos Órgãos/sangue , Insuficiência de Múltiplos Órgãos/metabolismo , Ratos , Ratos Wistar , Hemorragia Subaracnóidea/sangueRESUMO
This study aimed to isolate mesenchymal stem cells from bone mesenchymal stem cells (BMSCs), determine their therapeutic potential for treating rats with acute liver failure (ALF), further explore the factors that induce liver failure mechanisms, and elucidate the role of bone marrow stem cell therapy and BMSCs on liver homing. We found that differentiation potential was present in BMSCs expressing high levels of CD29 and CD90. These cells improved liver functioning in vivo after transplantation into rat livers with D-galactosamine damage, as evidenced by the levels of alanine aminotransferase and aspartate aminotransferase returning to normal (low levels) in recipient ALF rats. A significant improvement in the liver functional test and histological findings was observed in the transplantation group after 120 and 168 h of transplantation (P < 0.05). Histological data revealed that hepatocyte cell apoptosis was lower in the transplantation group compared to the control groups (P < 0.05), and that the transplantation of BMSCs reduced liver inflammation, decreased hepatic denaturation and necrosis, and promoted liver regeneration. These ameliorations were not recorded in the control groups. The results of in situ hybridization, immunofluorescence staining, and Western blot confirmed the presence of transplanted BMSCs in recipient rat livers. Stromal cell derived factor-1 alpha and vascular endothelial growth factor were significantly upregulated after the intraportal transplantation of BMSCs, with significantly higher levels being found in the portal vein and the tail vein groups (P < 0.05). In conclusion, BMSCs have a therapeutic effect against ALF rats, evoke endogenous repair mechanisms in the liver, and may represent a novel form of therapeutic intervention for the disease. Furthermore, intraportal transplantation serves as a more effective pathway compared to tail vein transplantation.