RESUMO
BACKGROUND/AIMS: As a natural antioxidant, verbascoside (VB) is proved to be a promising method for the treatment of oxidative-stress-related neurodegenerative diseases. Thus, this study aimed to investigate the effects of VB on glioblastoma cell proliferation, apoptosis, migration, and invasion as well as the mechanism involving signal transducer and activator of transcription 3 (STAT3) and Src homology 2 domain-containing protein tyrosine phosphatase 1 (SHP-1). METHODS: U87 cells were assigned to different treatments. The MTT assay was used to test cell proliferation, flow cytometry was used to detect cell apoptosis, and a Transwell assay was used for cell migration and invasion. We analyzed the glioblastoma tumor growth in a xenograft mouse model. Western blot analysis was employed to determine the protein expression of related genes. RESULTS: Glioblastoma cells exhibited decreased cell proliferation, migration, invasion, and increased apoptosis when treated with VB or TMZ. Western blot analysis revealed elevated SHP-1 expression and reduced phosphorylated (p)-STAT3 expression in glioblastoma cells treated with VB compared with controls. Correspondingly, in a xenograft mouse model treated with VB, glioblastoma tumor volume and growth were decreased. Glioblastoma xenograft tumors treated with VB showed elevated SHP-1, Bax, cleaved caspase-3, and cleaved PARP expression and reduced p-STAT3, Bcl-2, survivin, MMP-2, and MMP-9 expression. siRNA-SHP-1 inhibited the VB effects on glioblastoma. CONCLUSION: This study demonstrates that VB inhibits glioblastoma cell proliferation, migration, and invasion while promoting apoptosis via SHP-1 activation and inhibition of STAT3 phosphorylation.
Assuntos
Apoptose/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glioblastoma , Glucosídeos/farmacologia , Proteínas de Neoplasias/metabolismo , Fenóis/farmacologia , Proteína Tirosina Fosfatase não Receptora Tipo 6/biossíntese , Fator de Transcrição STAT3/metabolismo , Regulação para Cima/efeitos dos fármacos , Animais , Linhagem Celular Tumoral , Feminino , Glioblastoma/tratamento farmacológico , Glioblastoma/metabolismo , Glioblastoma/patologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Metástase Neoplásica , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Microglia, implicating in such neuro-pathologies as brain inflammation, neurodegeneration, glioma, and neurogenesis, play an important role in central nervous system. Advanced research on microglia is crucial in exploring the neuro-pathology and neuro-physiology of these diseases, so how to culture large numbers of microglia in vitro becomes the base of a research. The wildly used method, at present, obtaining microglia from murine cannot fulfill the requirement of research, costing too much time and needing too many rats. We intend to introduce an optimized method that can harvest large quantities of microglia with high purity. Neonatal 2-3 days old Wistar rats were sacrificed and the cerebral cortices were trypsinized. We primarily cultured mixed cortical cells for 8-10 days. The microglia were harvested from the liquid supernatant; the left cells in the mixed cortical glial culture were passaged at a 1:2 density. After another 8-10 days of culture, microglia were collected again. And then, we passaged the left cells again for acquiring microglia from the third collection. We did not add additional mitogens in the experiment. At last, on average, 7.0 × 10(6) microglia were collected from one neonatal rat. By this modified method, much more microglia can be effectively and easily harvested comparing with the usual protocol before. We compared the characteristics of microglia harvested from these three passages, such as morphology, phenotype, purity, and abilities on proliferation, secretion, and phagocytosis. The cells presented typical microglia morphology, having phenotype markers of CD11b/c and CD45. The microglia from these three passages retained similar phagocytosis and secretion functions. Expanded population of microglia for investigation can be provided by this easy method in a short time with little cost and few rats.
Assuntos
Técnicas de Cultura de Células/economia , Técnicas de Cultura de Células/métodos , Microglia/citologia , Animais , Animais Recém-Nascidos , Proliferação de Células , Separação Celular/economia , Separação Celular/métodos , Células Cultivadas , Análise Custo-Benefício , Eficiência , Citometria de Fluxo/economia , Citometria de Fluxo/métodos , Microglia/fisiologia , Microglia/ultraestrutura , Microscopia de Força Atômica , Microscopia Eletrônica de Varredura , Fagocitose/fisiologia , Ratos , Ratos WistarRESUMO
BACKGROUND: Intracranial bronchogenic cysts (BCs) are extremely rare. To our knowledge, this is the first report of a BC in which lesions involve the middle and posterior cranial fossa, as well as the infratemporal fossa. CASE DESCRIPTION: We present the case of a 38-year-old woman who suffered from a cranial nerve dysfunction for 2 years. Magnetic resonance imaging showed that there were skull base communication lesions across the middle and posterior fossa. The patient was operated on through an infratemporal fossa approach. The final diagnosis was BC after histopathologic examination and immunohistochemical verification. The patient's neurologic dysfunction was partially ameliorated at the half-year follow-up. CONCLUSIONS: Intracranial BCs are rare. However, they should be considered in the differential diagnosis for cystic lesions with edge enhancement or extracranial extension.