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Previous studies have shown that scutellarin inhibits the excessive activation of microglia, reduces neuronal apoptosis, and exerts neuroprotective effects. However, whether scutellarin regulates activated microglia-mediated neuronal apoptosis and its mechanisms remains unclear. This study aimed to investigate whether scutellarin can attenuate PC12 cell apoptosis induced by activated microglia via the JAK2/STAT3 signalling pathway. Microglia were cultured in oxygen-glucose deprivation (OGD) medium, which acted as a conditioning medium (CM) to activate PC12 cells, to investigate the expression of apoptosis and JAK2/STAT3 signalling-related proteins. We observed that PC12 cells apoptosis in CM was significantly increased, the expression and fluorescence intensity of the pro-apoptotic protein Bax and apoptosis-related protein cleaved caspase-3 were increased, and expression of the anti-apoptotic protein B-cell lymphoma-2 (Bcl-2) was decreased. Phosphorylation levels and fluorescence intensity of the JAK2/STAT3 signalling pathway-related proteins JAK2 and STAT3 decreased. After treatment with scutellarin, PC12 cells apoptosis as well as cleaved caspase-3 and Bax protein expression and fluorescence intensity decreased. The expression and fluorescence intensity of Bcl-2, phosphorylated JAK2, and STAT3 increased. AG490, a specific inhibitor of the JAK2/STAT3 signalling pathway, was used. Our findings suggest that AG490 attenuates the effects of scutellarin. Our study revealed that scutellarin inhibited OGD-activated microglia-mediated PC12 cells apoptosis which was regulated via the JAK2/STAT3 signalling pathway.
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Apigenina , Apoptose , Glucuronatos , Janus Quinase 2 , Microglia , Fator de Transcrição STAT3 , Transdução de Sinais , Animais , Apigenina/farmacologia , Fator de Transcrição STAT3/metabolismo , Janus Quinase 2/metabolismo , Glucuronatos/farmacologia , Células PC12 , Apoptose/efeitos dos fármacos , Microglia/efeitos dos fármacos , Microglia/metabolismo , Transdução de Sinais/efeitos dos fármacos , Ratos , Camundongos , Caspase 3/metabolismo , Glucose/metabolismo , Fármacos Neuroprotetores/farmacologia , Fosforilação/efeitos dos fármacos , Proteína X Associada a bcl-2/metabolismo , Tirfostinas/farmacologiaRESUMO
OBJECTIVE: SNRPA1, a subunit of spliceosome complex, has been implicated in diverse cancers, while its biological effect in LUAD remains elusive. Therefore, we sought to decipher the relationship between SNRPA1 expression and the prognosis of patients with LUAD and reveal the underlying molecular mechanism. MATERIALS AND METHODS: Based on the clinical data from TCGA databases, the multivariate Cox model was constructed to screen the prognostic value of SNRPA1. qRT-PCR and immunohistochemical staining were used to examine SNRPA1 mRNA and protein expression in LUAD. The effect of SNRPA1 on LUAD cell proliferation, migration, and epithelial mesenchymal transformation were examined using colony formation assays, wound healing, and western blot assays, respectively. Finally, the influence of SNRPA1 on LUAD immune microenvironment were validated from the Tumor Immune Estimation Resource database. RESULTS: SNRPA1 was significantly upregulated in both LUAD tissues and cell lines, and highly expressed SNRPA1 contributed to poor prognosis of LUAD patients. In vitro, SNRPA1 knockdown inhibited the proliferation and migration, as well as delayed the EMT differentiation of LUAD cells. Lastly, SNRPA1 was found to be positively associated with immune infiltration and some immune-check-point markers. CONCLUSIONS: Our findings indicate that SNRPA1 may be a new biomarker for prognostic prediction and a potential therapeutic target in the treatment of LUAD.
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Adenocarcinoma de Pulmão , Neoplasias Pulmonares , Humanos , Adenocarcinoma de Pulmão/genética , Western Blotting , Proliferação de Células/genética , Bases de Dados Factuais , Neoplasias Pulmonares/genética , Prognóstico , Microambiente TumoralRESUMO
Keratin 7 (KRT7) is a member of the keratin gene family. KRT7 is abnormally expressed in various types of cancer and promotes the malignant progression of tumors. However, the role of KRT7 in ovarian cancer remains unclear. The present study aimed to validate the role of KRT7 in ovarian cancer progression. KRT7 expression levels in patients with ovarian cancer were analyzed using data obtained from the Human Protein Atlas and The Cancer Genome Atlas databases. KRT7 mRNA and protein expression levels were upregulated in ovarian cancer tissue compared with normal tissue. KRT7 expression was associated with the grading, staging and poor prognosis of ovarian cancer. The differentially expressed genes affected by KRT7 were primarily enriched in the functions of cell migration, cell adhesion and cell growth. In vitro studies, including a CCK8 assay, were used to detect cell proliferation. In addition, wound healing and transwell assays were performed to analyze cell migration. The results demonstrated that KRT7 overexpression was associated with increased proliferation, migration and epithelialmesenchymal transition (EMT) of ovarian cancer cells, and the migration and EMT of ovarian cancers cells were decreased following knockdown with KRT7 small interfering RNA. In vivo, knockdown of KRT7 inhibited tumor growth of ovarian cancer. Furthermore, KRT7 regulated EMT in ovarian cancer via the TGFß/Smad2/3 pathway, and regulated cellmatrix adhesion through integrinß1focal adhesion kinase signaling. These results suggest that KRT7 may be a potential molecular marker for prognosis prediction in patients with ovarian cancer.
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Biomarcadores Tumorais/metabolismo , Queratina-7/metabolismo , Neoplasias Ovarianas/patologia , Animais , Biomarcadores Tumorais/genética , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Transição Epitelial-Mesenquimal , Feminino , Técnicas de Silenciamento de Genes , Humanos , Estimativa de Kaplan-Meier , Queratina-7/genética , Camundongos , Neoplasias Ovarianas/mortalidade , Ovário/patologia , Prognóstico , Transdução de Sinais , Proteína Smad2/metabolismo , Proteína Smad3/metabolismo , Regulação para Cima , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BACKGROUND: Circular RNA (circRNA) has emerged as an important regulator in the progression of human diseases. However, the role of circRNAs in ovarian cancer remains largely unknown. MATERIALS AND METHODS: DNA sequencing and PCR were used to identify the existence and expression of circKRT7. The targeting relationship between circKRT7/miR-29a-3p and miR-29a-3p/COL1A1 was verified by fluorescence reporter assay. In vitro, colony formation, transwell and wound healing assay were used to detect the effects of circKRT7 and miR-29a-3p on the proliferation, migration and invasion ability of ovarian cancer cells. In vivo, xenograft tumor model was performed to validate the role of circKRT7 and miR-29a-3p in tumor growth. RESULTS: We found that circKRT7 can promote the proliferation and metastasis of ovarian cancer cells by absorbing miR-29a-3p, which leads to the up-regulation of COL1A1. In vitro, knock-down of circKRT7 can inhibit the migration and invasion of ovarian cancer cells. This effect of circKRT7 is achieved by adsorbing miR-29a-3p and subsequently COL1A1 release. In vivo experiments, the reduction of circKRT7 expression can also slow tumor growth, and this inhibition was partly counteracted after miR-29a-3p repression. CONCLUSION: Overall, circKRT7 promotes EMT-related cell progression by absorbing miR-29a-3p in ovarian cancer. This suggests the crucial role of circular RNA in the malignant evolution in cancer.
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BACKGROUND: Long non-coding RNA (lncRNAs) are involved in the development and progression of numerous tumors. Nevertheless, their role in ovarian cancer (OC) needs further study. METHODS: A pivotal lncRNA that modulated OC to metastasize was determined in this research, and its potential mechanism was inquired by qRT-PCR, CCK-8, EdU, Transwell assay, wound healing assay and Western blot assay. RESULTS: In our study, the GSE119054 microarray was analyzed, and LINC00963 showed a significant higher level in ovarian cancer tissues compared with controls. So LINC00963 was selected as research object. It was discovered that LINC00963 displayed a close relationship with unfavorable prognosis, and it was prominently raised in OC tissues of patients with lymph node metastasis. What's more, LINC00963 downregulation in OC cells inhibited cell migration and invasion and inverted EMT triggered by TGF-ß1. LINC00963 downregulation also inhibited tumorigenesis in nude mice. In addition, results show that LINC00963 is a cytoplasmic lncRNA that shares the miRNA response elements (MREs) of miR-378g with CHI3L1, which is conï¬rmed by a luciferase reporter assay and AGO2-dependent RNA immunoprecipitation (RIP). CONCLUSION: On the whole, our results demonstrate an explicit oncogenic role of LINC00963 in ovarian cancer tumorigenesis via competition with miR-378g, suggesting a new regulatory mechanism of LINC00963 and providing a potential therapeutic target for ovarian cancer patients.
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RATIONALE: We report two rare cases of enchondroma protuberans originating from phalanxes. PATIENT CONCERNS: The patients visited doctors for a palpable mass in their phalanx without any pain or discomfort. DIAGNOSES: Biopsy is the gold standard for the diagnosis of enchondroma protuberans. Radiographs usually provide important imaging information, while studied on the ultrasound manifestation of enchondroma protuberans are still limited. In our cases, significant information about ultrasound manifestation of enchondroma protuberans were presented. Sonographic examination of enchondroma protuberans revealed a hypoechoic mass located in and beyond the medullary cavity of bone through the interrupted bone cortex, and blood flow signals were usually not abundant. INTERVENTIONS: Patients were subsequently referred for surgical removal and the masses were confirmed by following pathological examination. OUTCOMES: After surgery, the patients recovered well with no relapse within 2 years. LESSONS: Enchondroma protuberans is a rare form of benign enchondroma. Enchondroma protuberans can present as an intramedullary hypoechoic mass extending to the surrounding soft tissue via the discontinuous cortex line on ultrasound. Ultrasound can provide important information for the diagnosis of enchondroma protuberans.
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Neoplasias Ósseas , Doenças das Cartilagens , Condroma , Falanges dos Dedos da Mão , Adulto , Biópsia/métodos , Neoplasias Ósseas/diagnóstico por imagem , Neoplasias Ósseas/patologia , Neoplasias Ósseas/secundário , Doenças das Cartilagens/diagnóstico , Doenças das Cartilagens/patologia , Condroma/diagnóstico , Condroma/patologia , Diagnóstico Diferencial , Feminino , Falanges dos Dedos da Mão/diagnóstico por imagem , Falanges dos Dedos da Mão/patologia , Humanos , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de Neoplasias , Radiografia/métodos , Ultrassonografia/métodosRESUMO
OBJECTIVE: To compare the ultrasonic features of enthesitis between psoriatic arthritis and psoriasis vulgaris. METHODS: A total of 39 patients with psoriatic arthritis (PsA group), 60 with psoriasis vulgaris (non-PsA group) and 60 healthy people (control group) participated in this study. They were examined by two-dimensional and color Doppler ultrasound on the entheses of bilateral femoral quadriceps tendons, patella tendons, Achilles tendons, plantar fasciae, common flexor tendons and common extensor tendons. RESULTS: About 45% (27 cases) healthy controls had enthesitis, with Achilles tendons and femoral quadriceps tendons being most likely affected. No blood flow signal was observed on the affected sites. About 63% (38 cases) of non-PsA patients had enthesitis, with Achilles tendons and femoral quadriceps tendons being most likely affected. Blood flow signals were observed on 4 affected sites. More than 84% (33 cases) PsA patients had enthesitis, with all locations being likely affected but mostly on Achilles tendons, femoral quadriceps tendons, and plantar fasciae. Blood flow signals were observed on 18 affected sites. The differences in prevalence of enthesitis were statistically significant (PsA group>non-PsA group>control group, all P<0.01), although the differences in tendon hypoechogenicity and enthesophytes among the groups showed no statistical significance. PsA and non-PsA patients were more likely to have tendon thickening than the controls (both P<0.01); but no difference appeared between PsA and non-PsA patients. PsA patients had higher prevalence of intratendinous calcifications, bony erosions and color Doppler signals than non-PsA patients and the controls (all P<0.01). CONCLUSION: Enthesitis in healthy people and non-PsA patients are most likely to affect Achilles tendon and femoral quadriceps tendons. By contrast, Achilles tendons, femoral quadriceps tendon and plantar fascia are more likely to be affected in patients with PsA. PsA patients have high prevalence of enthesitis and are more likely to have intratendinous calcifications, bony erosions and color Doppler signals.
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Artrite Psoriásica/diagnóstico por imagem , Psoríase/diagnóstico por imagem , Tendão do Calcâneo/diagnóstico por imagem , Tendão do Calcâneo/patologia , Estudos de Casos e Controles , Humanos , Músculo Esquelético/diagnóstico por imagem , Músculo Esquelético/patologia , UltrassonografiaRESUMO
OBJECTIVES: To explore the value of high frequency color doppler ultrasonography in differentiating benign and malignant skin solid tumors. METHODS: Clinical and ultrasonic data of cutaneous solid tumors confirmed by pathology in our hospital were collected. The differences in clinical and sonographic features between benign and malignant tumors were statistically analyzed. RESULTS: A total of 512 patients, involving 527 cases of skin solid tumors, were enrolled in this study. The ultrasonic detected 99.43% of the cases, with 99.02% accuracy in locating the lesions. The benign and malignant tumors showed differences in patient age, location, multiple occurance, location and depth, surface skin condition, tumor size, echo, morphology, uniformity, calcification, blood flow status, tumor rear area and peripheral echo, and pathological requests ( P<0.05). CONCLUSIONS: High frequency ultrasound has excellent detection rate of skin tumors, which can locate invasion depth of skin accurately. Benign and malignant skin tumors show differences in a number of clinical and ultrasound features.
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Neoplasias Cutâneas/diagnóstico por imagem , Ultrassonografia Doppler em Cores , Diagnóstico Diferencial , Humanos , Sensibilidade e Especificidade , Pele/patologiaRESUMO
BACKGROUND: We aimed to investigate the function of miR-1307 in chemoresistance and to explore its chemoresistance mechanism in ovarian cancer. METHODS: IC50 determination was used to test the chemoresistance profling in ovarian cancer cells. QRT-PCR or western blot was used to validate the expression level of miR-1307 and candidate gene or protein. Colony formation assay and FITC-labeled enhanced Annexin V immunofluorescence were used to compare cell proliferation and apoptosis ability, respectively. The potential target gene and its biological function of miRNA-1307 were also analyzed. Bioinformatics and Luciferase Reporter Gene Assay were conducted to validate the regulation of miRNA-1307 on the ING5 expression. Xenografts assay was used to demonstrate the inhibiting effect of miR-1307 ASO and Taxol therapy against ovarian cancer in vivo. RESULTS: MiR-1307 was over-expressed in chemoresistant ovarian cancer cell line A2780/Taxol, and over-expression or loss of miR-1307 promoted or inhabited chemoresistance. And we also found that the over-expression of miR-1307 promoted proliferation and inhibited apoptosis in ovarian cancer cells. Besides, we demonstrated that ING5 was a direct target of miR-1307 and miR-1307 down-regulated the ING5 expression in ovarian cancer cells. Additionally, we showed that ING5 inhibited cell proliferation, promoted cell apoptosis and inhabited chemoresistance reversely. Furthermore, the up-regulated ability of cell apoptosis and down-regulated ability of chemoresistance following the loss of miR-1307 was reversed by adding ING5 siRNA in vitro. Finally, we proved the inhibiting effect of miR-1307 ASO and Taxol therapy by increasing the ING5 expression against ovarian cancer through xenografts assay in vivo. CONCLUSION: Our results suggested that miR-1307 could promote ovarian cancer chemoresistance by targeting the ING5 expression and miR-1307 might serve as a therapeutic target for ovarian cancer.
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Resistencia a Medicamentos Antineoplásicos/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Neoplasias Ovarianas/genética , Interferência de RNA , Fatores de Transcrição/genética , Proteínas Supressoras de Tumor/genética , Regiões 3' não Traduzidas , Animais , Antineoplásicos/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Apoptose/genética , Sítios de Ligação , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular/genética , Modelos Animais de Doenças , Feminino , Expressão Gênica , Humanos , Camundongos , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/patologia , Paclitaxel/farmacologia , Transfecção , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Neural stem/progenitor cell (NSC) transplantation has been shown to effectively improve neurological function in rats with hypoxic-ischemic brain damage. Vascular endothelial growth factor (VEGF) is a signaling protein that stimulates angiogenesis and improves neural regeneration. We hypothesized that transplantation of VEGF-transfected NSCs would alleviate hypoxic-ischemic brain damage in neonatal rats. We produced and transfected a recombinant lentiviral vector containing the VEGF165 gene into cultured NSCs. The transfected NSCs were transplanted into the left sensorimotor cortex of rats 3 days after hypoxic-ischemic brain damage. Compared with the NSCs group, VEGF mRNA and protein expression levels were increased in the transgene NSCs group, and learning and memory abilities were significantly improved at 30 days. Furthermore, histopathological changes were alleviated in these animals. Our findings indicate that transplantation of VEGF-transfected NSCs may facilitate the recovery of neurological function, and that its therapeutic effectiveness is better than that of unmodified NSCs.
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A 31-month-old boy with Wilms' tumor (WT), which primarily had originated from the left kidney, was treated with nephrectomy and adjuvant chemotherapy. 2 months after nephrectomy, a left scrotal mass was found at routine follow-up. High-frequency sonography examination revealed an enlarged left testis with a heterogeneous texture and a hypoechoic solid mass in the left scrotum. Moreover, hypervascular signals presented in both the left testis and the mass on color Doppler flow imaging. Left orchiectomy was performed for suspected intrascrotal metastasis of WT, which was confirmed by histopathology examination. This was the first case of intrascrotal metastasis of WT reported in China with a detailed ultrasound description. Meanwhile, this study also reviewed the comparable diagnostic methods of intrascrotal metastasis of WT found in the English literature.
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Neoplasias dos Genitais Masculinos/diagnóstico por imagem , Neoplasias dos Genitais Masculinos/secundário , Neoplasias Renais/diagnóstico por imagem , Neoplasias Renais/patologia , Escroto , Tumor de Wilms/diagnóstico por imagem , Tumor de Wilms/secundário , Humanos , Lactente , Masculino , UltrassonografiaRESUMO
OBJECTIVE: To investigate the effect of intraventricular injection of human dental pulp stem cells (DPSCs) on hypoxic-ischemic brain damage (HIBD) in neonatal rats. METHODS: Thirty-six neonatal rats (postnatal day 7) were assigned to control, HIBD, or HIBD+DPSC groups (nâ=â12 each group). For induction of HIBD, rats underwent left carotid artery ligation and were exposed to 8% to 10% oxygen for 2 h. Hoechst 33324-labeled human DPSCs were injected into the left lateral ventricle 3 days after HIBD. Behavioral assays were performed to assess hypoxic-ischemic encephalopathy (HIE), and on postnatal day 45, DPSC survival was assessed and expression of neural and glial markers was evaluated by immunohistochemistry and Western blot. RESULTS: The HIBD group showed significant deficiencies compared to control on T-maze, radial water maze, and postural reflex tests, and the HIBD+DPSC group showed significant improvement on all behavioral tests. On postnatal day 45, Hoechst 33324-labeled DPSC nuclei were visible in the injected region and left cortex. Subsets of DPSCs showed immunostaining for neuronal (neuron-specific enolase [NSE], Nestin) and glial markers (glial fibrillary acidic protein [GFAP], O4). Significantly decreased staining/expression for NSE, GFAP, and O4 was found in the HBID group compared to control, and this was significantly increased in the HBID+DPSC group. CONCLUSION: Intraventricular injection of human DPSCs improves HIBD in neonatal rats.
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Células-Tronco Adultas/transplante , Polpa Dentária/citologia , Hipóxia-Isquemia Encefálica/terapia , Adolescente , Adulto , Células-Tronco Adultas/fisiologia , Animais , Animais Recém-Nascidos , Antígenos de Diferenciação/metabolismo , Diferenciação Celular , Proteína Glial Fibrilar Ácida/metabolismo , Humanos , Hipóxia-Isquemia Encefálica/fisiopatologia , Hipóxia-Isquemia Encefálica/psicologia , Injeções Intraventriculares , Aprendizagem em Labirinto , Nestina/metabolismo , Ratos , Ratos Sprague-Dawley , Reflexo Anormal , Adulto JovemRESUMO
BACKGROUND: There is a large number (1.5 million per year) of premature births in China. It is necessary to obtain the authentic incidences of intraventricular hemorrhage (IVH) and periventricular leukomalacia (PVL), the common brain injuries, in Chinese premature infants. The present multicenter study aimed to investigate the incidence of brain injuries in premature infants in ten urban hospitals in China. METHODS: The research proposal was designed by the Subspecialty Group of Neonatology of Pediatric Society of the Chinese Medical Association. Ten large-scale urban hospitals voluntarily joined the multicenter investigation. All premature infants with a gestational age ≤ 34 weeks in the ten hospitals were subjected to routine cranial ultrasound within three days after birth, and then to repeated ultrasound every 3-7 days till their discharge from the hospital from January 2005 to August 2006. A uniform data collection sheet was designed to record cases of brain injuries. RESULTS: The incidences of overall IVH and severe IVH were 19.7% (305/1551) and 4.6% (72/1551), respectively with 18.4% (56/305) for grade 1, 58.0% (177/305) for grade 2, 17.7% (54/305) for grade 3 and 5.9% (18/305) for grade 4 in nine hospitals. The incidences of overall PVL and cystic PVL were 5.0% (89/1792) and 0.8% (14/1792) respectively, with 84.3% (75/89) for grade 1, 13.5% (12/89) for grade 2, and 2.2% (2/89) for grade 3 in the ten hospitals. The statistically significant risk factors that might aggravate the severity of IVH were vaginal delivery (OR=1.883, 95% CI: 1.099-3.228, P=0.020) and mechanical ventilation (OR=4.150, 95% CI: 2.384-7.223, P=0.000). The risk factors that might result in the development of cystic PVL was vaginal delivery (OR=21.094, 95% CI: 2.650-167.895, P=0.000). CONCLUSIONS: The investigative report can basically reflect the incidence of brain injuries in premature infants in major big cities of China. Since more than 60% of the Chinese population live in the rural areas of China, it is expected to undertake a further multicenter investigation covering the rural areas in the future.
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Lesões Encefálicas/epidemiologia , China , Feminino , Idade Gestacional , Hospitais Urbanos , Humanos , Incidência , Recém-Nascido , Recém-Nascido Prematuro , MasculinoRESUMO
Cerebral palsy (CP) is a very common neural system development disorder that can cause physical disability in human. Here, we studied the neuroprotective effect of vascular endothelial growth factor (VEGF)-transfected neural stem cells (NSCs) in newborn rats with cerebral palsy (CP). Seven-day-old Sprague-Dawley rats were randomly divided into four groups: sham operation (control group), PBS transplantation (PBS group), VEGF+NSCs transplantation (transgene NSCs group) and NSCs transplantation groups (NSCs group). PBS, Transgene NSCs and NSCs groups respectively received stereotactic injections of PBS, lentiviral vector (pGC-FU-VEGF) infected NSCs or a NSCs suspension in the left sensory-motor cortex 3 days after CP model was established. The NSCs activity, their impacts on neural cell growth and apoptosis, brain development and animal behaviors were examined on the animals up to age 35-days. As expected, unilateral carotid artery occlusion plus hypoxia (cerebral palsy model) resulted in severe neural developmental disorders, including slowed growth, increased in cortical neuron apoptosis, decreased cerebral cortex micro-vessel density and retarded behavior developments. Transplantation of NSCs not only resulted in increases in VEGF protein expression in rat brains, but also largely prevented the behavioral defects and brain tissue pathology that resulted from cerebral palsy procedure, with animals received VEGF transfected NSCs always being marginally better than these received un-transfected cells. In conclusion, NSCs transplantation can partially prevent/slow down the brain damages that are associated with CP in the newborn rats, suggesting a new possible strategy for CP treatment.
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Paralisia Cerebral/prevenção & controle , Fármacos Neuroprotetores/uso terapêutico , Transplante de Células-Tronco/métodos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Animais Recém-Nascidos , Encéfalo/metabolismo , Encéfalo/patologia , Doenças das Artérias Carótidas/complicações , Paralisia Cerebral/etiologia , Paralisia Cerebral/metabolismo , Paralisia Cerebral/patologia , Modelos Animais de Doenças , Feminino , Vetores Genéticos/fisiologia , Hipóxia/complicações , Marcação In Situ das Extremidades Cortadas , Masculino , Transtornos Mentais/etiologia , Microvasos/patologia , Neovascularização Patológica/etiologia , Gravidez , Ratos , Ratos Sprague-Dawley , Fatores de Tempo , Transfecção , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
Previous studies have showed that vascular endothelial growth factor (VEGF) displayed neurotrophic and neuroprotective activities. To examine whether target delivery of VEGF gene directly into brain may prevent ischemic brain damage, the VEGF expression adenoviral vectors, AVHP.VEGF-with 476bp of the human preproendothelin-1 (ppET-1) promoter and 35bp of the hypoxia-reponse element (HRE) driving VEGF expression and CMV.VEGF were transferred into hypoxic-induced ischemic (HI) rat brains. Seven-day-old rats that were underwent left carotid ligation followed by 2h of hypoxic stress (8% O(2) at 37 degrees C) were received VEGF adenoviral vectors or buffer (PBS) injection 3 days after HI. The body weight, VEGF expression, neuronal apoptosis, cerebral morphology and brain functional assays were performed between 7 and 28 days after HI. There were remarkable increases in the body weight and VEGF protein expression, and decrease in the number of TUNEL-positive cells in the VEGF vector groups as compared with PBS group. The VEGF vector groups also had better brain functional performs than PBS group. The better performs by the animals that received VEGF vectors may be directly linked to the inhibitory effect of VEGF on neuronal apoptosis because the animals had less neural loss in the cortex and hippocampal CA1 region as compared with PBS group. Overall, these results indicated that over-expression of VEGF in the brain exerted a neuroprotective effect and promoted neural functional recovery in neonatal rats after hypoxic-ischemic brain damage, suggesting that in vivo target VEGF gene transfer to brain may be a promising approch for the treatment of such implications.
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Adenoviridae/genética , Vetores Genéticos , Hipóxia-Isquemia Encefálica/terapia , Transdução Genética/métodos , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Animais Recém-Nascidos , Apoptose/genética , Apoptose/fisiologia , Peso Corporal/genética , Peso Corporal/fisiologia , Encéfalo/patologia , Encéfalo/fisiopatologia , Endotelina-1/genética , Humanos , Hipóxia-Isquemia Encefálica/patologia , Hipóxia-Isquemia Encefálica/fisiopatologia , Neurônios/fisiologia , Regiões Promotoras Genéticas , Ratos , Ratos Sprague-Dawley , Recuperação de Função Fisiológica/genética , Recuperação de Função Fisiológica/fisiologia , Fatores de Tempo , Resultado do TratamentoRESUMO
OBJECTIVE: To investigate the therapeutical effect of adenovirus-mediated vascular endothelial growth factor (VEGF)165 gene transplantation in treatment of hypoxic-ischemic brain damage. METHODS: Recombinant vector of adenovirus-mediated VEGF165 gene (Ad-VEGF) was constructed by bacterial homologous recombination technology. Sixty 7-day-old Sprague-Dawley rats were randomly divided into 3 equal groups: hypoxic-ischemic brain damage (HIBD) group undergoing ligation of the left common carotid artery and inhalation of 8% oxygen for 2 hours, Ad-VEGF group undergoing injection of Ad-VEGF into the left sensorimotor cortex with the help of stereo-positioner 3 days after hypoxia-ischemia (HI), and sham operation group. Seven days after transplantation, 5 rats from each group were killed with their left brains taken out. The VEGF protein expression was detected by immunohistochemistry, and the neuron apoptosis was detected by terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nickel end labeling (TUNEL). Since the age of 28 days T-maze foraging test was conducted. At the age of 34 days, sensorimotor tests were performed. After the behavioral tests all the rats were killed. The number of neurons in the CA1 region of the hippocampus and cerebral cortex was detected by Nissl's staining. RESULTS: Immunohistochemistry showed that the density levels of VEGF positive cells in cerebral cortex and hippocampus of the Ad-VEGF group were (68.09 +/- 3.37) and (68.37 +/- 3.17) respectively, both significantly higher than those of the HIBD group [(24.65 +/- 3.14) and (25.14 +/- 1.86) respectively, both P < 0.05]. TUNEL showed that the number of apoptotic neurons of the Ad-VEGF group was (151.4 +/- 21.7), significantly lower than that of the HIBD group [(264.4 +/- 16.3), P < 0.05]. Behavioral tests showed the percentages of accuracy on day 4 of the Ad-VEGF and sham operation groups were both significantly higher than that of the HIBD group (both P < 0.01). Nissl's staining showed that the numbers of neurons per unit area in the hippocampal CA1 area and cortex of the Ad-VEGF group were (70.6 +/- 2.3) and (95.1 +/- 2.8) respectively, both significantly higher than those of the HI group [(55.3 +/- 2.1) and (70.1 +/- 2.7) respectively, both P < 0.05]. CONCLUSION: Adenovirus vector-mediated VEGF gene therapy increases the VEGF protein expression, decreases neuron apoptosis, improves the long-term behavioral function after brain damage, and reduces hypoxic ischemic brain injury, thus possessing neuroprotective effects.
Assuntos
Terapia Genética , Hipóxia-Isquemia Encefálica/terapia , Fator A de Crescimento do Endotélio Vascular/genética , Adenoviridae/genética , Animais , Feminino , Hipóxia-Isquemia Encefálica/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
OBJECTIVE: To investigate the protective effects of adenovirus-mediated vascular endothelial growth factor (Ad-VEGF)165 gene transfer against hypoxic-ischemic brain damage (HIBD) in neonatal rats. METHODS: Ad-VEGF recombinant adenovirus was constructed by bacterial homologous recombination technology. Seven-day-old Sprague-Dawley rats were randomly assigned to 4 groups: sham-operated (n=20), HIBD (n=25), buffer-treated (n=20), and Ad-VEGF-treated (n=25). The HIBD model was prepared by permanent occlusion of left common carotid artery, followed by exposure to 8% oxygen for 2 hrs. In the Ad-VEGF-treated and the Buffer-treated groups, 2 microL recombinant adenovirus suspension or buffer was injected into the left sensorimotor cortex of the rat brain 3 days after HIBD. Seven days after transplantation, VEGF165 mRNA expression was detected using RT-PCR. Neuronal apoptosis was detected by the terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nickel end labeling (TUNEL). CD34 and VEGF protein were detected using immunohistochemistry. Microvascular density in the cerebral cortex was measured based on CD34 positive cells. A radial arm maze test was performed from 30 postnatal days to evaluate long-term learning and memory functions. At 35 postnatal days, the rats were sacrificed for cerebral histological examinations by hematoxylin and eosin. RESULTS: The expression of VEGF165 mRNA increased in the Ad-VEGF-treated group more than in the untreated HIBD and the buffer-treated groups (p<0.05). The number of apoptotic neurons was less in the Ad-VEGF-treated group compared with that in the untreated HIBD and the buffer-treated groups (p<0.05). Microvascular density and VEGF positive cells increased in the Ad-VEGF-treated group compared with that in the untreated HIBD and the buffer-treated groups (p<0.05). In the radial arm maze test, the Ad-VEGF-treated group had more improved achievements than the HIBD and the buffer groups (p<0.05). Neuronal degeneration and necrosis were lessened in the Ad-VEGF-treated group compared with the HIBD and the buffer groups. CONCLUSIONS: Ad-VEGF gene transfer can increase the expression of VEGF mRNA and VEGF protein, decrease neuronal apoptosis, and increase angiopoiesis in the brain. This attenuates brain damage and improves long-term learning and memory functions in neonatal rats after HIBD.
Assuntos
Adenoviridae/genética , Terapia Genética , Hipóxia-Isquemia Encefálica/terapia , Fator A de Crescimento do Endotélio Vascular/genética , Animais , Animais Recém-Nascidos , Córtex Cerebral/irrigação sanguínea , Córtex Cerebral/química , Feminino , Marcação In Situ das Extremidades Cortadas , Masculino , Fármacos Neuroprotetores , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Fator A de Crescimento do Endotélio Vascular/análiseRESUMO
OBJECTIVE: To study the effect of intracerebral transplantation of bone marrow stromal cells (BMSCs) on brain white matter of neonatal rats with hypoxic-ischemic brain damage (HIBD). METHODS: Thirty-four 7-day-old neonatal rats were randomly assigned to three groups: normal control (n=10), HIBD (n=12) and HIBD+BMSCs transplantation (n=12). The HIBD and the HIBD+BMSCs transplantation group rats were subjected to left carotid artery ligation, followed by hypoxia exposure for 2 hrs, in order to induce HIBD. The rats in the HIBD+BMSCs transplantation group received transplantation of BMSCs labeled nucleus with Hochest 33324 into the left hippocampus 24 hrs after HIBD induction. Myelin basic protein (MBP) expression in the left corpus callosum and the subcortical white matter and the number of oligodendrocyte precursors positively stained O4 in the left periventricular area and the subcortical white matter were detected by immunohistochemistry at ages of 45 days. RESULTS: The labeled BMSCs survived and were found mainly in the left hemisphere 37 days after transplantation. The positive rate of O4 expressed by the transplanted BMSCs was 3.70+/-1.09%. More hypomyelination in the left corpus callosum and the subcortical white matter, and less number of O4 positive oligodendrocytes in the left periventricular area and the subcortical white matter were found in the HIBD group compared with the normal control group (P<0.01). The HIBD rats receiving BMSCs transplantation had increased O4 positive oligodendrocytes in the left periventricular area and the subcortical white matter and improved MBP immunoreactivity in the left corpus callosum and the subcortical white matter compared with the HIBD group (P<0.01). CONCLUSIONS: Intracerebral transplantation of BMSCs can improve brain white matter damage in neonatal rats with HIBD.
Assuntos
Células da Medula Óssea/fisiologia , Encéfalo/patologia , Hipóxia-Isquemia Encefálica/terapia , Células Estromais/transplante , Animais , Animais Recém-Nascidos , Antígenos de Diferenciação/análise , Hipóxia-Isquemia Encefálica/metabolismo , Hipóxia-Isquemia Encefálica/patologia , Imuno-Histoquímica , Proteína Básica da Mielina/análise , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To study the effects of heat shock preconditioning on the expression of heat shock protein-70 (HSP70) and apoptosis of the neuron in experimental autoimmune encephalomyelitis (EAE) rats. METHODS: Thirty-six Wistar rats were randomly divided into control, EAE and heat shock preconditioning groups (n=12 each). The EAE animal model was induced with guinea pig myelin basic protein. Heat shock preconditioning was performed 24 hrs prior to the EAE model inducement. No treatment was done in the control group. The neurological signs were observed after immunization. The spinal cords were removed and stained with hematoxylin and eosin. HSP70 was detected by immunohistochemistry. Apoptosis of the neuron was measured by TUNEL. RESULTS: Heat shock preconditioning significantly alleviated clinical signs and neuronal injury. HSP70 expression in the heat shock preconditioning group was significantly higher than in the untreated EAE group (21.08 +/- 0.87 vs 10.17 +/- 0.51; P < 0.01). Heat shock preconditioning suppressed apoptosis of the neuron compared with the EAE group (apoptosis rate: 21.92 +/- 1.00% vs 58.92 +/- 1.67%; P < 0.01). CONCLUSIONS: Heat shock preconditioning might improve the neurological outcome in EAE rats, possibly through the induction of HSP70 synthesis and the reduction of apoptosis of the neuron in spinal cords.