RESUMO
Bone mesenchymal stem cells (MSCs) are an excellent donor graft source due to their potential for selfrenewal and multidirectional differentiation. However, it is difficult to obtain high quality MSCs and to induce them to differentiate into neuronlike cells. Fasudil, a Rho kinase inhibitor, exhibits therapeutic potential in spinal cord injuries and stroke. The present study investigated the effect of fasudil on the differentiation of MSCs into neuronlike cells. MSCs were obtained from rat femur marrow, expanded in culture medium, and used at the third passage for subsequent experiments. MSCs were preinduced with 10 ng/ml basic fibroblast growth factor (bFGF) for 24 h, which was followed by induction with fasudil. A control untreated group and a group treated with fasudil + XAV939, a Wnt/ßcatenin pathway inhibitor, were also used in the present study. Reverse transcriptionquantitative polymerase chain reaction (RTqPCR), western blot analysis and immunofluorescence staining were performed in order to detect neuronspecific markers, including neuronspecific enolase (NSE), nestin and neurofilamentM (NFM). Following induction with fasudil, neuronlike cell morphology was observed. In the fasudil + XAV939 and control groups, no obvious changes in cell shape were observed. The results of RTqPCR, western blot analysis and immunofluorescence staining indicated that expression of the neuronspecific markers NSE, nestin and NFM was detected in the fasudil group. The differentiation of MSCs into neuronlike cells induced by fasudil was eliminated when the Wnt/ßcatenin pathway was inhibited. The present study demonstrated that fasudil may induce MSCs to differentiate into neuronlike cells, however further studies are required to determine the specific mechanisms involved in the effect of fasudil on the Wnt/ßcatenin pathway. In addition, further research is required to examine the functional characteristics of the induced neuronlike cells, in order to establish their suitability for clinical treatments in the future.
Assuntos
Diferenciação Celular , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Via de Sinalização Wnt , Animais , Biomarcadores , Ciclo Celular , Imunofluorescência , Imunofenotipagem , Masculino , Microscopia , RatosRESUMO
BACKGROUND: Cytochrome P450 (CYP) 2C19 is a very important drug metabolizing enzyme. Although the single nucleotide polymorphisms (SNPs) of CYP2C19 G681A and G636A have been suggested that they may increase the incidence of cardiovascular events, the relationship between SNPs in CYP2C19 and cerebral ischemic stroke (CIS) are unclear. The aim of this study was to investigate the correlation between the distribution of G681A and G636A polymorphisms in CYP2C19 gene and the risk of CIS in Chinese. METHODS: The peripheral blood DNA was extracted from 299 patients with CIS and 295 healthy controls. The genotyping was conducted using the polymerase chain reaction-restriction fragment length polymorphism. The sampled sequencing was applied to verify the correctness of genotyping results. Both the genotype and allele distributions were compared in patients with CIS and healthy controls. RESULTS: The frequencies of CYP2C19 681AA (11.7% vs. 2.7%; P = 0.000), 636AA (4.0% vs. 0.7%; P = 0.007), 636AG (7.0% vs. 2.2%; P = 0.038) genotype, CYP2C19 681A (30.9% vs. 20.8%; P = 0.000) and 636A (13.0% vs. 5.8%; P = 0.000) allele in the CIS group are significantly higher than those in the controls. The frequencies of CYP2C19 681AA (16.7% vs. 8.6%; P = 0.036), CYP2C19 636AA (7.0% vs. 2.2%; P = 0.038) genotype, CYP2C19 681A (36.4% vs. 27.6%; P = 0.023) and CYP2C19 636A (17.5% vs.10.3%; P = 0.010) allele in the recurrent stroke group are significantly higher than those in the first onset group. Multivariate logistic regression analysis of risk factors for cerebral ischemic stroke and recurrent stroke respectively suggests that the CYP2C19 681AA genotype may be an independent risk factor for CIS (OR = 6.179, 95% CI: 2.285 ~ 16.708; P = 0.000) and recurrent stroke (OR = 2.305, 95% CI: 1.121 ~ 4.743; P = 0.023). CONCLUSIONS: The AA genotype and A allele of CYP2C19 G681A may be related to the occurrence and recurrence of cerebral ischemic stroke.
Assuntos
Hidrocarboneto de Aril Hidroxilases/genética , Isquemia Encefálica/genética , Polimorfismo de Nucleotídeo Único , Acidente Vascular Cerebral/genética , Idoso , Povo Asiático/genética , Estudos de Casos e Controles , Citocromo P-450 CYP2C19 , Feminino , Frequência do Gene , Estudos de Associação Genética , Predisposição Genética para Doença , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Recidiva , Fatores de Risco , Análise de Sequência de DNARESUMO
Increased levels of asymmetric dimethylarginine (ADMA) have been observed in patients with acute ischemic stroke. We aimed to investigate the correlation between ADMA and ischemic stroke, and evaluate the effect of supplementation of folic acid and vitamin B12 on concentrations of ADMA. Patients were randomized into intervention and non-intervention groups within 3 days after symptom onset. Intervention group patients were treated with folic acid (5mg daily) and vitamin B12 (500 µg twice daily) for 12 weeks. ADMA and homocysteine (Hcy) concentrations were measured before treatment (baseline) and 2 and 12 weeks after treatment. The laboratory measures were also collected from healthy controls. Eighty five subjects were enrolled in this study, from whom 72 with complete baseline and follow-up laboratory data were included in the present analysis. Thirty four patients were assigned to the intervention group and 38 patients to the non-intervention group. Sixty people were enrolled as healthy controls. Levels of ADMA and Hcy were raised (p<0.05) in patients with acute ischemic stroke. With supplementation of both folic acid and vitamin B12, the levels of ADMA and Hcy decreased significantly at 2 and 12 weeks (p<0.05). The present study reconfirmed that ADMA can be regarded as a risk biomarker for acute ischemic stroke. We observed that with supplementation of folic acid and vitamin B12, levels of ADMA were decreased in patients with acute ischemic stroke.
Assuntos
Arginina/análogos & derivados , Isquemia Encefálica/sangue , Isquemia Encefálica/tratamento farmacológico , Ácido Fólico/administração & dosagem , Acidente Vascular Cerebral/sangue , Acidente Vascular Cerebral/tratamento farmacológico , Vitamina B 12/administração & dosagem , Idoso , Arginina/sangue , Análise Química do Sangue , Suplementos Nutricionais , Feminino , Seguimentos , Homocisteína/sangue , Humanos , Masculino , Óxido Nítrico/sangue , Óxido Nítrico Sintase Tipo III/sangue , Fatores de Tempo , Resultado do TratamentoRESUMO
Kynurenic acid (KYNA), an antagonist of the α7 nicotinic acetylcholine receptor and the N-methyl-D-aspartate receptor, and 3-hydroxykynurenine (3-HK), a generator of reactive oxygen species, are neuroactive metabolites of the kynurenine pathway of tryptophan degradation. In the mammalian brain as elsewhere, both compounds derive from a common bioprecursor, L-kynurenine (L-KYN). Recent studies in rats demonstrated that D-kynurenine (D-KYN), a metabolite of the bacterial amino acid D-tryptophan, can also function as a bioprecursor of brain KYNA. We now investigated the conversion of systemically administered D-KYN to KYNA in mice and also explored the possible production of 3-HK in the same animals. Thirty min after an injection of D-KYN or L-KYN (30 mg/kg, i.p.), newly produced KYNA and 3-HK were recovered from plasma, liver, forebrain and cerebellum in all cases. Using a new chiral separation method, 3-HK produced from D-KYN was positively identified as D-3-HK. L-KYN was the more effective precursor of KYNA in all tissues and also exceeded D-KYN as a precursor of brain 3-HK. In contrast, D-KYN was more potent as a precursor of 3-HK in the liver. The production of both KYNA and 3-HK from D-KYN was rapid in all tissues, peaking at 15-30 min following a systemic injection of D-KYN. These results show that biosynthetic routes other than those classically ascribed to L-KYN can account for the synthesis of both KYNA and 3-HK in vivo. This new insight may be of significant physiological or pathological relevance.
Assuntos
Ácido Cinurênico/metabolismo , Cinurenina/análogos & derivados , Cinurenina/metabolismo , Animais , Química Encefálica/fisiologia , Catálise , Feminino , Ácido Cinurênico/sangue , Ácido Cinurênico/síntese química , Cinurenina/biossíntese , Cinurenina/sangue , Cinurenina/química , Cinurenina/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos , Estereoisomerismo , Triptofano/administração & dosagemAssuntos
Ataque Isquêmico Transitório/prevenção & controle , Prevenção Secundária/normas , Acidente Vascular Cerebral/prevenção & controle , China/epidemiologia , Humanos , Ataque Isquêmico Transitório/diagnóstico , Ataque Isquêmico Transitório/epidemiologia , Acidente Vascular Cerebral/diagnóstico , Acidente Vascular Cerebral/epidemiologiaRESUMO
OBJECTIVE: To investigate the changes in stress hormones in neurogenic pulmonary edema (NPE) and explore the clinical value of mild hypothermia therapy for treatment of NPE. METHODS: Fifty-two patients with cerebral hemorrhage patients and concomitant NPE were randomly divided into two groups for local mild hypothermia therapy (23 cases, LMH group) or routine treatment (29 cases, RT group). In the former group, local mild hypothermia therapy was applied in addition to the routine treatment. The changes of serum corticotrophin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), corticosteroid (Cor), arginine vasopressin (AVP) and blood sugar were observed before and 7 days after the treatment, and compared with those of 58 NPE-free patients with cerebral hemorrhage and 40 healthy individuals. RESULTS: Serum CRH, ACTH, Cor, and AVP levels and blood sugar in NPE patients and the NPE-free patients were all significantly higher than those in the healthy individuals (P<0.01), and the levels were significantly higher in NPE patients than in the NPE-free patients (P<0.05). In the NPE patients, the mortality rate and NIHSS score were significantly lower in RT group (P<0.01); after 7 days of treatment, both LMH and RT groups showed significant reduction in serum CRH, ACTH, Cor, and AVP levels (P<0.05), and the reduction was more conspicuous in LMH group (P<0.05). CONCLUSION: The occurrence of NPE is closely associated with stress reactions, which might be the basis of NPE. Local mild hypothermia therapy improves of the quality of life of NPE patients and also decreases the mortality of NPE possibly by inhibiting the secretion of stress hormones and stabilizing the hypothalamic-pituitary-adrenal axis.
Assuntos
Hipotermia Induzida/métodos , Hemorragias Intracranianas/terapia , Edema Pulmonar/terapia , Hormônio Adrenocorticotrópico/sangue , Adulto , Idoso , Arginina Vasopressina/sangue , Hormônio Liberador da Corticotropina/sangue , Feminino , Cabeça , Humanos , Hemorragias Intracranianas/sangue , Hemorragias Intracranianas/complicações , Masculino , Pessoa de Meia-Idade , Edema Pulmonar/sangue , Edema Pulmonar/etiologia , Resultado do TratamentoRESUMO
OBJECTIVE: To investigate the effects of GM1 on inducing adult rat bone marrow stromal cells (MSCs) to form neural progenitor cells and their differentiation. METHODS: Purified MSCs were induced by different components of basic fibroblast growth factor (bFGF) alone, GM1 alone or combination of bFGF with GM1. After 3 days' incubation, fibronectin and collagen I were detected with immunocytochemistry, and nestin was detected with immunofluorescence. Neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP) and galactose cerebroside (GalC) were detected with immunocytochemistry after 7 days' incubation. RESULTS: After induction with bFGF alone or combination of bFGF and GM1, some MSCs exhibited the phenotypes of neural progenitor cells, and then neurons and astrocytes. In these two groups, the positive cells for fibronectin and collagen I decreased markedly after 3 days' induction. At the same time, the positive cells for nestin increased markedly. After 7 days' induction, NSE and GFAP-positive cells increased significantly. Furthermore, the addition of bFGF and GM1 caused the maximal variation. However, addition of GM1 alone had no inductive effects. CONCLUSIONS: Combination of bFGF with GM1 may synergistically promote the transformation of MSCs and differentiation into neurons and astrocyte-like cells. The results suggest a promising route for the application of MSCs.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Fator 2 de Crescimento de Fibroblastos/farmacologia , Gangliosídeo G(M1)/farmacologia , Células-Tronco/fisiologia , Células Estromais/efeitos dos fármacos , Análise de Variância , Animais , Células da Medula Óssea , Diferenciação Celular/fisiologia , Células Cultivadas , Sinergismo Farmacológico , Imunofluorescência , Imuno-Histoquímica , Probabilidade , Ratos , Ratos Wistar , Sensibilidade e Especificidade , Células-Tronco/patologia , Células Estromais/fisiologia , Células Estromais/ultraestruturaRESUMO
OBJECTIVE: To evaluate the clinical efficacy and safety of huperzine Alpha in treatment of patients with mild to moderate Alzheimer disease (AD). METHODS: Two hundred and two patients with the diagnosis of possible or probable AD from 15 centers the nationwide were randomly divided into two groups: huperzine Alpha group (n = 100, given huperzine Alpha 400 micro g/day for 12 weeks) and placebo group (n = 102 ). Different scales were used to evaluate the cognitive function, activity of daily life (ADL), non-cognitive disorders, and overall clinical efficacy. Safety evaluation was conducted every 6 weeks. RESULTS: In comparison with the baseline data, there was an improvement of 4.6 points in cognition assessed by ADAS-Cog (P = 0.000); an improvement of 2.7 points by MMSE (P = 0.000), an improvement of 1.5 points in behavior and mood by ADAS-non-Cog (P = 0.008) with 59.2% of the patients being on the mend clinically; and an improvement of 2.4 points by ADL (P = 0.001) with the capacity of ADL improved by at least 10% among 32.75% of the patients. 70% of the patients in huperzine Alpha group scored 1 approximately 3 points, and 27.8% of them scored 1 approximately 2 points by CIBIC-plus. The proportions of patients with an improvement of >/= 4 points by ADAS-Cog were 56.1% and 12.5% in the huperzine Alpha group and placebo group respectively (P = 0.000). The proportions of patients with an improvement of >/= 4 points by MMSE were 37.8% and 10.1% in the huperzine Alpha group and placebo group respectively (P = 0.000). The proportions of patients with an improvement of 1 approximately 3 points in global rating by CIBIC-plus were 59.2% and 40.6% in the huperzine Alpha group and placebo group respectively (P = 0.01). The proportions of patients with an improvement of >/= 10% points by ADL were 32.7% and 17.2% in the huperzine Alpha group and placebo group respectively (P = 0.01). The proportions of patients with an improvement of > 0 points by ADS-non-C0g were 70.0% and 36.3% in the huperzine Alpha group and placebo group respectively (P = 0.000). Mild and transient adverse events (edema of bilateral ankles and insomnia) were observed in 3% of huperzine Alpha treated patients. CONCLUSION: A safe and effective medicine, huperzine Alpha remarkably improves the cognition, behavior, ADL,and mood of AD patients.