RESUMO
Alzheimer's disease (AD) is associated with damage to hippocampal neurons and declines in cognitive functions. The accumulation of amyloid peptides is regarded as a crucial event in the initiation of AD. The neurotoxicity induced by Aß25-35 peptides was used to screen for cytoprotective factors in vitro, and the cognitive deficits induced by the injection of Aß25-35 into the hippocampus were used to evaluate effect on learning and memory. Our previous study revealed that hydrolysate of polygalasaponins (HPS) clearly improve the cognitive deficits induced by the injection of Aß25-35 in mice, but the potential active constituent of HPS remains unclear. The purposes of this study were to separate and purify the secondary saponins of HPS, screen for neuroprotective effects of the constituents in vitro, and to evaluate the effect of cognition in vivo. Various chromatographic methods were used to separate and purify the HPS. The neuroprotective effects were examined in Aß25-35-damage-induced PC12 cells. The protective effect of tenuifolin on the cognitive impairments induced by Aß25-35 injection was assessed using the Morris water maze and step-through passive avoidance tests. Tenuifolin and fallaxsaponin A were isolated from the HPS. Tenuifolin possessed neuroprotective effects against Aß25-35-induced apoptosis in PC12 cells and significantly improved the cognitive deficits induced by the intrahippocampal injection of Aß25-35 in mice. Thus, tenuifolin is one of the active constituents of HPS against the neurotoxicity induced by Aß25-35 peptides in vitro and in vivo.
Assuntos
Peptídeos beta-Amiloides , Diterpenos do Tipo Caurano , Fármacos Neuroprotetores , Neurotoxinas , Fragmentos de Peptídeos , Animais , Humanos , Camundongos , Ratos , Doença de Alzheimer/tratamento farmacológico , Doença de Alzheimer/etiologia , Doença de Alzheimer/psicologia , Peptídeos beta-Amiloides/fisiologia , Diterpenos do Tipo Caurano/isolamento & purificação , Diterpenos do Tipo Caurano/farmacologia , Hipocampo/efeitos dos fármacos , Hipocampo/fisiopatologia , Hidrólise , Técnicas In Vitro , Aprendizagem/efeitos dos fármacos , Memória/efeitos dos fármacos , Modelos Animais , Fármacos Neuroprotetores/isolamento & purificação , Fármacos Neuroprotetores/farmacologia , Neurotoxinas/toxicidade , Células PC12 , Fragmentos de Peptídeos/fisiologia , Fitoterapia , Saponinas/química , Saponinas/farmacologiaRESUMO
Polygalasaponins are the major active constituents of Polygala tenuifolia exhibiting antiamnesic activity, but their applications are limited due to their toxicities. Evidence showed that the toxicities can be attenuated by hydrolysis. Herein, effects of a hydrolysate of polygalasaponins (HPS) on cognitive impairment induced by Aß(25-35) were assessed by Morris water maze and step-through passive avoidance tests. The impaired spatial reference memory was improved by HPS (50 and 100 mg/kg). In the acquisition trial of step-through test, HPS (50 and 100 mg/kg) increased the latency into the dark chamber and decreased the error frequency significantly (P < .05). However, no significant change was observed during the retention trial. Additionally, HPS increased the corresponding SOD activities (62.34%, 22.09%) and decreased MDA levels (28.21%, 32.35%) in both cortex and hippocampus as compared to model animals. These results show that HPS may be a useful treatment against amnesia probably via its antioxidant properties.
RESUMO
OBJECTIVE: To study the inhibitive effects of an effective section of a prescription of traditional Chinese medicine (TCM-ES) on influenza virus A FM1 strain in vitro. METHODS: The experiments were performed by microcytopathic-inhibiting-assay, Neutral Red stain and inhibiting plaque-forming units (PFU) test on MDCK cell strain. By means of observing the cytopathic effects (CPE), measuring the absorbance [D(lambda)] and counting the PFU, according to Reed-Muench assay, the TCM-ES's effective dosage of 50 percentage (EC50) and treatment index (TI) to FM1 were calculated. The inhibiting dose of 50 percentage of PFU (IC50) was also figured up. RESULTS: By CPE assay, TCM-ES'S EC50, MTC and TI to 100TCID50 FM1 strain infection were (300 +/- 18.3) mg/L, (75 +/- 6.8) mg/L and (7.1 +/- 0.7), respectively; Whereas, ribavirin's EC50, MTC and TI was (52.3 +/- 10.1) mg/L, (25 +/- 4.1) mg/L and (20.8 +/- 5.1), respectively. By Neutral Red stain assay,TCM-ES's IC50 and TI was (285.0 +/- 19.2) mg/L and (7.2 +/- 0.6), respectively; whereas ribavirin's IC50 and TI was (45.3 +/- 4. 9) mg/L and (21.2 +/- 3.1), respectively. By reducing PFU assay, the IC50 of TCM-ES and ribavirin was 300 mg/L and 50 mg/L, respectively. All the results above were almost consistent with each other (P > 0.05). CONCLUSIONS: TCM-ES assumes antiviral action on IFV-FM1 strain in a certain degree in vitro and can rebel intracellular virus. But it is worse than the positive control medicine of ribavirin and is worthy of further study.
Assuntos
Antivirais/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Vírus da Influenza A/efeitos dos fármacos , Plantas Medicinais/química , Animais , Antivirais/isolamento & purificação , Células Cultivadas , Cães , Combinação de Medicamentos , Medicamentos de Ervas Chinesas/isolamento & purificação , Vírus da Influenza A/fisiologia , Concentração Inibidora 50 , RNA Viral/efeitos dos fármacos , Ribavirina/farmacologia , Replicação Viral/efeitos dos fármacosRESUMO
Paeonol, a phenolic component from the root bark of Paeonia moutan, is traditionally used as a Chinese herbal medicine to activate the blood flow and remove blood stasis. Evidence shows that paeonol have anti-tumor, anti-inflammatory, and analgesic effects; however, the underlying mechanisms remain unknown. In this study, we investigated the molecular mechanisms by which paeonol exerts the anti-tumor effects by using a murine model of hepatoma established by in vivo injection of mouse HepA-hepatoma cells. Treatment of mice with 100, 200, or 400 mg/kg/day of paeonol significantly inhibited the growth of the HepA tumor in mice, induced HepA cell apoptosis as demonstrated by light microscopy and electron microscopy analyses, decreased the expression of Bcl-2 and increased the expression of Bax in HepA tumor tissues in a dose-related manner. Administration of paeonol in vivo also elevated serum levels of IL-2 and TNF-alpha in tumor-bearing mice. Moreover, splenocytes and macrophages isolated from paeonol-treated HepA tumor-bearing mice produced higher levels of IL-2 and TNF-alpha in response to concanavalin A and lipopolysaccharide stimulation, respectively, compared to these isolated from non-treated HepA tumor-bearing mice. In vitro treatment with paeonol was able to directly stimulate IL-2 and TNF-alpha production in splenocytes and macrophages from tumor-bearing mice, respectively. In conclusion, paeonol has the anti-tumor effect against hepatoma cells, which are likely mediated via induction of tumor cell apoptosis and stimulation of IL-2 and TNF-alpha production. Paeonol could be a promising drug to treat hepatocellular carcinoma.
Assuntos
Acetofenonas/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Interleucina-2/metabolismo , Neoplasias Hepáticas Experimentais/tratamento farmacológico , Fator de Necrose Tumoral alfa/metabolismo , Acetofenonas/uso terapêutico , Animais , Antineoplásicos Fitogênicos/uso terapêutico , Linhagem Celular Tumoral , Células Cultivadas , Relação Dose-Resposta a Droga , Interleucina-2/sangue , Neoplasias Hepáticas Experimentais/imunologia , Neoplasias Hepáticas Experimentais/patologia , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/imunologia , Camundongos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Baço/efeitos dos fármacos , Baço/imunologia , Fator de Necrose Tumoral alfa/sangue , Regulação para Cima , Proteína X Associada a bcl-2/metabolismoRESUMO
A rice gene, OsPIPK 1, encoding a 792-aa putative phosphatidylinositol 4-phosphate 5-kinase (PIPK), was identified and characterized. Comparison between the cDNA and genomic sequences revealed the presence of 10 exons (39-1050 bp) and 9 introns (88-745 bp) in OsPIPK 1 gene. The deduced amino acid sequence of OsPIPK1 contains a lipid kinase domain that is highly homologous to those of previously isolated PIPKs, and structural analysis revealed the intriguing presence of multiple MORN motifs at the N-terminus. The MORN motifs have also been detected in PIPKs from Arabidopsis thaliana and Oryza sativa, but not in the well-characterized PIPKs from animal and yeast cells. RT-PCR analysis indicated that OsPIPK 1 was expressed almost constitutively in roots, shoots, stems, leaves and flowers, and up-regulated following treatment with plant hormones or application of various stresses. An antisense transgenic strategy was used to suppress the expression of OsPIPK 1, and homozygous transgenic plants showed earlier heading (7-14 days earlier) than control plants, suggesting that OsPIPK 1 negatively regulates floral initiation. This was further confirmed by morphologic observation showing earlier floral development in antisense plants, as well as leaf emergence measurement indicating delayed leaf development under OsPIPK 1 deficiency, a common phenotype observed with earlier flowering. RT-PCR analysis and cDNA chip technology were used to examine transcripts of various genes in the transgenic plants and the results showed altered transcriptions of several flowering-time or -identity related genes, suggesting that OsPIPK 1 is involved in rice heading through regulation of floral induction genes, signaling and metabolic pathways.