RESUMEN
Hyperforin, a lipophilic constituent of medicinal herb St. John's Wort, has neurobiological effects including antidepressant activity, antibiotic potency, anti-inflammatory activity and anti-tumoral properties. Furthermore, hyperforin activates transient receptor potential conical channel-6 (TRPC6), a nonselective cation channel. To elucidate the roles of hyperforin and TRPC6 in neuroinflammation in vivo, we investigated the effect of hyperforin on neuroinflammatory responses and its related events in the rat piriform cortex (PC) following status epilepticus (SE). Hyperforin attenuated microglial activation, p65-serine 276 NFκB phosphorylation, and suppressed TNF-α expression in the PC following SE. Hyperforin also effectively alleviated SE-induced vasogenic edema formation, neuronal damage, microglial TRPC6 induction and blood-derived monocyte infiltration. Our findings suggest that hyperforin may effectively attenuate microglia-mediated neuroinflammation in the TRPC6-independent manner.
Asunto(s)
Microglía/efectos de los fármacos , Floroglucinol/análogos & derivados , Corteza Piriforme/efectos de los fármacos , Estado Epiléptico/patología , Terpenos/farmacología , Factor de Transcripción ReIA/metabolismo , Animales , Electroencefalografía , Técnica del Anticuerpo Fluorescente , Masculino , Floroglucinol/farmacología , Fosforilación , Corteza Piriforme/metabolismo , Ratas , Ratas Sprague-Dawley , Estado Epiléptico/inducido químicamenteRESUMEN
Recently, it has been reported that astroglial loss/dysfunction plays a role in epileptogenesis. In addition, astroglial loss is accompanied by up-regulation of P2X7 receptor expression in microglia. Therefore, we investigated whether P2X7 receptor is involved in astroglial damages induced by status epilepticus (SE). In the present study, astroglial loss showed the regional-specific manner and the differential responses to P2X7 receptor functions. Both OxATP and brilliant blue G (P2X7 receptor antagonists) infusion prevented apoptotic astroglial loss in the molecular layer of the dentate gyrus and the frontoparietal cortex, while it promoted clasmatodendrosis in the CA1 region as compared to saline treatment. In contrast, BzATP (a P2X7 receptor agonist) treatment exacerbated apoptotic astroglial loss in the molecular layer of the dentate gyrus and the frontoparietal cortex, but alleviated SE-induced astroglial swelling in the CA1 region. Astroglial loss in the piriform cortex was not affected by P2X7 receptor agonist- or antagonist-infusion. These findings suggest that P2X7 receptor function differently modulates SE-induced astroglial loss in distinct brain regions.
Asunto(s)
Astrocitos/patología , Región CA1 Hipocampal/patología , Dendritas/efectos de los fármacos , Giro Dentado/patología , Lóbulo Frontal/patología , Lóbulo Parietal/patología , Agonistas del Receptor Purinérgico P2X/farmacología , Antagonistas del Receptor Purinérgico P2X/farmacología , Receptores Purinérgicos P2X7/fisiología , Estado Epiléptico/patología , Adenosina Trifosfato/análogos & derivados , Adenosina Trifosfato/farmacología , Animales , Apoptosis/efectos de los fármacos , Apoptosis/fisiología , Astrocitos/efectos de los fármacos , Región CA1 Hipocampal/efectos de los fármacos , Forma de la Célula/efectos de los fármacos , Dendritas/ultraestructura , Giro Dentado/efectos de los fármacos , Lóbulo Frontal/efectos de los fármacos , Masculino , Microglía/efectos de los fármacos , Microglía/patología , Lóbulo Parietal/efectos de los fármacos , Pilocarpina/toxicidad , Ratas , Ratas Sprague-Dawley , Receptores Purinérgicos P2X7/efectos de los fármacos , Colorantes de Rosanilina/farmacología , Estado Epiléptico/inducido químicamente , Estado Epiléptico/fisiopatologíaRESUMEN
AIM: To observe neuroprotective effects of raw and roasted licorice against hypoxia and ischemic damage. METHODS: When elucidating the protective effects of raw and roasted licorice, we analyzed the lactate dehydrogenase (LDH) release using PC12 cells after hypoxia in an in vitro study and after transient forebrain ischemia in an in vivo study on Mongolian gerbils. RESULTS: Raw and roasted licorice significantly reduced LDH release from PC12 cells exposed to an hypoxic chamber for 1 h. In the roasted licorice-treated group, the decrease of LDH release was more pronounced compared to that of the raw licorice-treated group. In roasted licorice-treated animals, approximately 66%-71% of CA1 pyramidal cells in the ischemic hippocampus were stained with cresyl violet compared to the control group. However, in the raw licorice-treated animals, no significant neuroprotection against ischemic damage was shown. In addition, ischemic animals in roasted licorice-treated group maintained the Cu, Zn-superoxide dismutase (SOD1) activity and protein levels compared to the control group, while in raw licorice-treated group SOD1 activity and protein levels were reduced significantly. High pressure liquid chromatography analysis showed that non-polar compounds containing glycyrrhizin-degraded products, such as glycyrrhetinic acid (GA) and glycyrrhetinic acid monoglucuronide (GM), were increased in roasted licorice. CONCLUSION: Roasted licorice had neuroprotective effects against ischemic damage by maintaining the SOD1 levels. In addition, the difference in protective ability between raw and roasted licorice may be associated with non-polar compounds, such as GA and GM.
Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Glycyrrhiza , Ataque Isquémico Transitorio/metabolismo , Fármacos Neuroprotectores/farmacología , Superóxido Dismutasa/metabolismo , Animales , Hipoxia de la Célula , Medicamentos Herbarios Chinos/aislamiento & purificación , Gerbillinae , Glycyrrhiza/química , Hipocampo/metabolismo , Hipocampo/patología , Ataque Isquémico Transitorio/patología , L-Lactato Deshidrogenasa/metabolismo , Masculino , Fármacos Neuroprotectores/aislamiento & purificación , Células PC12 , Plantas Medicinales/química , Células Piramidales/efectos de los fármacos , Ratas , Superóxido Dismutasa-1RESUMEN
This article investigates the long-term effects of soybean isoflavones (ISO) on the changes of calbindin D-28k (CB) immunoreactivity in the hippocampus in middle-aged ovariectomized female rats as well as middle-aged control female and male rats to identify any correlation between calcium and phytoestrogens. In the CA1 region, CB immunoreactivity in the ovariectomized females was similar to that of the control females, whereas CB immunoreactivity in the males was significantly lower than that of the control females. In the dentate gyrus, CB immunoreactivity in the ovariectomized females and males was significantly lower than that of the control females. CB immunoreactivity in all groups was increased dose-dependently after ISO treatment in the CA1 region and dentate gyrus. This result suggests that ISO treatment enhances the expression of CB immunoreactivity in the hippocampus in the middle-aged rats.
Asunto(s)
Regulación de la Expresión Génica/efectos de los fármacos , Hipocampo/efectos de los fármacos , Isoflavonas/administración & dosificación , Proteína G de Unión al Calcio S100/metabolismo , Proteínas de Soja/administración & dosificación , Factores de Edad , Animales , Calbindinas , Relación Dosis-Respuesta a Droga , Femenino , Inmunohistoquímica/métodos , Masculino , Ovariectomía/métodos , Ratas , Ratas Sprague-DawleyRESUMEN
In the present study, we studied the neuroprotective effects of berberry extract (BE) against ischemic damage and the temporal and spatial alterations of N-methyl-D-aspartate receptor type 1 (NR1) and NR2A/2B immunoreactivities in the gerbil hippocampal CA1 region after transient ischemia to examine anti-ischemic effects and its role in transient forebrain ischemia. In the vehicle-treated group, the percentage of cresyl violet positive pyramidal cells in the CA1 region was about 11.4% compared to the sham-operated group 4 d after ischemic insult. BE showed neuroprotective effects against ischemic damage after ischemia-reperfusion. In the BE-treated groups, about 60-75% of CA1 pyramidal cells were stained with cresyl violet 4 d after ischemic insult. We observed the percentage of berberine (7.45+0.85 mg/g in BE) by HPLC, which is active ingredient of BE. NR1 immunoreactivity in the stratum pyramidale of the CA1 region in the vehicle-treated group was significantly increased at 30 min after transient forebrain ischemia, while at this time the NR1 immunoreactivity in the BE-treated groups was significantly low compared to the vehicle-treated group. The pattern of NR2A/B immunoreactivity in the stratum pyramidale of the BE-treated group and its protein levels were similar to that in the vehicle-treated group after ischemic insult. These results suggest that BE has potent neuroprotective effects against ischemic damage via the reduction of NR1 activity.
Asunto(s)
Berberis/química , Hipocampo/metabolismo , Hipocampo/patología , Ataque Isquémico Transitorio/metabolismo , Ataque Isquémico Transitorio/patología , Neuronas/patología , Fármacos Neuroprotectores/farmacología , Receptores de N-Metil-D-Aspartato/metabolismo , Animales , Benzoxazinas , Berberina/análisis , Berberina/farmacología , Western Blotting , Muerte Celular/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Gerbillinae , Inmunohistoquímica , Indicadores y Reactivos , Masculino , Oxazinas , Extractos Vegetales/farmacología , Estándares de ReferenciaRESUMEN
We cloned and expressed human pyridoxal-5\'-phosphate (PLP) phosphatase, the coenzymatically active form of vitamin B6, in Escherichia coli using pET15b vector. Monoclonal antibodies (mAb) were generated against purified human brain PLP phosphatase in mice, and four antibodies recognizing different epitopes were obtained, one of which inhibited PLP phosphatase. The binding affinities of these four mAbs to PLP phosphatase, as determined using biosensor technology, showed that they had similar binding affinities. Using the anti-PLP phosphatase antibodies as probes, we investigated their cross-reactivities in various mammalian and human tissues and cell lines. The immunoreactive bands obtained on Western blots had molecular masses of ca. 33 kDa. Similarly fractionated extracts of several mammalian cell lines all produced a single band of molecular mass 33 kDa. We believe that these PLP phosphatase mAbs could be used as valuable immunodiagnostic reagents for the detection, identification, and characterization of various neurological diseases related to vitamin B6 abnormalities.
Asunto(s)
Anticuerpos Monoclonales/química , Encéfalo/enzimología , Monoéster Fosfórico Hidrolasas/química , Monoéster Fosfórico Hidrolasas/metabolismo , Animales , Bioquímica/métodos , Técnicas Biosensibles , ADN Complementario/metabolismo , Mapeo Epitopo , Epítopos/química , Escherichia coli/metabolismo , Biblioteca de Genes , Vectores Genéticos , Humanos , Inmunohistoquímica/métodos , RatasRESUMEN
The 80% aqueous MeOH extracts from the root of Angelica dahurica, found to inhibit the activities of GABA degradative enzymes GABA transaminase (GABA-T) and succinic semialdehyde dehydrogenase (SSADH), were fractionated using EtOAc, n-BuOH and H2O. Repeated column chromatography for the EtOAc and n-BuOH fractions led to the isolation of two new coumarins, oxypeucedanin hydrate-3''-butyl ether and isopraeroside IV along with six known coumarins, isoimperatorin, imperatorin, phellopterin, oxypeucedanin hydrate, nodakenin and 3'-hydroxymarmesinin, and two polyacetylenes, falcarindiol and octadeca-1,9-dien-4,6-diyn-3,8,18-triol. Of the isolated pure compounds, imperatorin and falcarindiol inactivated the GABA-T activities in both time- and concentration-dependent manners. The kinetic studies showed that imperatorin and falcarindiol reacted with the GABA-T with a second-order rate constant of 2.3 +/- 0.2 mm(-1) min(-1) and 1.5 +/- 0.1 mm(-1) min(-1), respectively. It is postulated that imperatorin and falcarindiol are able to elevate the neurotransmitter GABA levels in the central nervous system by an inhibitory action on the GABA degradative enzyme GABA-T.
Asunto(s)
4-Aminobutirato Transaminasa/antagonistas & inhibidores , Angelica , Cumarinas/farmacología , Glucósidos/farmacología , Fitoterapia , Extractos Vegetales/farmacología , Animales , Encéfalo/enzimología , Bovinos , Cumarinas/administración & dosificación , Cumarinas/química , Cumarinas/uso terapéutico , Glucósidos/administración & dosificación , Glucósidos/química , Glucósidos/uso terapéutico , Infusiones Intravenosas , Extractos Vegetales/administración & dosificación , Extractos Vegetales/química , Extractos Vegetales/uso terapéutico , Raíces de Plantas , Succionato-Semialdehído Deshidrogenasa/antagonistas & inhibidoresRESUMEN
In the present study, we investigated the chronological alterations in SOD1 and its copper chaperone (chaperone for superoxide dismutase, CCS) immunoreactivities and their neuroprotective effects against neuronal damage in the gerbil hippocampus after 5 min of transient forebrain ischemia. SOD1 and CCS immunoreactivities were significantly increased in the stratum pyramidale of the CA1 region at 24 and 12 h after ischemic insult, respectively. At 24 h after ischemic insult, the SOD1 and CCS immunoreactivities were colocalized in the CA1 pyramidal cells of the stratum pyramidale. Thereafter, their immunoreactivities were significantly decreased in the CA1 region. To elucidate the effects of CCS or CCS/SOD1, we constructed the expression vectors PEP-1-SOD and PEP-1-CCS. In the CCS-treated group and the CCS/SOD1-treated group, 43.9 and 78.9% pyramidal cells, respectively, compared to the sham-operated group, were stained with cresyl violet 5 or 7 days after ischemic insult. The distribution pattern of active astrocytes and microglia in the PEP-CCS/SOD1-treated group 5 days after ischemic insult was similar to that of the sham-operated group. In addition, the SOD activity in the PEP-CCS- or PEP-CCS/SOD1-treated group was maintained by 10 days after ischemic insult. The SOD activity was higher in the PEP-CCS/SOD1-treated group vs the CCS-treated group. These results suggest that the enhanced expression of SOD1 and CCS may be related to compensatory mechanisms against ischemic damage and that cotreatment with CCS and SOD1 has a greater neuroprotective effect than treatment with CCS or SOD1 in isolation.
Asunto(s)
Isquemia Encefálica/patología , Hipocampo/patología , Chaperonas Moleculares/metabolismo , Neuronas/patología , Superóxido Dismutasa/metabolismo , Animales , Astrocitos/metabolismo , Astrocitos/patología , Western Blotting , Isquemia Encefálica/metabolismo , Isquemia Encefálica/prevención & control , Gerbillinae , Hipocampo/metabolismo , Inmunohistoquímica , Masculino , Microglía/metabolismo , Microglía/patología , Fármacos Neuroprotectores/metabolismo , Proteínas Recombinantes de FusiónRESUMEN
Grape seed extract (GSE) possess cardioprotective abilities by functioning as in vivo antioxidants and by virtue of their ability to directly scavenge ROS including hydroxyl and peroxyl radicals. In the present study, we investigated the neuroprotective effects of grape seed extract (GSE) in the gerbil hippocampus after 5 min transient forebrain ischemia. Neuronal cell density in GSE-treated ischemic animals was significantly increased as compared with vehicle-treated ischemic animals 4 days after ischemic insult. In the GSE-treated groups, about 60% of pyramidal cells of the sham-operated group were stained with cresyl violet 4 days after ischemic insult. In this study, we found that GSE had neuroprotective effects on neuronal injury by inhibiting DNA damage in the CA1 region after ischemia. In vehicle-treated groups, 8-hydroxy-2'-deoxyguanosine (8-OHdG) immunoreactivity was significantly changed time-dependently, whereas the immunoreactivity in the GSE-treated group was similar to the sham-operated group. In addition, we confirmed that astrocytes and microglia did not show significant activation in the CA1 region 4 days after ischemia-reperfusion, because many CA1 pyramidal cells were not damaged. Therefore, these results suggest that GSE can protect ischemic neuronal damage by inhibiting DNA damage after transient forebrain ischemia.
Asunto(s)
Daño del ADN/efectos de los fármacos , Desoxiguanosina/análogos & derivados , Hipocampo/fisiología , Neuronas/metabolismo , Semillas/química , Vitis/química , 8-Hidroxi-2'-Desoxicoguanosina , Animales , Antioxidantes/farmacología , Astrocitos/metabolismo , Benzoxazinas , Western Blotting , Isquemia Encefálica , Recuento de Células , Gerbillinae , Inmunohistoquímica , Masculino , Microglía/metabolismo , Neuronas/efectos de los fármacos , Fármacos Neuroprotectores/farmacología , Oxazinas , Extractos Vegetales/farmacología , Células Piramidales/metabolismoRESUMEN
To investigate the protective activity of soy isoflavones on neurons, the effects of isoflavones on cholinergic enzyme activity, immunoreactivities of cholinergic enzyme, and delayed matching-to-place (DMP) performance were measured in normal elderly rats. Male Sprague-Dawley rats (n = 48; 10 mo old) were assigned to 3 groups: CD (control diet), ISO 0.3 (0.3 g/kg soy isoflavones diet), and ISO 1.2 (1.2 g/kg soy isoflavones diet). After 16 wk of consuming these diets, choline acetyltransferase (ChAT) activity in the ISO 0.3 group was greater in cortex and basal forebrain (BF; P < 0.05) than in controls. In BF, ChAT activity was also significantly greater in the ISO 1.2 group than in control rats. Acetylcholine esterase (AChE) activity in the ISO 0.3 group was significantly inhibited in cortex, BF, and hippocampus and in the ISO 1.2 group in cortex and hippocampus. Choline acetyltransferase immunoreactivity (ChAT-IR) in the ISO 1.2 group was significantly greater than in controls in the medial septum area. ChAT-IR in the ISO 0.3 and ISO 1.2 groups was significantly higher than in the CD group in the hippocampus CA1 area. Spatial DMP performance by the ISO 0.3 group showed significantly shorter swimming time than by the CD group. These findings show that soy isoflavones can influence the brain cholinergic system and reduce age-related neuron loss and cognition decline in male rats.
Asunto(s)
Dieta , Isoflavonas/uso terapéutico , Memoria/efectos de los fármacos , Actividad Motora/efectos de los fármacos , Proteínas de Soja/uso terapéutico , Envejecimiento , Animales , Encéfalo/efectos de los fármacos , Encéfalo/enzimología , Encéfalo/patología , Colina O-Acetiltransferasa/metabolismo , Isoflavonas/administración & dosificación , Masculino , Ratas , Ratas Sprague-Dawley , Proteínas de Soja/administración & dosificaciónRESUMEN
Pyridoxal phosphatase catalyzes the dephosphorylation of pyridoxal 5'-phosphate (PLP) and pyridoxine 5'-phosphate. A human brain cDNA clone was identified to the PLP phosphatase on the basis of peptide sequences obtained previously. The cDNA predicts a 296-amino acid protein with a calculated Mr of 31698. The open reading frame is encoded by two exons located on human chromosome 22q12.3, and the exon-intron junction contains the GT/AG consensus splice site. In addition, a full-length mouse PLP phosphatase cDNA of 1978 bp was also isolated. Mouse enzyme encodes a protein of 292 amino acids with Mr of 31512, and it is localized on chromosome 15.E1. Human and mouse PLP phosphatase share 93% identity in protein sequence. A BLAST search revealed the existence of putative proteins in organism ranging from bacteria to mammals. Catalytically active human PLP phosphatase was expressed in Escherichia coli, and characteristics of the recombinant enzyme were similar to those of erythrocyte enzyme. The recombinant enzyme displayed Km and kcat values for pyridoxal of 2.5 microM and 1.52 s(-1), respectively. Human PLP phosphatase mRNA is differentially expressed in a tissue-specific manner. A single mRNA transcript of 2.1 kb was detected in all human tissues examined and was highly abundant in the brain. Obtaining the molecular properties for the human PLP phosphatase may provide new direction for investigating metabolic pathway involving vitamin B6.
Asunto(s)
Monoéster Fosfórico Hidrolasas/biosíntesis , Monoéster Fosfórico Hidrolasas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Encéfalo/metabolismo , Cromosomas Humanos Par 15 , Clonación Molecular , ADN Complementario/metabolismo , Bases de Datos como Asunto , Electroforesis en Gel de Poliacrilamida , Escherichia coli/metabolismo , Exones , Humanos , Intrones , Cinética , Ratones , Modelos Genéticos , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Péptidos/química , Monoéster Fosfórico Hidrolasas/química , ARN Mensajero/metabolismo , Proteínas Recombinantes/química , Homología de Secuencia de Aminoácido , Distribución Tisular , Vitamina B 6/metabolismoRESUMEN
The succinic semialdehyde dehydrogenase (SSADH) inhibitory component was isolated from the EtOAc fraction of Lactuca sativa through repeated column chromatography; then, it was identified as phytol, a diterpenoid, based on the interpretation of several spectral data. Incubation of SSADH with the phytol results in a time-dependent loss of enzymatic activity, suggesting that enzyme modification is irreversible. The inactivation followed pseudo-first-order kinetics with the second-rate order constant of 6.15 x 10(-2) mM(-1) min(-1). Complete protection from inactivation was afforded by the coenzyme NAD+, whereas substrate succinic semialdehyde failed to prevent the inactivation of the enzyme; therefore, it seems likely that phytol covalently binds at or near the active site of the enzyme. It is postulated that the phytol is able to elevate the neurotransmitter GABA levels in central nervous system through its inhibitory action on one of the GABA degradative enzymes, SSADH.