Cognition and memory impairment attenuation via reduction of oxidative stress in acute and chronic mice models of epilepsy using antiepileptogenic Nux vomica.

Ethnopharmacological relevance Processed Nux vomica seed extracts and homeopathic medicinal preparations (HMPs) are widely used in traditional Indian and Chinese medicine for respiratory, digestive, neurological and behavioral disorders. Antioxidant property of Nux vomica is well known and recent investigation has highlighted the anticonvulsant potential of its homeopathic formulation. AIM OF THE STUDY: To explore the anticonvulsant and antiepileptogenic potential of Nux vomica HMPs (6CH, 12CH and 30CH potency) in pentylenetetrazole (PTZ) induced acute and chronic experimental seizure models in mice and investigate their effects on cognition, memory, motor activity and oxidative stress markers in kindled animals. MATERIALS AND METHODS: Acute seizures were induced in the animals through 70 mg/kg (i.p.) administration of PTZ followed by the evaluation of latency and duration of Generalized tonic-clonic seizures (GTCS). Subconvulsive PTZ doses (35 mg/kg, i.p.) induced kindling in 29 days, which was followed by assessment of cognition, memory and motor impairment through validated behavioral techniques. The status of oxidative stress was estimated through measurement of MDA, GSH and SOD. RESULTS: HMPs delayed the latency and reduced the duration of GTCS in acute model signifying possible regulation of GABAergic neurotransmission. Kindling was significantly hindered by the HMPs that justified the ameliorated cognition, memory and motor activity impairment. The HMPs attenuated lipid peroxidation by reducing MDA level and strengthened the antioxidant mechanism by enhancing the GSH and SOD levels in the kindled animals. CONCLUSIONS: Nux vomica HMPs showed anticonvulsant and antiepileptogenic potency in acute and chronic models of epilepsy. The test drugs attenuated behavioral impairment and reduced the oxidative stress against PTZ induced kindling owing to which they can be further explored for their cellular and molecular mechanism(s).

Ashwagandha in brain disorders: A review of recent developments.

ETHNOPHARMACOLOGICAL RELEVANCE: Withania somnifera (Family: Solanaceae), commonly known as Ashwagandha or Indian ginseng is distributed widely in India, Nepal, China and Yemen. The roots of plant consist of active phytoconstituents mainly withanolides, alkaloids and sitoindosides and are conventionally used for the treatment of multiple brain disorders. AIM OF THE REVIEW: This review aims to critically assess and summarize the current state and implication of Ashwagandha in brain disorders. We have mainly focussed on the reported neuroactive phytoconstituents, available marketed products, pharmacological studies, mechanism of action and recent patents published related to neuroprotective effects of Ashwagandha in brain disorders. MATERIALS AND METHODS: All the information and data was collected on Ashwagandha using keywords "Ashwagandha" along with "Phytoconstituents", "Ayurvedic, Unani and Homeopathy marketed formulation", "Brain disorders", "Mechanism" and "Patents". Following sources were searched for data collection: electronic scientific databases such as Science Direct, Google Scholar, Elsevier, PubMed, Wiley On-line Library, Taylor and Francis, Springer; books such as AYUSH Pharmacopoeia; authentic textbooks and formularies. RESULTS: Identified neuroprotective phytoconstituents of Ashwagandha are sitoindosides VII-X, withaferin A, withanosides IV, withanols, withanolide A, withanolide B, anaferine, beta-sitosterol, withanolide D with key pharmacological effects in brain disorders mainly anxiety, Alzheimer's, Parkinson's, Schizophrenia, Huntington's disease, dyslexia, depression, autism, addiction, amyotrophic lateral sclerosis, attention deficit hyperactivity disorder and bipolar disorders. The literature survey does not highlight any toxic effects of Ashwagandha. Further, multiple available marketed products and patents recognized its beneficial role in various brain disorders; however, very few data is available on mechanistic pathway and clinical studies of Ashwagandha for various brain disorders is scarce and not promising. CONCLUSION: The review concludes the results of recent studies on Ashwagandha suggesting its extensive potential as neuroprotective in various brain disorders as supported by preclinical studies, clinical trials and published patents. However vague understanding of the mechanistic pathways involved in imparting the neuroprotective effect of Ashwagandha warrants further study to promote it as a promising drug candidate.

The protective effects of in vitro cultivated calculus bovis on the cerebral and myocardial cells in hypoxic mice.

The protective effects of in vitro cultivated calculus bovis (ICCB) on the cerebral and myocardial cells in hypoxic mice and the mechanism were examined. In one group, mice were intragastrically (i.g.) given ICCB for 15 days and then they were subjected to acute cerebral ischemia by decapitation, and then the panting time was recorded. In the other group, 12 min after exposure to hypoxia, mice was administered the ICCB i.g. for 5 days, and then the blood serum and tissues of brain, heart, liver were harvested and examined for SOD, GSH-px and T-AOC activity and content of MDA. The tissues of brain and heart were observed electron-microscopically for ultrastructural changes. The corpus striatum and hippocampus of brain were collected and examined for content of dopamine (DA) and norepinephrine (NE). The ultrastructural examination showed that the pathological change in brain and heart in the ICCB group was very slight, while abnormal changes in the control group were obviously more serious. ICCB significantly prolonged the panting time of the hypoxic mice (P<0.001), increased the activity of SOD, GSH-px, T-AOC in serum and tissues of brain, liver, heart and elevated the content of DA and NE. ICCB also pronouncedly reduced content of MDA in serum and tissues of brain, heart and liver. Significant differences in these parameters were noted between ICCB group and controls. It is concluded that ICCB can exert protective effect on the cells of brain and myocardium by enhancing the tolerance of the tissues to hypoxia and the body's ability to remove free radicals and regulating the neurotransmitters.

Reconstituted mother tinctures of Gelsemium sempervirens L. improve memory and cognitive impairment in mice scopolamine-induced dementia model.

ETHNOPHARMACOLOGICAL RELEVANCE: Gelsemium sempervirens (L.) J.St.-Hil is a herb used for the treatment of various neuroses in both homeopathic and Ayurvedic systems. The present study examines whether Gelsemium reconstituted tincture can protect against scopolamine induced cognitive discrepancies in amnesic mouse model. In order to investigate the protective mechanism of Gelsemium against dementia, in vitro acetyl cholinesterase and ß-secretase enzyme inhibition and estimation of glutathione level in mouse brain were carried out. MATERIALS AND METHODS: The inhibition study on acetyl cholinesterase and ß-secretase enzyme was conducted on brain homogenate supernatant spectrophotometrically using specific substrate. Cognitive enhancement activity was assessed by elevated plus maze and passive avoidance study in scopolamine induced dementia mouse model. Glutathione, an anti-oxidant, was measured spectrophotometrically from scopolamine induced amnesic mice brain supernatant using 5,5'-dithiobis 2-nitrobenzoic acid in the presence and absence of Gelsemium tincture. RESULTS: Significant inhibition was found with Gelsemium on AChE and ß-secretase enzyme with an IC50 of 9.25 and 16.25 µg/ml, respectively, followed by increasing glutathione levels in comparison to the untreated dementia group. The effect of Gelsemium of scopolamine-induced cognitive deficits was determined by measuring the behavioral parameters and the antioxidant status of the brain after scopolamine (1mg/kg i.p.) injected amnesic mice. Gelsemium significantly demonstrated in vivo anti-dementia activity (60% protection) and increased exploratory behavior. CONCLUSION: Our investigations indicated that alkaloid, iridoids and coumarin enriched reconstituted Gelsemium tincture extract displays promising cognitive enhancement in adult mice after short-term oral treatment. Hence, Gelsemium can be a promising anti-dementia agent, mediating the protection against amnesia, attention disorders and learning dysfunctions through dual inhibition of both acetyl cholinesterases (no false positive effect was shown), ß-secretase and antioxidant activity.

Development of thyrotropin-releasing hormone immunoreactivity in the brain of the brown trout Salmo trutta fario.

The development and adult distribution of thyrotropin-releasing hormone-immunoreactive (TRHir) neurons in the brain of the brown trout, Salmo trutta fario, was studied with the streptavidin-biotin immunohistochemical method. Study of embryos, alevin, and juveniles revealed groups of TRHir neurons in the mesencephalon and rhombencephalon that have not been noted previously in adult teleosts. The earliest TRHir cells observed were those of the trigeminal motor nucleus, which expressed this substance only in embryos and alevins. In the forebrain, early-arising TRH populations were observed in the supra- and postcommissural region of the ventral telencephalic area, the anterior parvocellular preoptic nucleus, the organon vasculosum laminae terminalis, the magnocellular preoptic nucleus, the suprachiasmatic nucleus, and the posterior tuberal nucleus. TRHir cells of the olfactory bulb, abundant in the adult, appeared later. A small TRHir neuronal population was transiently observed in the habenula of alevins and juveniles. The laminar nucleus of the mesencephalon contained a small population of TRH cells in alevins and juveniles. In the isthmus, TRH was observed in cells of the interpeduncular nucleus, the nucleus isthmi, the dorsolateral tegmental nucleus, the superior reticular nucleus, and the central gray, although perikarya were TRHir only in alevin and/or juvenile stages. Some vagal motoneurons were TRHir from the late embryo stage onward. TRHir fibers were abundant in several forebrain regions of alevins and juveniles, including the medial region of the dorsal telencephalic area, the ventral telencephalic area and commissural region, the preoptic neuropil, the posterior tubercle, the anterior tuberal nucleus, and the posterior hypothalamic lobe. TRHir fibers invaded the neurohypophysis in early alevins, and their number increased subsequently to adulthood. The parvocellular superficial pretectal nucleus and the optic tectum received a rich TRHir innervation from juvenile stages onward. The interpeduncular nucleus and the secondary gustatory nucleus contained many TRHir fibers. In the rhombencephalon, TRHir fibers were scarce, except in the ventrolateral regions and the inferior olive. The distribution of TRHir fibers suggests that they were mainly related to hypophysiotropic and visceral centers, although the presence of TRH in centers related to the visual system indicates that TRH also plays other roles in the brain. We discuss the possibility that the strong expression of TRH in the embryonic trigeminal motoneurons plays a role in head morphogenesis.

Distribution of calcitonin gene-related peptide-like immunoreactivity in the brain of the small-spotted dogfish, Scyliorhinus canicula L.

The distribution of neuropeptides has been useful in comparing neuronal aggregates of elasmobranchs with those in other vertebrates. The distribution of calcitonin gene-related peptide (CGRP)-like immunoreactivity in the brain of the dogfish was examined with an antiserum to rat alpha-CGRP. Western blot analysis confirms that our antiserum recognizes a single peptide in the dogfish brain very similar to mammalian CGRP. CGRP-like immunoreactivity was located in discrete neuronal groups. CGRP-like-immunoreactive (CGRP-ir) neurons were found in the motor nuclei III, IV, V, VI, VII, IX, and X of the brainstem motor column and in the octavolateral efferent neurons. In the isthmal region, two groups of CGRP-ir neurons appeared in the parabrachial region and reticular substance. Three other CGRP-ir cell groups were observed in the mesencephalon: in the ventral tegmental area, in the substantia nigra, and one widely scattered but numerous population in superficial layers of the optic tectum. In the diencephalon, CGRP-ir cells were observed in the magnocellular preoptic nucleus and the organon vasculosum hypothalami. A population of CGRP-ir cells was also observed in the entopeduncular nucleus in the impar telencephalon. CGRP-ir fibers of central origin were widely distributed in the brain, but the most conspicuous areas were found in the ventral telencephalon, the hypothalamus, the mesencephalic lateral reticular area, and the dorsolateral isthmal region. The neurointermediate lobe of the hypophysis was also richly innervated by CGRP-ir fibers. CGRP-ir sensory fibers of cranial nerves IX and X and of dorsal spinal roots formed very conspicuous terminal fields in the lobus vagi and Cajal's nucleus commissuralis and in the dorsal region of the substantia gelatinosa, respectively. Comparison of the distribution of fibers and perikarya in dogfish and other vertebrates suggests that this CGRP-ir system has been well conserved during evolution.

Development of vimentin and glial fibrillary acidic protein immunoreactivities in the brain of gray mullet (Chelon labrosus), an advanced teleost.

Previous studies in teleosts have revealed the presence of the intermediate filaments vimentin (Vim) and glial fibrillary acidic protein (GFAP) in glial cells of the spinal cord and/or some brain regions, but there is no comprehensive study of their distribution and developmental changes in fishes. Here, the distribution of Vim and GFAP immunoreactivities was studied in the brain of larvae, juveniles, and adults of an advanced teleost, the gray mullet (Chelon labrosus). A different sequence of appearance was observed for expression of these proteins: Vim levels decreased with age, whereas GFAP increased. In general, both immunoreactivities were expressed early in perikarya and endfeet of ependymocytes (tanycytes), whereas expression in radial processes appeared later. In large larvae, the similar expression patterns of Vim and GFAP suggest that some of these glial cells contain both proteins. Subependymal radial glia cells were observed mainly in the optic tectum, exhibiting Vim and GFAP immunoreactivity. The only immunoreactive cells with astrocyte-like morphology were observed in the optic chiasm of the adult, and they were positive for both GFAP and Vim. The perivascular processes of glial cells showed a different distribution of Vim and GFAP during development and had a caudorostral sequence of appearance of immunoreactivities similar to that observed for ependymal and radial glia cells. Several circumventricular organs (the organon vasculosum hypothalami, saccus vasculosus, and area postrema) exhibited highly specialized Vim- and/or GFAP-expressing glial cells. The glial cells of the midline septa of several brain regions were also Vim and/or GFAP immunoreactive. In the adult brain, tanycytes retain Vim expression in several brain regions. As in other vertebrates, the regions with Vim-immunoreactive ventricular and midline glia may represent areas with the capability of plasticity and regeneration in adult brain.

Quantitative distribution of angiotensin II binding sites in rat brain by autoradiography.

Angiotensin II binding sites were localized and quantified in individual brain nuclei from single rats by incubation of tissue sections with 1 nM 125I-[Sar1]-angiotensin II, [3H]-Ultrofilm autoradiography, computerized microdensitometry and comparison with 125I-standards. High angiotensin II binding was present in the circumventricular organs (organon vasculosum laminae terminalis, organon subfornicalis and area postrema), in selected hypothalamic nuclei (nuclei suprachiasmatis, periventricularis and paraventricularis) and in the nucleus tractus olfactorii lateralis, the nucleus preopticus medianus, the dorsal motor nucleus of the vagus and the nucleus tractus solitarii. High affinity (KA from 0.3 to 1.5 X 10(9) M-1) angiotensin II binding sites were demonstrated in the organon subfornicalis, the nucleus tractus solitarii and the area postrema after incubation of consecutive sections from single rat brains with 125I-[Sar1]-angiotensin II in concentrations from 100 pM to 5 nM. These results demonstrate and characterize brain binding sites for angiotensin II of variable high affinity binding both inside and outside the blood-brain barrier.

Localization of angiotensin II receptors along the anteroventral third ventricle area of the rat brain.

Autoradiographic techniques were utilized to localize and to quantify angiotensin II (ANG) binding sites in rat forebrain. Specific, localized ANG binding sites were demonstrated in midline sagittal sections, corresponding to the entire anteroventral third ventricle (AV3V) area, including the nucleus preopticus medianus and the subependymal area of the anterior third ventricle from the nucleus preopticus medianus to the organon vasculosum laminae terminalis. A continuous band of ANG receptors extended dorsally from the nucleus preopticus medianus along the subependymal area of the third ventricle to the organon subfornicalis. Scatchard analysis performed with consecutive sections from single animals revealed a single class of high-affinity ANG receptors in both the organon subfornicalis and the organon vasculosum laminae terminalis. In addition, ANG receptors were localized in areas anatomically and physiologically related to the AV3V area, including the nuclei paraventricularis and periventricularis and the eminentia mediana. These results support the idea that ANG may act as both a hormone and a neurotransmitter in the central regulation of fluid balance and cardiovascular function, and suggest that the circumventricular organs are the most likely sites for an interaction between the peripheral and central ANG systems.

Localization and characterization of atrial natriuretic peptide binding sites in discrete areas of rat brain and pituitary gland by quantitative autoradiography.

Atrial natriuretic peptide [rat (r) ANP6-33 or ANP99-126] binding sites were localized in discrete areas of rat brain and pituitary gland using quantitative autoradiographic techniques. High numbers of rANP6-33 binding sites were concentrated in the circumventricular organs (the organon vasculosum laminae terminalis, organon subfornicalis, and area postrema) and selected hypothalamic nuclei (the nucleus supraopticus, nucleus preopticus medianus and nucleus paraventricularis). High binding was also present in the choroid plexus and the bulbi olfactorii (laminae medullaris interna). A relatively low number of rANP6-33 binding sites was observed in other olfactory, limbic and brainstem areas (the nucleus tractus solitarii, nucleus motoris dorsalis vagii and nucleus hypoglossi), the eminentia mediana and the pituitary gland (anterior and posterior lobes). High-affinity rANP6-33 binding sites were demonstrated in the organon subfornicalis and the area postrema after incubation of consecutive sections from individual rat brains with 125I-rANP6-33 in concentrations from 20 to 400 pM. rANP6-33 binding sites were concentrated in areas associated with angiotensin II and/or vasopressin, suggesting an interaction among these peptides in the central nervous system.

Quantitative determination of angiotensin II binding sites in rat brain and pituitary gland by autoradiography.

Rat brain and pituitary angiotensin II (AII) binding sites were quantitated by incubation of tissue sections with 125I-[Sar1] AII, Ultrofilm radioautography, computerized densitometry, and comparison with 125I-standards at appropriate film exposure times. The highest number of AII binding sites was found in anterior pituitary and the circumventricular organs, organon subfornicalis and organon vasculosum laminae terminalis.

Potentiated antibodies to mu-opiate receptors: effect on integrative activity of the brain.

The effect of homeopathically potentiated antibodies to mu-receptors (10(-100) wt %) on integrative activity of rat brain was studied using the models of self-stimulation of the lateral hypothalamus and convulsions produced by electric current. Electric current was delivered through electrodes implanted into the ventromedial hypothalamus. Single treatment with potentiated antibodies to mu-receptors increased the rate of self-stimulation and decreased the threshold of convulsive seizures. Administration of these antibodies for 7 days led to further activation of the positive reinforcement system and decrease in seizure thresholds. Distilled water did not change the rate of self-stimulation and seizure threshold.

Effect of preparations from potentiated ethanol on the content of biogenic monoamines and metabolism of ethanol in tissues of rats during alcoholization.

The effect of homeopathically potentiated ethanol (C30 and C200) on ethanol metabolism was studied in alcoholized rats. We measured ethanol concentration in the blood, alcohol dehydrogenase activity in the liver, and contents of biogenic monoamines in the hypothalamus, septum, and whole blood. Potentiated preparations of ethanol were efficient after long-term treatment and delayed ethanol elimination from the blood. Preparation C200 increased alcohol dehydrogenase activity. Potentiated preparations of ethanol (particularly C200) produced a positive effect on catecholaminergic and serotoninergic systems of the brain, i.e. they enhanced protective and adaptive reactions.

[Assay of aluminum content in alum and alum-processed medicinal herbs and its distribution in blood and brain in mice].

OBJECTIVE: To assay the aluminum content in alum and some Chinese medicinal herbs processed with alum, and compare the aluminum distribution of these herbs in blood and brain in mice. METHOD: Aluminum content of the herbs was assayed with spectrophotometry, snd aluminum distribution in blood and brain was assayed with atomic absorption spectrophotometry. RESULTS: Aluminum content in alum was found higher than that in the alum-processed herbs. Aluminum distribution in blood and brain after oral administration at equimolecular doses, equivalent to 5.7 mg/kg of metal Al, was studied in mice, and no significant differences were observed. CONCLUSION: Aluminum distribution in blood and brain in mice is related to the dosages of aluminum administered and has little to do with the kind of herbs used. All these medicinal herbs do not improve the absorption of aluminum in mice.

[Experimental study on anti-senility effects of centipede].

Our experiment has shown that the aqueous extract of Centipede can markedly decrease the contents of lipid peroxide in the serum as well as lipofuscin in the liver tissues and brain tissues of rats, enhances the activity of superoxide dismutase in erythrocyte and glutathione in blood, but has no action on the lipofuscin in the heart. In addition, the extract is able to increase the weight of spleen and thymus.

[Biochemical pharmacological effects of lurongjing on young or old mice].

OBJECTIVE: To investigate the effect of Lurongjing (LRJ) on young or old mice. METHODS: In vivo monoamine oxidase (MAO) and superoxide dismutase(SOD) activities, the contents of monoamines(5-HT, NE, DA), malondialdehyde (MDA), RNA and protein, and the incorporations of [3H] Uridine into RNA and [3H] Leucine into protein in liver tissue were measured. RESULTS: The inhibition of MAO-B activity and the increase of monoamines(5-HT, DA) level, the increase of RNA and protein level, and the incorporations of [3H] Uridine into RNA and [3H] Leucine into protein in liver tissue, the enhancement of SOD activity and the decrease of MDA content in brain and liver tissues were selectively observed in old mice(24 months) than in young mice(1 month) by LRJ (i.g., 4.0 g/kg) administrated orally for 10 successive days. CONCLUSION: LRJ exhibits some anti-aging effect in old mice.

[Inhibitory effects of the extract of pilose antler on monoamine oxidase in aged mice].

It was demonstrated that the water extract of Pilose Antler (WEPA) showed a higher inhibitory effect on MAO-B activities in the liver and brain tissues of aged mice, but nearly no effect on NAO-A. WEPA could significantly increase the contents of 5-HT, NE and DA in the brain tissues of aged mice. In vitro experiments revealed that the inhibition of WEPA on MAO-B was competitive, but on MAO-A was of mixed-type.

[Senility-preventing effect of erlingcankang decoction].

Experiments with Erlingcankang Decoction showed that when given to silk worms it could noticeably prolong the growth period of larvae and raise an average of 10-day survival of the male moths; when given to mice it could prolong their life; and when given to on old rats it could markedly raise the contents of SOD in the liver and red cells, lower the content of MAO-B in the brain, LF in the brain and adrenal gland and also LPO in the liver.

[Anti-lipid peroxidaton of the water extract from cornu cervi pantotrichum].

OBJECTIVE: To investigate the anti-lipid peroxidation of the water extract from Cornu Cervi Pantotrichum (WECCP) in vitro. METHODS: The lipid peroxidation (malondialdehyde formation) in microsomes of rat brain, liver and kidney induced by NADPH-Vit C and cyctein-Fe2+, and the superoxide anion radical production (reduced cytochrome c formation) in xanthine-xanthine oxidase system in vitro were evaluated. RESULTS: The lipid peroxidation (MDA formation) in microsomes of rat brain, liver and kidney induced by NADPH-Vit C and cyctein-Fe2+, O2 production(reduced cytochrome c formation) in xanthine-xanthine oxidase system were significantly inhibited by WECCP. CONCLUSION: WECCP exhibits an antioxidant activity.