Alterations in free radical scavenging mechanisms following blood-brain barrier disruption.

It has been reported earlier that rat microvessels which constitute the blood-brain barrier (BBB) are rich in free radical scavenging enzymes. In the present investigation, BBB of rat was disrupted by intravenous infusion of the hypertonic saline and changes in enzymes--namely, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR)--were evaluated in the brain microvessels at 30 min after the intravenous administration of hypertonic saline, being the time of peak effect. There was a significant increase in the activities of CAT (40%), GPx (26%), and SOD (16%) over the control values. In addition, within 90 min BBB was found to be reestablished and the levels of enzymes reverted to normal. Malondialdehyde (MDA) levels and activity of lactate dehydrogenase (LDH) remained unaltered during and following disruption, suggesting that there was no change in the membrane lipid environment. Similarly, there was no cell lysis. The results suggest that the disruption of BBB following hypertonic saline administration might be due to an increase in the generation of free radicals in the brain microvessels.

Alterations in the bioantioxidants following thrombosis.

The role of free radicals in rat thrombosis has been demonstrated by studying its scavenging enzyme system. Changes in the bio-antioxidants (GSH-redox cycle, total thiol groups, vitamins E and C) that also offer protection against the free radicals, have, however, not been studied so far. This investigation was undertaken to understand the involvement of these antioxidants during thrombosis. The tissues investigated following thrombosis were blood, platelets, polymorphonuclear leukocytes (PMNLs), heart, and lung. Glutathione (GSH) content in the platelets was observed to be depleted. However, oxidized glutathione (GSSG) contents in the platelets, PMNLs, and blood remain unaltered. In addition, in the whole blood GSH levels were increased significantly, whereas there was no change in the GSSG level. Activity of glutathione reductase (GR) was decreased significantly in platelets and lungs with an increase in the total thiol groups in the lung homogenate. Activity of Glutathione peroxidase (Gpx) remained unaltered in all the tissues studied. In addition, 24% and 15% decrease in the alpha-tocopherol concentration was observed in thrombocytes and PMNLs, respectively, with no change in the ascorbate levels in these cells. Results of this investigation suggest alterations in the GSH-redox cycle in blood, platelets, and lung after thrombosis in the rat.

On the mechanism of potentiation of apomorphine-induced stereotypy due to electroconvulsive shock.

Electroconvulsive shock-induced changes in the intensity of stereotype induced by apomorphine, the binding of [3H]spiroperidol in the corpus striatum, the accumulation of [3H]dopamine in brain and the permeability of the blood-brain barrier, were monitored in rats 30 min after single, or 24 hr after chronic (once daily for 7 days) electroconvulsive shock. There was significant potentiation in stereotypy induced by apomorphine after chronic electroconvulsive shock. The binding of [3H]spiroperidol did not show any change in the affinity (Kd) or density (Bmax) of receptors in the striatum after acute or chronic electroconvulsive shock. The accumulation of dopamine increased significantly in the hypothalamus after acute electroconvulsive shock and in the corpus striatum and hypothalamus after chronic electroconvulsive shock. A significant increase in the entry of sodium fluorescein into the hypothalamus occurred after acute electroconvulsive shock; it increased in all the regions of the brain after chronic electroconvulsive shock. Alteration in the blood-brain barrier (BBB) by electroconvulsive shock leading to increased accumulation of dopamine in the corpus striatum may be responsible for the potentiation of stereotypy.

The mechanism of opening of the blood-brain barrier by hypertonic saline.

The permeability of the blood-brain barrier was assessed in the cat, using Evan's blue as circulant. Regional blood flow and vascular resistance in various areas of brain were measured using radioactive microspheres. Cardiac output (CO), blood pressure (BP), heart rate (HR), pH, pO2 and pCO2 were also measured. Hypertonic saline (5 M, 0.5 ml/kg), administered intravenously, increased the staining of the brain substance. It also produced a marked increase in cerebral blood flow after 5 min and a marked decrease in blood flow to all the regions of the brain after 20 min. The flow returned to normal after 40 min. The vascular resistance decreased at 5 min, increased at 20 min and returned to normal at 40 min. Cardiac output increased significantly at 5 min and decreased at 20 min, while at 40 min it returned to normal. Blood pressure decreased at 5 min, increased at 20 min, while heart rate steadily decreased. A complete recovery of cardiac output, blood pressure and heart rate occurred in 1 hr. No change was observed in pH, pCO2 and pO2. It is concluded that intravenous administration of hypertonic saline causes marked haemodynamic changes and increases the permeability of the blood-brain barrier due to transient impairment of autoregulation.

Anti-inflammatory and some other pharmacological effects of 3,4-trans-2,2-dimethyl-3-phenyl-4-(p-(beta-pyrrolidinoethoxy)-phenyl)-7-methoxy-chroman (Centchroman).

1. Anti-inflammatory, analgesic, antipyretic and other pharmacological activities of a new chroman derivative, Centchroman, have been described.2. Centchroman was found to possess significant anti-inflammatory action in the carrageenin-induced oedema test in mice and rats. It inhibited granuloma formation in the cotton pellet test. The anti-arthritic activity was also evident in formaldehyde-induced arthritis and adjuvant-induced arthritis in rats.3. Centchroman had a lower ulcerogenic index than phenylbutazone.4. The mechanism of the anti-inflammatory action of Centchroman seemed to be independent of endogenous adrenocortical hormones or of its weak oestrogenic activity.5. Centchroman antagonized phenylquinone writhing and bradykinin-induced bronchospasm but was devoid of any antipyretic activity.

Analysis of histamine receptors in the central thermoregulatory mechanism of Mastomys natalensis.

1 The effect of intracerebroventricular (i.c.v.) injection of histamine on the rectal temperature of Mastomys natalensis at ambient temperatures of 10, 24 and 33 degrees C has been studied. 2 Low doses (0.1-1.0 microgram) of histamine produced hypothermia while larger doses (5-20 micrograms) produced dose-dependent hyperthermia. The hypothermic effect was significantly antagonized by mepyramine while the hyperthermia was blocked by cimetidine. 3 Histamine H1-receptor agonists, 2-methyl-histamine and 2-pyridyl-ethylamine, also produced hypothermia which could be blocked by mepyramine. 4 Histamine H2-receptor agonists, impromidine and dimaprit, produced hyperthermia which was antagonized by cimetidine. 5 Pretreatment of the animals with a beta-adrenoceptor antagonist, MJ1999, did not affect the response to histamine. 6 The hyperthermic effect of histamine (10 micrograms) was most marked at 10 degrees C and was attenuated at 33 degrees C. 7 It is concluded that both H1 and H2-histamine receptors are present in the brain of Mastomys. The H1-receptors mediate hypothermia and H2-receptors hyperthermia.

Selective inhibition by glycine of some somatic reflexes in the cat.

1. The effects of intracerebroventricular or intrathecal administration of glycine on some somatic reflexes in chloralosed cats were investigated.2. Glycine produced marked inhibition of the flexor reflex (FR) and crossed extensor reflex (CER) at low doses (2.5-5.0 mg) but even large doses (up to 40.0 mg) had no effect on the facilitation or inhibition of the patellar reflex produced by stimulation of either the reticular formation or the sciatic nerve. The linguomandibular reflex (LMR) could be blocked by 10-20 mg glycine.3. Glycine antagonized strychnine-induced facilitation of FR, CER and LMR and was 5-6 times as active as gamma-aminobutyric acid in this respect.4. The possibility of glycine being a transmitter in spinal pathways is discussed.

Effect of clonidine on the excitability of vasomotor loci in the cat.

1. The effect of clonidine on the direct excitability of hypothalamic, medullary and spinal vasomotor loci has been investigated in cats anaesthetized with chloralose. 2. Clonidine inhibited the excitability of these loci when it was localized to the central sites by intracerebroventricular, intravertebral arterial or intrathecal injection in very low doses (1-2 mug). 3. Topical application of clonidine (0.01 percent and 1.0 percent) to the floor of the fourth ventricle inhibited pressor responses evoked either by stimulation of medullary or hypothalamic vasomotor areas. Inhibition of the pressor responses was accompanied by hypotension and bradycardia in many experiments. 4. It appears that effects of clonidine on the vasomotor loci of the medulla oblongata and the spinal cord contribute to its hypotensive action.

Amplification of platelet response during acute inflammation in rats.

Enhanced aggregation of platelets was observed in platelet-rich plasma, but not in washed platelet suspension (WPS), during acute inflammation in rats. Incubation of WPS with inflammatory plasma increased the aggregatory response to ADP, but the plasma itself did not cause aggregation of platelets. It potentiated the aggregatory response of normal platelets, when platelets were stimulated with arachidonic acid, thrombin, calcium ionophore A23187 or phorbol-12-myristate-13-acetate. Pretreatment of rats with indomethacin (10 mg/kg) did not prevent the increased aggregation response of platelets due to inflammation. This response was not due to any of the biogenic amines nor was it due to platelet factor 4. The activity of the inflammatory plasma was reduced when it was incubated with phospholipase A2, indicating the involvement of platelet-activating factor (PAF). The activity was present both in the lipid and the protein fraction of the inflammatory plasma. The results indicate that a substance(s) released in the circulation during inflammation renders the platelets hyperactive. This substance appears to be a protein which is present in the inflammatory plasma and acts together with PAF to cause increased aggregation of platelets.

Nitrite content and antioxidant enzyme levels in the blood of schizophrenia patients.

RATIONALE: Recent studies have suggested augmentation in the inflammatory response as well as involvement of nitric oxide (NO) in mood disorders. Polymorphonuclear leukocytes (PMN), NO and free radicals have been associated with inflammatory response; however, the status of NO in the PMN has not been investigated so far in schizophrenia patients. OBJECTIVES: The present study was undertaken to investigate levels of nitrite (a metabolite of NO), malonaldehyde (MDA, lipid peroxidation product) and antioxidant enzymes such as superoxide dismutase (SOD), catalase and glutathione peroxidase (Gpx) in the PMN of schizophrenia patients. METHODS: Patients with schizophrenia (n=62) were diagnosed according to DSM-IV and were free of anti-psychotic medications/ECT for at least 3 months. Mean age of the patients was 29.06+/-1.17 years, with a male to female ratio of 4:1, and mean duration of illness was 3.7+/-0.6 years. The control group consisted of 82 healthy subjects with a mean age of 37.0+/-1.26 and a male to female ratio of 5:1. PMN were isolated from the blood. Nitrite, MDA and antioxidant enzymes were estimated by standard biochemical techniques in the PMN of normal healthy controls and schizophrenia patients. Platelet and plasma nitrite levels were also estimated in controls and schizophrenia patients. RESULTS: Nitrite content in the PMN was reduced to 68%, while plasma and platelet nitrite content in schizophrenia patients was not significantly changed in comparison to controls. Malonaldehyde (MDA) content in PMN was significantly augmented in schizophrenia patients but activity of SOD, catalase and Gpx remain unaltered. CONCLUSION: Results obtained indicate a significant decrease in NO synthesis and an increase in MDA in the PMN of schizophrenia patients, while antioxidant enzyme activities were not altered in the PMN of schizophrenia patients. This suggests that the decrease in PMN NO synthesis by PMN might lead to oxidative stress in schizophrenia patients.

Nitric oxide modulates blood-brain barrier permeability during infections with an inactivated bacterium.

The objective of the present investigation was to study the involvement of NO in regulating the permeability of the blood-brain barrier (BBB) during infections, since NOS is known to be induced following infections. The administration of inactivated Escherichia coli (a source of lipopolysaccharide) or poly (I:C), an interferon inducer, to rats increased the permeability of BBB significantly. This increase was found to be potentiated in the presence of L-arginine, a substrate for NOS, while D-arginine had no such effect. N-nitro L-arginine methyl ester, an inhibitor of NOS, and dexamethasone, an inhibitor of NOS induction, blocked the E. coli-induced effects. These results suggest that during infections, NOS inductions causes the release of large quantities of NO, resulting in increased BBB permeability.

Evidence for involvement of nitric oxide in pretectal analgesia in rat.

The study was undertaken to evaluate the role of nitric oxide (NO) in pretectal (PTN)-induced analgesia in rats. Microinjection of varying concentrations of L-arginine (1 nM to 1 microM) produced a quick, long-lasting and concentration-dependent analgesic response, whereas similar concentrations of D-arginine failed to produce analgesia. Moreover pretreatment with N-nitro-L-arginine methyl ester (L-NAME, 1 microM) significantly prevented L-arginine induced analgesia. Further, pretreatment of animals with methylene blue, a known guanylate cyclase inhibitor also prevented the development of analgesia. Our study suggests that L-arginine caused production of NO, which in turn activates pretectal analgesic system involving cyclic GMP.

Novel effects of MPTP: MAO-B unrelated opioidergic activity in mice.

The presence of opioidergic activity after i.p. injection of N-methyl-4-phenyl tetrahydropyridine (MPTP) has been investigated in albino mice by studying analgesia and the Straub reaction. MPTP (6.25-25 mg kg-1) produced a dose-related analgesic response and Straub reaction. These effects of MPTP were effectively antagonized by prior naloxone treatment but remained unaffected after the MAO-B inhibitor deprenyl. MPTP thus possesses significant opioidergic activity and this, unlike its neurotoxic actions, does not appear to be dependent on oxidative conversion to MPP+ (1-methyl-4-phenyl pyridinium).

Cyclo (His-Pro) modulation of body temperature at hot ambient temperature in the desert rat (Mastomys natalensis).

Cyclo(His-Pro) (CHP) has been shown to facilitate cold-induced hypothermia in the desert rat Mastomys natalensis. In the present study, we examined the role of endogenous CHP in hyperthermia induced by hot ambient temperature (40 degrees C) in the above rodent species. The results of these studies show that housing rodents at 40 degrees C resulted in a altered distribution of CHP in the brain, with a rise in hypothalamic content accompanied by an increase in rectal temperature. While administration of exogenous CHP decreased hyperthermia, immunoneutralization of endogenous CHP increased hyperthermia. The results of these studies show that changes in endogenous CHP levels may affect body temperature regulation.

Role of endogenous cyclo(His-Pro) in cold-induced hypothermia in the desert rat (Mastomys natalensis).

Central administration of exogenous cyclo(His-Pro) (CHP) is known to produce hypothermia in rodents. In the present study, we examined the role of endogenous CHP in cold-induced hypothermia in the desert rat, Mastomys natalensis. The results of these studies show that a rise in hypothalamic CHP content accompanied a decrease in rectal temperature during cold exposure. Immunoneutralization of endogenous CHP resulted in a significant decline in cold-induced hypothermia. In addition, central administration of cyclo(Ala-Gly), a structural analogue of CHP, also led to a decrease in cold-induced hypothermia. The results of these studies show that changes in endogenous CHP levels may affect body temperature regulation.

Attenuation of alcohol-induced hypothermia by cyclo (His-Pro) and its analogs.

Acute administration of cyclo (His-Pro) to rats cause a dose-dependent decrease in ethanol-induced hypothermia. Bromination of the imidazole moiety of histidine in cyclo (His-Pro) resulted in a significant increase in its potency to attenuate ethanol hypothermia. In contrast, benzylation of the imidazole moiety of histidine or the substitution of one or both of the amino acids in cyclo(His-Pro) led to a total loss of its thermomodulatory activity. In conclusion, it appears from these preliminary data that it may be possible to design analogs of CHP that may be effective antagonists for ethanol hypothermia.

Permeability function related to cerebral microvessel enzymes during ageing in rats.

Cerebral microvessels from rats were prepared and characterized by their enrichment of specific markers, namely alkaline phosphatase (AP) and tau-glutamyl transpeptidase (tau-GT). Further, it was observed that AP and tau-GT registered marked increase in aged rats. On the contrary, lactate dehydrogenase (LDH) activity decreased with the increasing age. Monoamine oxidase A activity in the microvessels decreased with age whereas MAO-B moved in the reverse direction. No noticeable change was seen in acetyl-cholinesterase activity with increasing age of rats.

Endothelin antagonizes the hypotension and potentiates the hypertension induced by clonidine.

Modification of clonidine-induced cardiovascular effects by endothelin-1 (ET-1) was studied in male Sprague-Dawley rats. A dose-dependent decrease in blood pressure and heart rate was produced by clonidine (100, 250 and 500 micrograms/kg i.v.). Lower doses produced only a fall in blood pressure (through central alpha-adrenoceptors) while higher doses of clonidine produced an initial hypertensive response (through peripheral alpha-adrenoceptors) and subsequent longer lasting hypotension and bradycardia. The hypotension and bradycardia induced by 100 and 250 micrograms/kg i.v. dose of clonidine were completely blocked by ET-1 (100 ng/kg i.v.) pretreatment. Conversely, the hypertensive response induced by high dose of clonidine (500 micrograms/kg i.v.) was significantly potentiated by ET-1 pretreatment. In cervical sectioned rats, i.v. administered clonidine failed to produce any hypotensive effect, indicating lack of central effect of clonidine. ET-1 significantly (P < 0.0005) potentiated the hypertensive response of a low dose (50 micrograms/kg i.v.) of clonidine in cervical-sectioned rats. I.c.v. administration of clonidine (1, 2, 4 and 6 micrograms) produced a dose-dependent decrease in blood pressure and heart rate. ET-1 pretreatment (25 ng i.c.v.) transiently blocked the clonidine-induced decrease in blood pressure and heart rate for about 10 min but the hypotension and bradycardia was observed subsequently. Since the major site of action of clonidine is the ventral surface of medulla, clonidine was applied directly to the ventral surface of medulla and produced a decrease in blood pressure and heart rate.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of molsidomine and free radical scavengers on the pulmonary thromboembolism in mice.

Molsidomine is a prodrug of nitric oxide (NO) and is used in the treatment of coronary heart disease. Recent studies with NO have shown that it can exert beneficial as well as toxic effects. It was, therefore, considered desirable to evaluate the effect of this drug in an animal model of thrombosis. It was administered to mice, one hour prior to the thrombotic challenge, in a dose range of 1 to 100 mg/kg ip. In lower doses (1-30 mg/kg), it offered protection by 24-38%, and at 50 and 100 mg/kg it protected 54% and 58% animals respectively. Mice pretreated with molsidomine (3, 10 and 30 mg/kg) and free radical scavengers (superoxide dismutase, catalase, mannitol and deferoxamine, suboptimal doses) together offered significantly more protection against thrombosis than either one of them alone. The results suggest that a combination of molsidomine and free radical scavengers should be more effective than molsidomine alone in the treatment of thrombosis and other ischemic disorders.

Status of antioxidants in brain microvessels of monkey and rat.

Levels of certain antioxidants namely reduced glutathione (GSH), ascorbic acid (Vit C), alpha-tocopherol (Vit E) and antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) were compared in monkey and rat brain microvessels which constitute the blood-brain barrier (BBB). The BBB of both the species contains appreciable amounts of the antioxidants to protect against oxidative damage. The level of protection in rat seems to be more efficient than monkey since rat microvessels contain higher concentrations of some of the bio-antioxidants. The comparative status of enzymatic and non-enzymatic protective system against oxidation in the brain microvessels has been discussed.