Acute administration of nicotine impairs the hypotensive responses to bradykinin in rats.

Nicotine may contribute to smoking-induced endothelial dysfunction because of its ability to impair endothelium-dependent vasodilatation. We investigated whether the acute administration of nicotine changes the hypotensive responses to bradykinin in rats. The effects of pre-treatment with losartan or enalapril on the nicotine-induced changes in the responses to bradykinin were also evaluated. In study 1, anesthetized rats were cannulated via carotid artery for the measurement of mean arterial pressure. Dose-response curves to bradykinin (0.1, 0.4, 1.6, 6.4, 25 and 100 microg/kg) were generated before and 10 min after the injection of nicotine (200 microg/kg, i.v.) or saline. The individual dose-response curves were fitted to a four-parameter logistic equation using the ALLFIT program, which provided an estimate of the maximal response (E(max)) and of the dose of bradykinin producing the half-maximal response (ED(50)). In study 2, rats were pre-treated orally with losartan (10 mg/kg/day) or enalapril maleate (25 mg/kg/day) for 2 weeks. Control rats received tap water alone. After pre-treatment, the rats were anesthetized and used as described in study 1. Nicotine decreased the E(max) (from 73.0+/-7.5 to 65.7+/-3.3 mm Hg; P<0.05) but did not affect the ED(50). In study 2, losartan or enalapril did not affect nicotine-induced decrease in responses to bradykinin; E(max) decreased in both groups (from 68.7+/-6.3 to 62.8+/-4.2 mm Hg, and from 53.8+/-13.0 to 43.1+/-7.1 mm Hg, respectively; P<0.05) without significantly changing the ED(50). These results suggest that nicotine impairs the endothelium-dependent hypotensive responses to bradykinin. This effect is not influenced by inhibition of the angiotensin-converting enzyme or by blockade of the angiotensin AT(1) receptors.

Intracerebroventricular administration of canatoxin to rats increases the circulating levels of luteinizing hormone.

Recent evidence has implicated the central nervous system as a target organ for canatoxin, a toxic protein present in Canavalia ensiformis seeds. This toxin activates the lipoxygenase pathway of arachidonic acid metabolism and can thus induce the release of substances mediated by lipoxygenase products. In the present study, the circulating levels of luteinizing hormone (LH) were measured by RIA in male Wistar rats (200-240 g) after the administration of canatoxin into the lateral cerebral ventricle. Canatoxin (0.5-2 micrograms in 2 microliters daily for 3 days) caused a dose- and time-dependent increase in the plasma levels of LH. The total dose of canatoxin used is subconvulsive. At 2, 4 and 24 h after 2 micrograms of canatoxin LH levels were increased by 10%, 43% and 61%, respectively, compared to vehicle-injected animals (0.18 +/- 0.03 ng/ml). This response to 2 micrograms of canatoxin was not attenuated by pretreatment with two different lipoxygenase inhibitors, nordihydroguaiaretic acid (NDGA, 125 mg/kg) or esculetin (ECLT, 125 mg/kg), ip, 1 h before each canatoxin (CNTX) injection; % increase in LH with CNTX alone: 61%; CNTX+NDGA: 54%; CNTX+ECLT:76%; N = 5/group. These data show that intracerebral injection of CNTX in rats increases circulating levels of LH via a mechanism that is independent of the lipoxygenase pathway.

Intracerebroventricular administration of canatoxin to rats increases the circulating levels of luteinizing hormone

Recent evidence has implicated the central nervous system as a target organ for canatoxin, a toxic protein present in Canavalia ensiformis seeds. This toxin activates the lipoxygenase pathway of arachidonic acid metabolism and can thus induce the release of substances mediated by lipoxygenase products. In the present study, the circulating levels of luteinizing hormone (LH) were measured by RIA in male Wistar rats (200-240 g) after the administration of canatoxin into the lateral cerebral ventricle. Canatoxin (0.5-2 micrograms in 2 microliters daily for 3 days) caused a dose- and time-dependent increase in the plasma levels of LH. The total dose of canatoxin used is subconvulsive. At 2, 4 and 24 h after 2 micrograms of canatoxin LH levels were increased by 10 per cent , 43 per cent and 61 per cent , respectively, compared to vehicle-injected animals (0.18 +/- 0.03 ng/ml). This response to 2 micrograms of canatoxin was not attenuated by pretreatment with two different lipoxygenase inhibitors, nordihydroguaiaretic acid (NDGA, 125 mg/kg) or esculetin (ECLT, 125 mg/kg), ip, 1 h before each canatoxin (CNTX) injection; per cent increase in LH with CNTX alone: 61 per cent ; CNTX+NDGA: 54 per cent ; CNTX+ECLT:76 per cent ; N = 5/group. These data show that intracerebral injection of CNTX in rats increases circulating levels of LH via a mechanism that is independent of the lipoxygenase pathway

Increased insulin circulating levels induced by canatoxin in rats.

Canatoxin, a proteic neurotoxin from Canavalia ensiformis seeds, raises circulating insulin levels and induces hypoglycemia in rats. The hyperinsulinemia produced by canatoxin (6 micrograms kg-1, 20 min; 108% of increase in female rats) was both time and dose dependent and lasted only about 30 min, while the fall in blood glucose levels (around 30%) was long lasting. The hyperinsulinemic response to canatoxin was greater in females (+108 +/- 18%) than males (+43 +/- 8%), but no difference was noted in the hypoglycemia. Pretreatment of rats with either naloxone, naltrexone, atropine or hexamethonium abolished both the hyperinsulinemia and the hypoglycemia. These data suggest that these phenomena are influenced by opioids and depend on parasympathetic stimulation.

Further studies on the hypoxia produced by canatoxin in rats.

Canatoxin, a convulsant neurotoxin from the seeds of Canavalia ensiformis, induces lipoxygenase-dependent hypoxia in rats which is blocked by hexamethonium. The purpose of the present study was to examine the relationship between canatoxin-induced hypoxia and bronchoconstriction. Since several effects of the toxin are very similar to those described for morphine and opioid-like peptides, the effects of opioid antagonists were also investigated. Pretreatment of male, adult Wistar rats (200-250 g) with cyproheptadine (80 micrograms/kg, ip, N = 6) and isoproterenol (100 micrograms/kg, ip, N = 6) partially blocked (% variation of pO2: CNTX alone: -26.67 +/- 2.56, N = 6; with cyproheptadine: -16.15 +/- 2.97*, N = 6; with isoproterenol: -17.73 +/- 1.93*, N = 6; *P < 0.05 as compared to CNTX alone) the hypoxia but no effect was observed with diphenhydramine (2 mg/kg, ip, N = 6) or atropine (2 mg/kg, ip, N = 6). The hypoxemic effect of canatoxin (100 micrograms/kg, i.v., 20 min, N = 6) was also almost completely blocked with either naloxone (1 mg/kg, sc, N = 6) or naltrexone (5 mg/kg, sc, N = 6). The results presented here provide evidence suggesting that both opioid peptides and bronchoconstriction seem to play a role in the hypoxia caused by canatoxin.

Further studies on the hypoxia produced by canatoxin in rats

Canatoxin, a convulsant neurotoxin from the seeds of Canavalia ensiformis, induces lipoxygenase-dependent hypoxia in rats which is blocked by hexamethonium. The purpose of the present study was to examine the relationship between canatoxin-induced hypoxia and bronchoconstriction. Since several effects of the toxin are very similar to those described for morphine and opioid-like peptides, the effects of opioid antagonists were also investigated. Pretreatment of male, adult Wistar rats (200-250 g) with cyproheptadine (80 ug/kg, ip, N=6) and isoproterenol (100 ug/kg, ip, N+6) partially blocked (% variation of pO2:CNTX alone: -26.67 ñ 2.56, N=6; with cyproheptadine: 16.15 ñ 2.97*, N=6; with isoproterenol: 17.73 ñ 1.93*, N=6; *P<0.05 as compared to CNTX alone) the hypoxia but no effect was observed with diphenhydramine (2 mg/kg, ip, N=6) or atrophine (2 mg/kg, ip, N=6). The hypoxemic effect of canatoxin (100 ug/kg iv 20 min, N=6) was also almost completely blocked with either naloxone (1 mg/kg, sc, N=6) or naltrexone (5 mg/kg, sc, N=6). The results presented here provide evidence suggesting that both opioid peptides and bronchoconstriction seem to play a role in the hypoxia caused by canatoxin

Convulsions induced by canatoxin in rats are probably a consequence of hypoxia.

Canatoxin, a convulsant neurotoxin from the seeds of Canavalia ensiformis, induces lipoxygenase-dependent hypoxia and sex-related alterations of carbohydrate metabolism in rats which are blocked by glucose, diazepam and hexamethonium. The present study analyzes the possible casual relationship between the convulsant action of canatoxin and its effects on carbohydrate metabolism. The incidence of canatoxin-induced convulsions was greater in male than in female rats. Pretreatment of male rats with drugs that block hypoxia, such as glucose (2.5 g/kg, iv, 15 min), diazepam (5 mg/kg, ip, at 48 h, 24 h and 15 min), hexamethonium (4 mg/kg, ip, 15 min) and NDGA (125 mg/kg, ip, 1 h), also protected the animals against convulsions, respiratory distress and death. These results suggest that canatoxin-induced convulsions are probably the consequence of hypoxia and both effects are mediated by lipoxygenase activation.

Alterations in rat carbohydrate metabolism induced by canatoxin as a probable consequence of primary hypoxia.

1. Canatoxin, a protein displaying lipoxygenase-activating properties isolated from Canavalia ensiformis seeds, induces hypoxia and hyperglycemia in male rats. 2. Liver glycogen, blood glucose and lactate levels were measured in male and female rats after canatoxin (50 mU, iv) injection. Increased levels of serum glutamic oxaloacetic transaminase activity were used as an indicator of hepatic injury. 3. There was no sex-related difference observable during canatoxin-induced hypoxia but male and female rats did show different patterns of metabolic change and hepatic injury after toxin administration. Increased blood glucose and lactate levels, liver glycogenolysis and hepatic injury were observed in male rats while female rats showed only hypoglycemia and glycogenolysis. 4. Pretreatment of male rats with either glucose, diazepam or hexamethonium abolished both the hypoxia and hepatic injury and the metabolic alterations produced by toxin injection. 5. The results suggest that the metabolic alterations and hepatic injury detected after canatoxin injection may be a consequence of primary hypoxia.

Alterations in rat carbohydrate metabolism induced by canatoxin as a probable consequence of primary hypoxia

Canatoxin, a protein displaying lipoxygenase-activating properties isolated from Canavalia ensiformis seeds, induces hypoxia and hyperglycemia in male rats. Liver glycogen, blood glucose and lactate levesls were measured in male and female rats after canatoxin (50 mU, iv) injection. Increased levels of serum glutamic oxaloacetic transaminase activity were used as an indicator of hepatic injury. There was no sex-related difference observable during canatoxin-induced hypoxia but male and female rats did whow different patterns of metabolic change and hepatic injury after toxin observed in male rats while female rats showed only hypoglycemia and glycogenolysis. Pretreatment of male rats with either glucose, diazepam or hexamethonium abolished both the hypoxia and hepatic injury and the metabolic alterations produced by toxin injection. The results suggest that the metabolic alterations and hepatic injury detected after canatoxin injection may be a consequence of primary hypoxia

Convulsions induced by canatoxin in rats are probably a consequence of hypoxia

Canatoxin, a convulsant neurotoxin from the seeds of Canavalia ensiformis, induces lipoxygenase-dependent hypoxia and sex-related alterations of carbohydrate metabolism in rats which are blocked by glucose, diazepam and hexamethonium. The present study analyzes the possible causal relationship between the convulsant action of canatoxin and its effects on carbohydrate metabolism. The incidence of canatoxin-induced convulsions was greater in male than in female rats. Pretreatment of male rats with drugs that block hypoxia, such as glucose (2.5 g/kg,iv,15 min), diazepam (5 mg/kg,ip, at 48 h, 24 animals against convulsions, respiratory distress and death. These results suggest that canatoxin/induced convulsions are probably the consequence of hypoxia and both effects are mediated by lipoxygenase activation