Effects of autonomic neuropathy on coronary blood flow in patients with diabetes mellitus.

BACKGROUND: C ardiac sympathetic signals play an important role in the regulation of myocardial perfusion. We hypothesized that sympathetically mediated myocardial blood flow would be impaired in diabetics with autonomic neuropathy. METHODS AND RESULTS: We studied 28 diabetics (43+/-7 years old) and 11 age-matched healthy volunteers. PET was used to delineate cardiac sympathetic innervation with [(11)C]hydroxyephedrine ([(11)C]HED) and to measure myocardial blood flow at rest, during hyperemia, and in response to sympathetic stimulation by cold pressor testing. The response to cardiac autonomic reflex tests was also evaluated. Using ultrasonography, we also measured brachial artery reactivity during reactive hyperemia (endothelium-dependent dilation) and after sublingual nitroglycerin (endothelium-independent dilation). Based on [(11)C]HED PET, 13 of 28 diabetics had sympathetic-nerve dysfunction (SND). Basal flow was regionally homogeneous and similar in the diabetic and normal subjects. During hyperemia, the increase in flow was greater in the normal subjects (284+/-88%) than in the diabetics with SND (187+/-80%, P=0.084) and without SND (177+/-72%, P=0.028). However, the increase in flow in response to cold was lower in the diabetics with SND (14+/-10%) than in those without SND (31+/-12%) (P=0.015) and the normal subjects (48+/-24%) (P<0.001). The flow response to cold was related to the myocardial uptake of [(11)C]HED (P<0.001). Flow-mediated brachial artery dilation was impaired in the diabetics compared with the normal subjects, but it was similar in the diabetics with and without SND. CONCLUSIONS: Diabetic autonomic neuropathy is associated with an impaired vasodilator response of coronary resistance vessels to increased sympathetic stimulation, which is related to the degree of SND.

N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP4) impairs neurotransmission in the vas deferens of the rat.

1. The in vitro effects of N-(2-chloroethyl)-N-ethyl-bromobenzylamine (DSP4) were studied in the rat vas deferens. 2. DSP4 inhibited the biphasic motor response induced by field stimulation in a concentration-dependent manner. The concentration of DSP4 that elicited 50% of the maximal inhibition of the twitch response induced by 3 Hz was 10 microM. 3. DSP4 10 microM abolished the motor response induced by exogenously applied noradrenaline and 0.1 mM ATP. Phentolamine (an alpha-adrenoceptor blocker) prevented DSP4 inhibitory effect. 4. DSP4 inhibitory effect was no due to the activation of alpha 2-presynaptic adrenoceptor mechanisms. 5. DSP4 impairs neurotransmission in the rat vas deferens by a postsynaptic alpha 1-adrenoceptor blockade and by an inhibition of the purinergic response.

In vitro effect of thyroxine on cholinergic neurotransmission in rat sympathetic superior cervical ganglion.

This study aimed at examining the effect of thyroid hormones on cholinergic transmission in isolated rat superior cervical ganglia (SCG). In SCG explants incubated with 3H-choline, thyroxine (T4) and 3,3',5-triiodothyronine (T3) added to the medium before a second depolarization stimulus of 60 mM K+ resulted in a dose-dependent increase of S2/S1 ratio for 3H release. The concentration of hormone that produced 50% of maximal increase in K(+)-induced radioactivity release was 8 x 10(-9) M for T4 and 1.6 x 10(-8) M for T3 while 3,3',5,5'-tetraiodothyroacetic acid was almost ineffective. Preincubation of SCG with 10(-7) M iopanoic acid for 30 min before S2, although not affecting by itself S2/S1 ratio, effectively prevented the increase given by T4 or T3. 3H-acetylcholine release by SCG was augmented in a high K+, and the effect was amplified by T4 to a similar extent as that for total 3H release. When added to the incubation medium together with 60 mM K+ for 30 min, T4 (10(-7) M) increased significantly the activity of choline acetyltransferase (ChAT). T4 did not affect ChAT activity in SCG exposed to 4.7 mM K+, nor in SCG homogenates. 3H-choline uptake measured immediately after exposure of SCG to 60 mM K+ decreased by 25%, whereas it increased by 71% after a subsequent 30-min incubation with 4.7 mM K+. Addition of 10(-7) M T4 prevented the changes in choline uptake observed in a high K+ medium. These results indicate that T4 increases SCG cholinergic transmission.

[N-2-chloroethyl-N-ethyl-2-bromobenzylamine (DSP4) increases the liberation of 3H-noradrenaline produced by depolarization of the nerve ending]. / La N-2-cloroetil-N-etil-2-bromobencilamina (DSP4) aumenta la liberacion de 3H-noradrenalina producida por despolarizacion del terminal nervioso.

When injected systemically to rodents, DSP4 inhibits the uptake of noradrenaline (NA) and depletes endogenous NA levels in the central nervous system and in the periphery. Pretreatment with the NA uptake blocker desipramine (DMI), with the NA precursor l-dopa or with the MAD inhibitor, pargyline, prevents the toxic effects of the compound. To investigate the mechanism of the NA depleting action of DSP4, the release of the neurotransmitter induced by nerve stimulation was studied "in vitro" in a tissue sensitive to the neurotoxic action of DSP4 such as the rat cerebral cortex previously loaded with tritiated NA. Incubation with 10 mumol/l DSP4 increased the spontaneous release of tritium from the cortex and produced a two-fold enhancement of tritium outflow during the stimulation of the cortical slices by exposure to K+ 20 mmol/l for 1 m. When the experiments were performed in a Ca++ free medium, DSP4 increased the spontaneous tritium outflow, but did not enhance the NA release by depolarization with K+. This latter effect could be due to the interaction with alpha-adrenoceptors since DSP4 did not potentiate the action of the alpha-antagonist yohimbine (1 mumol/l). DSP4, as did other uptake blockers, reduced the inhibitory effect of clonidine (0.1 mumol/l) on the NA release. Clonidine pretreatment "in vivo" (2 mg/kg), did not counteract the effect of DSP4 (25 mg/kg), suggesting that the enhanced release of NA induced by K+ does not play an important role in the depletion caused by DSP4. This is supported by the fact that DSP4 also enhanced the release of NA evoked by field stimulation of the rat vas deferens, a tissue resistant to the NA depleting action of the compound. The results indicate that the enhancement of the stimulation-induced release caused by DSP4, does not seem to play a triggering role in the NA depletion caused by the compound.

La N-2-cloroetil-N-etil-2-bromobencilamina (DSP4) aumenta la liberacion de 3H-noradrenalina producida por despolarizacion del terminal nervioso. / [N-2-chloroethyl-N-ethyl-2-bromobenzylamine (DSP4) increases the liberation of 3H-noradrenaline produced by depolarization of the nerve ending]

When injected systemically to rodents, DSP4 inhibits the uptake of noradrenaline (NA) and depletes endogenous NA levels in the central nervous system and in the periphery. Pretreatment with the NA uptake blocker desipramine (DMI), with the NA precursor l-dopa or with the MAD inhibitor, pargyline, prevents the toxic effects of the compound. To investigate the mechanism of the NA depleting action of DSP4, the release of the neurotransmitter induced by nerve stimulation was studied [quot ]in vitro[quot ] in a tissue sensitive to the neurotoxic action of DSP4 such as the rat cerebral cortex previously loaded with tritiated NA. Incubation with 10 mumol/l DSP4 increased the spontaneous release of tritium from the cortex and produced a two-fold enhancement of tritium outflow during the stimulation of the cortical slices by exposure to K+ 20 mmol/l for 1 m. When the experiments were performed in a Ca++ free medium, DSP4 increased the spontaneous tritium outflow, but did not enhance the NA release by depolarization with K+. This latter effect could be due to the interaction with alpha-adrenoceptors since DSP4 did not potentiate the action of the alpha-antagonist yohimbine (1 mumol/l). DSP4, as did other uptake blockers, reduced the inhibitory effect of clonidine (0.1 mumol/l) on the NA release. Clonidine pretreatment [quot ]in vivo[quot ] (2 mg/kg), did not counteract the effect of DSP4 (25 mg/kg), suggesting that the enhanced release of NA induced by K+ does not play an important role in the depletion caused by DSP4. This is supported by the fact that DSP4 also enhanced the release of NA evoked by field stimulation of the rat vas deferens, a tissue resistant to the NA depleting action of the compound. The results indicate that the enhancement of the stimulation-induced release caused by DSP4, does not seem to play a triggering role in the NA depletion caused by the compound.

The aziridinium derivative of DSP4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine) accelerates the beating rate of isolated rat atria by enhancing the spontaneous release of noradrenaline.

The aziridinium derivative of the compound N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride (az-DSP4) depletes endogenous noradrenaline stores and exerts neurotoxic actions on noradrenergic neurons. These effects are persistent in the central nervous system and transient in the periphery. To determine if transmitter release plays a role in the noradrenaline depletion caused by az-DSP4, the action of the compound was studied in isolated and spontaneously beating rat atria. 1. az-DSP4 enhanced atrial beating rate when present in the incubation medium at concentrations ranging from 10(-7) M to 10(-4) M but at 10(-3) M decreased that rate below basal levels. 2. Preincubation of atria for 30 min with the noradrenaline uptake blocker desimipramine (DMI, 10(-6) M) or with the beta-blocker propranolol (10(-7) M), abolished the positive chronotropic action of az-DSP4. 3. The rate-accelerating effect of az-DSP4 could be prevented by pretreating the rats with reserpine (5 mg/kg i.p. 24 h) or enhanced by pargyline pretreatment (100 mg/kg i.p. 18 h). 4. az-DSP4 stimulated the spontaneous efflux of tritium from the isolated atria previously labeled with 3H-noradrenaline (4 X 10(-7) M), an increase that was mainly accounted for by DOPEG. 5. COMT and MAO activities in atria homogenates were inhibited by az-DSP4 in a concentration-dependent manner. However, MAO inhibition did not result in a change of the metabolic pattern as could be expected. 6. The results obtained indicate that az-DSP4 enhances the rate of spontaneous beating of isolated rat atria.(ABSTRACT TRUNCATED AT 250 WORDS)

The neurotoxic compound N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride (DSP4) depletes endogenous norepinephrine and enhances release of [3H]norepinephrine from rat cortical slices.

The alkylating compound N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride (DSP4) injected to rodents blocks norepinephrine (NE) uptake and reduces endogenous NE levels in the central nervous system and in the periphery. To investigate the processes leading to these alterations, rat cortical slices were incubated in the presence of DSP4. Cortical NE was depleted by 40% after incubation of slices in 10(-5) M DSP4 for 60 min and this was blocked by desipramine. The spontaneous outflow of radioactivity from cortical slices labeled previously with [3H]NE was enhanced markedly both during exposure to DSP4 and during the subsequent washings, suggesting that NE depletion could be due to this stimulation of NE release. The radioactivity released by DSP4 was accounted for mainly by NE and its deaminated metabolite 3,4-dihydroxyphenylglycol. The enhanced release, independent of external Ca++, apparently originated from the vesicular pool as it was absent after reserpine pretreatment. Activities of the enzymes related to NE synthesis were not altered by DSP4 in vitro and only monoamine oxidase activity was inhibited at high concentrations. Thus, the depletion of endogenous NE produced by DSP4 is probably due to a persistent enhancement of its release from the vesicular pool. Fixation of DSP4 to the NE transport system is necessary but not sufficient to produce the acute NE depletion and the characteristic long-term actions of the compound.