Alterations in the nitric oxide synthase binding sites and non-adrenergic, non-cholinergic mediated smooth muscle relaxation in the diabetic rabbit bladder outlet: possible relevance to the pathogenesis of diabetic cystopathy.
OBJECTIVE
To investigate the effect of diabetes mellitus (DM) on the density and distribution of nitric oxide synthase (NOS) and the smooth muscle responses to non-adrenergic, non-cholinergic (NANC) nerve stimulation and exogenous nitric oxide (NO) in the rabbit lower urinary tract.


METHODS
Transverse sections of detrusor, bladder neck and urethra, from control and six months alloxan-induced DM New Zealand White rabbits were incubated with a radioligand for NOS ([3H]-L-N(G)-nitroarginine). Densitometric analysis was performed on the autoradiographs. NADPH diaphorase histochemistry was also used as a marker for NOS activity. Responses to NANC nerve stimulation (5 to 20 Hz) and to NO (10(-6) to 3x10(-4) M.) on smooth muscle strips from detrusor, bladder neck and urethra were measured in organ baths.


RESULTS
NOS binding sites were significantly (p<0.03) more dense in the bladder neck than in the detrusor in both DM and control groups. In DM bladder neck, NOS binding sites were significantly (p<0.04) increased compared with the controls. NADPH diaphorase activity appeared markedly increased in the detrusor, bladder neck and urethra of DM animals compared with controls. The mean IC50 for exogenous NO in control versus DM were not statistically different in the bladder neck (1.03x10(-4) M versus 9.8x10(-5) M) and urethra (8.1x10(-5) M versus 8.8x10(-5) M), but the relaxations to 5x10(-6) M of NO were significantly impaired (p<0.04) in the DM urethral smooth muscle. NANC nerve-mediated relaxations were significantly impaired (p<0.001) in the DM urethral smooth muscle.


CONCLUSIONS
Alterations of both the NOS binding sites and functional responses to NANC nerve stimulation suggest that NO may have a pathophysiological role in the urinary bladder dysfunction associated with DM.
Department of Urology, Royal Free Hospital, London, United Kingdom.
F H
FH
Mumtaz
M E
ME
Sullivan
C S
CS
Thompson
M R
MR
Dashwood
K M
KM
Naseem
K R
KR
Bruckdorfer
D P
DP
Mikhailidis
R J
RJ
Morgan
D000818:Animals; D001345:Autoradiography; D001665:Binding Sites; D003921:Diabetes Mellitus, Experimental / Q000150:complications / Q000201:enzymology* / Q000503:physiopathology*; D008297:Male; D009126:Muscle Relaxation / Q000502:physiology*; D009130:Muscle, Smooth / Q000201:enzymology* / Q000503:physiopathology*; D009252:NADPH Dehydrogenase / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D019335:Nitroarginine; D011817:Rabbits; D014521:Urethra / Q000201:enzymology / Q000503:physiopathology; D001743:Urinary Bladder / Q000201:enzymology* / Q000503:physiopathology*; D001745:Urinary Bladder Diseases / Q000201:enzymology* / Q000209:etiology / Q000503:physiopathology*
D019335:Nitroarginine; D019001:Nitric Oxide Synthase; D009252:NADPH Dehydrogenase