Recent evidence implicates NO (Nitric oxide) as the principal mediator in an erection. To investigate the role of NO in the human erectile function, we studied the distribution pattern of nitroxergic fibers in the corpus cavernosum specimens obtained from 38 men using nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry. Diffusely scattered delicate nerve fibers showing blue color reaction after NADPH-d histochemical staining were observed in normal control specimens from potent men. The neurogenic impotence group showed a statistically-significant decrease in the number of positive fibers compared to the normal control group. The number of positive fibers in the non-neurogenic impotence group was decreased compared to the normal control group but was statistically insignificant. With nitric oxide synthase (NOS) immunohistochemical stain, immunoreactive nerve bundles were easily seen in normal control specimens from potent men. NOS immunoreactive nerve bundles were contained within the corpus cavernosa which stained with NADPH-d reaction. Our results suggest that nitric oxide, a potent smooth muscle relaxing neurotransmitter in the autonomic nervous system, plays a physiologic role in erectile function and NADPH-d enzyme histochemical staining on the biopsied corpus cavernosum may be used as an important diagnostic method in the evaluation of neurogenic impotence.
Histocytochemistry ; Human ; Impotence/enzymology* ; Male ; NADPH Dehydrogenase/metabolism ; Nitric-Oxide Synthase/metabolism* ; Penis/enzymology* ; Tissue Distribution

Reactive oxygen species (ROS) are thought to contribute to the progression of cardiovascular disease (CVD). At present, it has been known that the NADH/NADPH oxidase system is the major source of superoxide in the vascular system. Cytochrome b-245 (P22phox), which is a critical component of NADH/NADPH oxidase, plays an important role in electron transport and producing the superoxide anion. It is considerable to take attention to whether the polymorphism of P22phox gene is associated with a risk of coronary heart disease (CHD). To distinguish the relationship between them will be beneficial to elucidate the genetic mechanisms of CHD, and develop a new genetic marker to provide theoretical base for the prevention and cure of CHD.
Coronary Disease ; genetics ; Genetic Markers ; Humans ; Membrane Transport Proteins ; genetics ; NADPH Dehydrogenase ; genetics ; NADPH Oxidases ; Phosphoproteins ; genetics ; Polymorphism, Genetic ; Risk Factors

Nitric oxide(NO) is a free radical which serves a wide variety of functions on vascular tone, neurotransmission, immune cytotoxicity, and many others. Nitric oxide synthase(NOS) is the biosynthetic enzyme of NO and colocalized with NADPH diaphorase(NADPH-d) activity in many tissues. The author aimed to assess the changes that occur in this populations of neurons in the streptozotocin-induced diabetic rat where the retinal vasculature is known to be dysfunctional. The 8 rats was a diabetic group and the other 8 was a control group. Diabetes was induced with a single intraperitoneal injection of streptozotocin(65mg/kg). Four weeks later, the retina was flat mounted and stained with NADPH-d. Counting of the stained cells was made. There was a 20.6% decrease in the total number of positively staining cells in the retinas of the diabetic group(2532+/-192) compared with those of the control group(3188+/-176)(p<0.001). It is worth to suggest the close correlation between NO released from retinal neurons and the microcirculatory dysfunction in diabetic retinopathy.
Animals ; Diabetic Retinopathy ; Injections, Intraperitoneal ; NADP* ; NADPH Dehydrogenase* ; Neurons ; Nitric Oxide ; Rats* ; Retina* ; Retinal Neurons ; Retinaldehyde* ; Synaptic Transmission

PURPOSE: The purpose of this study was to investigate the changes of serum testosterone, intracavernous pressure, expression of nitric-oxide synthase (NOS) and content of penile smooth muscle in aged rat. MATERIALS AND METHODS: Sprague Dawley rats were used for this study and divided into control and aging groups. Intracavernous pressure was measured by stimulating the cavernous nerve with 10volt, 2.4mA. Expression of NOS was measured by immunohistochemical staining for NADPH (nicotinamide adenine dinucleotide phosphate) diaphorase and the content of penile smooth muscle was measured by Masson's trichrome staining for corpus cavernosum. In each case the stained area-tissue ratio was calculated by computer scanning. RESULTS: Compared with the control group (3.34-0.25ng/ml), the serum testosterone level of the aged group (1.41-0.37ng/ml) was decreased significantly. Compared with the control group (106.7-13.2cmH2O), the intracavernosal pressure of the aged group (71.2-12.3cmH2O) was decreased significantly. Immunohistochemical staining for NOS showed that NADPH diaphorase was stained as brown nerve fiber. Compared with the control group (40.5-3.1%), NOS activity of the aged group (9.5-2.5%) was decreased significantly. On Masson's trichrome staining, the content of penile smooth muscle was 62.8 3.9% in the control group. Compared with the control group, the smooth muscle content of the aged group (35.2-2.4%) was decreased significantly. CONCLUSIONS: In conclusion, the factors involved in erectile function were decreased in aged rats.
Adenine ; Aging* ; Animals ; Muscle, Smooth ; NADP ; NADPH Dehydrogenase ; Nerve Fibers ; Nitric Oxide Synthase* ; Rats* ; Rats, Sprague-Dawley ; Testosterone

PURPOSE: We performed immunohistochemical analysis of vasoactive intestinal polypeptide (VIP) and nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase in the anterior vaginal wall, and we investigated their relations to the females sexual life and their stress urinary incontinence. MATERIALS AND METHODS: From December 2002 to April 2003, 55 urinary incontinent women, who were treated with tension-free vaginal tape (TVT), participated in this study. Their average age was 52.3 years old. We evaluated their sexual function with the Korean version of female sexual function index (FSFI). Anterior vaginal wall tissues 1x1cm in size were obtained during the TVT operation, and they were analyzed by immunohistochemical technique for VIP and NADPH diaphorase. We counted the number of nerve fibers containing VIP or NADPH diaphorase in the microscopic field of view. We verified the results with a Student's t-test and spearman test to identify the relations immunohistochemical results to the females sexual function and urinary incontinence. RESULTS: Expression of VIP was significantly low in grade III incontinence, but there was not a significant difference for the other parameters of incontinence. Expression of NADPH diaphorase had no significant relation with any factor of incontinence. For the relation between expression of VIP and NADPH diaphorase and the FSFI score, the domain of arousal shows a significant difference with the expression of VIP and NADPH diaphorase, according to FSFI score. CONCLUSIONS: From the above results, we suggest that VIP and NADPH diaphorase may affect the structure and functions of the female pelvic floor and these neurotransmitters act on the arousal phase of female sexual function.
Arousal ; Female* ; Humans ; NAD* ; NADP* ; NADPH Dehydrogenase ; Nerve Fibers ; Neurotransmitter Agents ; Niacinamide* ; Pelvic Floor ; Sexuality ; Suburethral Slings ; Urinary Incontinence* ; Vasoactive Intestinal Peptide*

PURPOSE: We performed immunohistochemical analysis of vasoactive intestinal polypeptide (VIP) and nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase in the anterior vaginal wall, and we investigated their relations to the females sexual life and their stress urinary incontinence. MATERIALS AND METHODS: From December 2002 to April 2003, 55 urinary incontinent women, who were treated with tension-free vaginal tape (TVT), participated in this study. Their average age was 52.3 years old. We evaluated their sexual function with the Korean version of female sexual function index (FSFI). Anterior vaginal wall tissues 1x1cm in size were obtained during the TVT operation, and they were analyzed by immunohistochemical technique for VIP and NADPH diaphorase. We counted the number of nerve fibers containing VIP or NADPH diaphorase in the microscopic field of view. We verified the results with a Student's t-test and spearman test to identify the relations immunohistochemical results to the females sexual function and urinary incontinence. RESULTS: Expression of VIP was significantly low in grade III incontinence, but there was not a significant difference for the other parameters of incontinence. Expression of NADPH diaphorase had no significant relation with any factor of incontinence. For the relation between expression of VIP and NADPH diaphorase and the FSFI score, the domain of arousal shows a significant difference with the expression of VIP and NADPH diaphorase, according to FSFI score. CONCLUSIONS: From the above results, we suggest that VIP and NADPH diaphorase may affect the structure and functions of the female pelvic floor and these neurotransmitters act on the arousal phase of female sexual function.
Arousal ; Female* ; Humans ; NAD* ; NADP* ; NADPH Dehydrogenase ; Nerve Fibers ; Neurotransmitter Agents ; Niacinamide* ; Pelvic Floor ; Sexuality ; Suburethral Slings ; Urinary Incontinence* ; Vasoactive Intestinal Peptide*

PURPOSE: This study examined the changes in intracavernous pressure, expression of nitric oxide synthase(NOS), and content of penile smooth muscle in castrated rats and testosterone-supplied castrated rats. MATERIALS AND METHODS: Sprague Dawley rats were used for this study and divided into control, castrated, and testosterone-supplied castrated groups. Castration was performed by bilateral orchietomy under general anesthesia, and testosterone propionate 3 mg/kg was injected subcutaneously daily for a week beginning 4 weeks after orchiectomy. Intracavernous pressure was measured by stimulating the cavernous nerve at 10 volts, 2.4 mA. Expression of NOS was measured by immunohistochemical staining for NADPH diaphorase, and content of penile smooth muscle was measured by H&E staining of the corpus cavernosum. The stained area-to-tissue ratio was calculated by computer scanning for each case. RESULTS: Compared with the control group(3.45+/-0.25 ng/ml), the serum testosterone level of the castrated group (0.78+/-0.34 ng/ml) was lower. Serum testosterone level was restored in the testosterone-supplied castrated group. Compared with the(67.2+/-14.3 cmH2O) was decreased (p <0.05). There was no significant difference between the testosterone-supplied group(94.7+/-11.4 cmH2O) and control group, so intracavernosal pressure was restored by testosterone treatment. Immunohistochemical staining for NOS showed that NADPH diaphorase was stained as brown nerve fiber. Compared with the control group(37.5+/-2.8%), the NOS activity of the castrated group(7.5+/-2.1%) was significantly decreased(p <0.05). NOS activity was slightly increased in the testosterone-supplied group(47.5+/-2.4%) compared with the control group, but the difference was not statistically significant. Thus, testosterone treatment restored NOS activity after castration. By H&E staining, the content of penile smooth muscle was 76.5+/-2.8% in the control group, but significantly lower in the castrated group(46.2+/-3.4% p <0.05). Smooth muscle content was slightly decreased in the testosterone-supplied group(63.8+/-4.7%) compared with control group, but the difference was not statistically significant. Thus, smooth muscle content was restored by testosterone treatment after castration. CONCLUSIONS: Decline of factors involved in erectile function can be restored by testosterone replacement after castration.
Anesthesia, General ; Animals ; Castration ; Male ; Muscle, Smooth ; NADPH Dehydrogenase ; Nerve Fibers ; Nitric Oxide ; Orchiectomy ; Penile Erection* ; Rats* ; Rats, Sprague-Dawley ; Testosterone Propionate ; Testosterone*

Recently, it is well established that nitric oxide synthase (NOS) produces nitric oxide (NO), which is known to act as an important neural mediator of smooth muscle relaxation in various organs. The present study was undertaken to investigate the role played by NO in relaxing bladder outlet by correlating its action with the existence, distribution and activity of NOS. The experiments consisted of nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase staining for the identification of NOS containing nerve fibers, NADPH diaphorase activity with spectrophotometric assay for NOS catalytic activity, Western blot analysis with polyclonal antibrain NOS antibody for the expression of neuronal NOS, and in vivo isovolumetric bladder contraction (IBC) and reflex urethral .relaxation (RUR) for the physiologic role of NO. On NADPH diaphorase histochemistry in the rat, NADPH positive staining was localized in neurons innervating the lower genitourinary tract including the urinary bladder and the proximal urethra. NADPH positive nerve fibers were mainly found in urethral area, whereas they were less common in detrusor. In assay of NADPH diaphorase activity on various organs of the rat, the NOS activity regionally predominated in the cerebellum, proximal urethra, and urinary bladder in the order of frequency, which were correlated with the RESULTS of Western blot. Subsequent investigations were focused on the physiologic role of NO in the reflex changes in bladder outlet activity during micturition in the rat. During IBC, the urethra exhibited reflex responses characterized by a decrease in RUR in conjunction with a rise in IBC. Administration of NOS inhibitor, Nw-nitro-L-arginine, reversibly decreased the magnitude and duration of RUR, and this effect was reversed by administration of L-arginine. From these RESULTS, it is suggested that the neuronal form of constitutive NOS in the bladder outlet synthesizes NO by its catalytic action, which mediates relaxation of bladder outlet during micturition.
Animals ; Arginine ; Blotting, Western ; Cerebellum ; Muscle, Smooth ; NADP ; NADPH Dehydrogenase ; Nerve Fibers ; Neurons ; Nitric Oxide Synthase ; Nitric Oxide* ; Rats* ; Reflex ; Relaxation* ; Urethra ; Urinary Bladder ; Urination

Axotomy of the vagal motor neurons by cervical vagotomy induces NADPH diaphorase staining due to increased nitric oxide synthase expression in both the rat dorsal motor nucleus and nucleus ambiguous; furthermore, cerical vagotomy leads to cell death of the dorsal motor nucleus cells. Subdiaphragmatic vagotomy axotomizes the vagal motor cells further from the brainstem than cervical vagotomy, and cuts the fibers running only to the abdominal viscera. Here we report that subdiaphragmatic vagotomy is sufficient to induce NADPH diaphorase staining in the dorsal motor nucleus but does not induce staining in the nucleus ambiguus. Because the neurons of the dorsal motor nucleus do not undergo cell death after subdiaphragmatic vagotomy and are able to re-enervate the gut, the increased nitric oxide synthase expression after distal axotomy may be related more to regeneration than degeneration.
Animal ; Fourth Ventricle/physiology* ; Fourth Ventricle/enzymology* ; Fourth Ventricle/cytology ; Male ; Motor Neurons/enzymology ; NADPH Dehydrogenase/metabolism* ; Rats ; Rats, Sprague-Dawley ; Vagotomy/methods* ; Vagus Nerve/physiology*

The Amyloid β peptide (Aβ) is a main component of senile plaques in Alzheimer's disease. Currently, NADPH oxidase (NOX) and mitochondria are considered as primary sources of ROS induced by Aβ. However, the contribution of NOX and mitochondria to Aβ-induced ROS generation has not been well defined. To delineate the relative involvement of NOX and mitochondria in Aβ-induced ROS generation and neuronal death in mouse cortical cultures, we examined the effect of NOX inhibitors, apocynin and AEBSF, and the mitochondria-targeted antioxidants (MTAs), mitotempol and mitoquinone, on Aβ-induced ROS generation and neuronal deaths. Cell death was assessed by measuring lactate dehydrogenase efflux in bathing media at 24 and 48 hrs after exposure to Aβ₁₋₄₂. Aβ₁₋₄₂ induced dose- and time-dependent neuronal deaths in cortical cultures. Treatment with 20 µM Aβ₁₋₄₂ markedly and continuously increased not only the DHE fluorescence (intracellular ROS signal), but also the DHR123 fluorescence (mitochondrial ROS signal) up to 8 hrs. Treatment with apocynin or AEBSF selectively suppressed the increase in DHE fluorescence, while treatment with mitotempol selectively suppressed the increase in DHR123 fluorescence. Each treatment with apocynin, AEBSF, mitotempol or mitoquinone significantly attenuated the Aβ₁₋₄₂-induced neuronal deaths. However, any combined treatment with apocynin/AEBSF and mitotempol/mitoquinone failed to show additive effects. These findings indicate that 20 µM Aβ₁₋₄₂ induces oxidative neuronal death via inducing mitochondrial ROS as well as NOX activation in mixed cortical cultures, but combined suppression of intracellular and mitochondrial ROS generation fail to show any additive neuroprotective effects against Aβ neurotoxicity.
Alzheimer Disease ; Amyloid beta-Peptides ; Amyloid* ; Animals ; Antioxidants* ; Baths ; Cell Death ; Fluorescence ; L-Lactate Dehydrogenase ; Mice* ; Mitochondria ; NADP* ; NADPH Oxidase* ; Neurons* ; Neuroprotective Agents ; Oxidative Stress ; Plaque, Amyloid