Nguyen, Hoai Bac;Lee, Shin Young;Park, Soo Hyun;Han, Jun Hyun;Lee, Moo Yeol;Myung, Soon Chul
The Korean Journal of Physiology and Pharmacology
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v.19
no.3
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pp.257-262
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2015
It is well known that cigarette smoke can cause erectile dysfunction by affecting the penile vascular system. However, the exact effects of nicotine on the corpus cavernosum remains poorly understood. Nicotine has been reported to cause relaxation of the corpus cavernosum; it has also been reported to cause both contraction and relaxation. Therefore, high concentrations of nicotine were studied in strips from the rabbit corpus cavernosum to better understand its effects. The proximal penile corpus cavernosal strips from male rabbits weighing approximately 4 kg were used in organ bath studies. Nicotine in high concentrations ($10^{-5}{\sim}10^{-4}M$) produced dose-dependent contractions of the corpus cavernosal strips. The incubation with $10^{-5}M$ hexamethonium (nicotinic receptor antagonist) significantly inhibited the magnitude of the nicotine associated contractions. The nicotine-induced contractions were not only significantly inhibited by pretreatment with $10^{-5}M$ indomethacin (nonspecific cyclooxygenase inhibitor) and with $10^{-6}M$ NS-398 (selective cyclooxygenase inhibitor), but also with $10^{-6}M$ Y-27632 (Rho kinase inhibitor). Ozagrel (thromboxane $A_2$ synthase inhibitor) and SQ-29548 (highly selective TP receptor antagonist) pretreatments significantly reduced the nicotine-induced contractile amplitude of the strips. High concentrations of nicotine caused contraction of isolated rabbit corpus cavernosal strips. This contraction appeared to be mediated by activation of nicotinic receptors. Rho-kinase and cyclooxygenase pathways, especially cyclooxygenase-2 and thromboxane $A_2$, might play a pivotal role in the mechanism associated with nicotine-induced contraction of the rabbit corpus cavernosum.
Journal of Physiology & Pathology in Korean Medicine
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v.29
no.6
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pp.485-491
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2015
This study was aimed to investigate the relaxation effects of Eucomiae Cortex (EC) extract in isolated rabbit corpus cavernosum smooth muscle and its mechanism. To evaluate the relaxation of EC extract in rabbit corpus cavernosum, EC extract was treated in corporal strips which were precontracted with phenylephrine(PE). To study its mechanism, Nω-nitro-L-arginine (L-NNA) was pretreated after infuse of EC extract and compared with non-treated. In calcium chloride (Ca2+) -free krebs solution, EC extract and Ca2+ 1 mM were infused by turns after Ca2+ 1 mM was treated into corporal strips contracted by PE. Cell ability, nitric oxide (NO) and epithelial nitric oxide synthase (eNOS) on human umbilical vein endothelial cell (HUVEC) were measured by MTT assay, Griess reagent system and histochemical, immunohistochemical methods. EC extract showed a significant relaxation effects on the corporal strips, this effects were inhibited by pretreatment of L-NNA. EC extract inhibited the increase of contraction by Ca2+ influx in Ca2+-free krebs solution, and eNOS positive reaction in corpus cavernosum, NO production in HUVEC increased by treatment of EC extract. These result suggest that the relaxation effects of EC extract in isolated corpus cavernosum smooth muscle are involved in increase of eNOS and NO production, blocking of extracellular Ca2+ influx.
Park, Mi-Sun;Kim, Jin-Bo;Hong, Eun-Ju;Hong, Sung-Cheul
YAKHAK HOEJI
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v.41
no.3
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pp.370-380
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1997
The role of nitric oxide (NO) on the non-adrenergic non-cholinergic (NANC) relaxations induced by the short and prolonged electrical field stimulation (EFS) has been studied in the rabbit corpus cavernosum. In the presence of atropine and guanethidine the prolonged EFS (2-16 Hz) of corpus cavernosal strips precontracted with phenylephrine produced frequency-dependent relaxations, which were abolished by tetrodotoxin as shown in the relaxations induced gy the short EFS, indicating that their orgin is NANC nerve stimulation. $N^G$-nitro-L-arginine (L-NNA), inhibitor of nitirc oxide synthase, caused a concentration-dependent inhibition to the NANC relaxation, and at 100 M L-NNA the relaxation were virtually abolished. The inhibitory effect of L-NNA was reversed by L-arginine. Hemoglobin abolished the relaxations to NO and also caused a concentration-dependent inhibition of the NANC relaxation. The hemoglobin-resistant relaxation induced by EFS was eliminated by L-NNA. Methylene blue significantly reduced the NANC relaxation in a conentration-dependent manner. The NANC relaxation was not affected by a VIP-inactivating pepridase, alpha0chymotrypsin, whereas VIP-induced relaxation was completely abolished. NO- and VIP-induced relaxation were not affected by L-NNA. These results indicate that the NANC relaxation induced by prolonged EFS of the rabbit corpus cavernosum is mediated by NO-guanosine 3',5'-cyclic monophosphate pathway as shown in the relaxation induced by the short EFS, and that VIP release is not essential for the NANC relaxation of the rabbit corpus cavernosum and VIP is not involved the generation fo NO.
Journal of Physiology & Pathology in Korean Medicine
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v.27
no.5
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pp.602-610
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2013
This study was aimed to evaluate the cavernosal relaxation effect of Crataegii fructus(CF) in the contracted rabbit penile corpus cavernosum by agonists.In order to study the effect of CF on the vasoconstriction of rabbit penile corpus cavernosum, isolated rabbit penile corpus cavernosum tissues were used for the experiment using organ baths containing Krebs solution.To investigate the cavernosal relaxation of CF, CF extract at $0.01{\sim}3.0mg/m{\ell}$ was added after penile corpus cavernosum were contracted by norepinephrine(NE) $1{\mu}M$. To analyze the mechanism of CF's vasorelaxation, CF extract infused into contracted penile tissues by NE after each treatment of indomethacin(IM), $N{\omega}$-nitro-L-arginine(L-NNA), methylene blue(MB), tetraethylammonium chloride(TEA).To study the effect of CF on influx of extracellular calcium chloride($Ca^{2+}$) in penile tissues, in $Ca^{2+}$-free krebs solution, $Ca^{2+}$ 1 mM infused into contracted penile tissues by NE after pretreatment of CF. Cytotoxic activity of CF on human umbilical vein endothelial cell(HUVEC) was measured by MTT assay, and nitric oxide(NO) prodution was measured by Griess reagent. CF relaxed cavernosal strip with endothelium contracted by NE, but in the strips without endothelium, CF-induced relaxation was significantly inhibited. The pretreatment of L-NNA, MB, TEA decreased significantly on the cavernosal relaxation than not-treatment of them. But the pretreatment of IM had no significant effect on the cavernosal relaxation. In $Ca^{2+}$-free krebs solution, when $Ca^{2+}$ infused into contracted penile tissues by NE, pretreatment of CF inhibit contraction induced by adding $Ca^{2+}$.NO production wasn't increased by treatment of CF on HUVEC. This findings showed that CF is effective for the relaxation of rabbit penile corpus cavernosum, and we suggest that CF relax rabbit corpus cavernosal smooth muscle through multiple action mechanisms that include increasing the release of nitric oxide from corporal sinusoidal endothelium, inhibition of $Ca^{2+}$ mobilization into cytosol from the extracellular fluid, and maybe a hyperpolarizing action.
Objectives : This study aimed to investigate the effects of Shingi-whan(SG) on the male reproductive and sexual function, so we measured the spermatogenesis and the testicular toxicity in mice and the vasorelaxation in isolated rabbit corpus cavernosum smooth muscle. Methods : To evaluate effect on the spermatogenesis in mice, we prepared two groups, control group and SG group that was orally administered SG(1,000mg/kg) for 20 days, and compared. To analyze testicular toxicity in mice, we also prepared two groups, doxo group that was injected with doxorubicin (3mg/kg) on three times and doxo + SG group that was injected with doxorubicin and SG for 20 days, and compared. To investigate sexual function of SG in mice, we prepared three groups, normal group and aging elicited group consisting of 18-month-old mice, SG treated aging group that was orally administered SG for 60 days, and compared using histochemical staining on mice corpus cavernous tissues. In order to define the relaxation effects of SG, rabbit corpus cavernous tissues were prepared in $2{\times}2{\times}6mm$ sized strip. Then the dose-dependent relaxation responses of SG at 0.01-3.0 mg/ml in contracted strips induced by phenylephrine were measured. Results : The sperm density in dutus epididymis and the diameter of seminiferous tubules of SG group was significantly increased when compared to control group. The testicular weight and the diameter and height of epithelial layer of seminiferous tubules of doxo + SG group was significantly increased when compared to doxo group. The cavernous strips were significantly relaxed by SG extract In SG treated aging group, ratio of smooth muscles to collagen fibers and red blood cell count in venous sinus was increased as compared to aging elicited group. Conclusions : Our findings have shown that SG extract have effect on spermatogenesis and mitigating effect on doxo-induced testicular toxicity. Further, it also have the vasorelaxant effect on rabbit corpus cavernosum.
Proceedings of the Korean Society of Applied Pharmacology
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1996.04a
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pp.217-217
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1996
The putative role of vasoactive intestinal polypeptide (VIP) as non-adrenergic non-cholinergic (NANC) neurotransmitter has been studied in rabbit corpus cavernosum. In the presence of atropine and guanethidine the short and prolonged electrical field stimulation (EFS, 2~16 ㎐) induced a frequency-dependent relaxation which was abolished by tetrodotoxin (0.3 ${\mu}$M), a nerve conductance blocker. The neurogenic relaxant reponses were not affected in the presence of VIP-inactivating peptidase, ${\alpha}$-chymotrypsin (2 units/$m\ell$), whereas VIP-induced relaxation were completely abolished. Inhibition of nitric oxide synthase by N$\^$G/-nitro-L-arginine (10~100 ${\mu}$M) caused concentration-dependent inhibition to the neurogenic relaxant responses and at 100 ${\mu}$M the relaxations were virtually abolished. In contrast NO (3~30 ${\mu}$M) and VIP (0.001~l ${\mu}$M)-induced relaxation were unaffected. The inhibitory effect of L-NNA was reversed in the presence of L-arginine (5 mM), the precursor of the NO biosynthesis. Hemog1obin (20~60 ${\mu}$M), sequestering NO in the extracellular space, abolished the NO-evoked relaxation and also caused a concentration-dependent inhibition to the neurogenic relaxation. These observation indicate that NANC relaxation induced by prolonged EFS of rabbit corpus cavernosum is also mediated mainly by nitric oxide as same as that of short EFS, and suggest that VIP is not involved in NANC relaxation of rabbit corpus cavernosum and NO would not be produced by VIP in this tissue.
Objectives : These present study was designed to investigate the relaxation effects of Alpiniae Oxyphyllae Fructus(AOF) on isolated corpus cavernosum smooth muscle.Methods : Rabbit corpus cavernous tissues were prepared in strip. Then relaxation responses of AOF at 0.01-3 ㎎/㎖ in contracted strips induced by phenylephrine(PE) were measured. To evaluate mechanisms, indomethacin(IM) tetraethylammonium chloride(TEA), Nω-nitro-L-arginine(L-NNA), methylene blue(MB) were treated before AOF extract(0.1-3 ㎎/㎖) infused into precontracted strips induced by PE. And 1 mM Ca2+was infused into precontracted strips after pretreatment of AOF extract(3 ㎎/㎖) in Ca2+-free krebs-ringer solution. NO concentration was measured by Griess reagent system. Ratio of smooth muscles to collagen fibers and eNOS positive reaction were measured by histocheminal and immunohistochemical process.Results : The cavernous strips were significantly relaxed by AOF extract 0.1, 0.3, 1, 3 ㎎/㎖ and the pretreatment with IM 10 μM,L-NNA 100 μM, MB 10 μM inhibited relaxation of AOF compared to non-pretreatment, but the pretreatment with TEA 100 μM didn't affect relaxation of AOF. In a Ca2+-free solution, pretreatment with AOF reduced increase on contraction of strips by Ca2+supply than non-pretreatment. On HUVEC, NO concentration was increased. On corpus cavernosum of penis in Spontaneous Hypertensive Rat, ratio of smooth muscles to collagen fibers and eNOS positive reaction in AOF group were increased compared to PE groupConclusions : Taken this results, we can suggest that AOF extract exerts a relaxation effects on rabbit corpus cavernosum smooth muscle in part by suppressing influx of extracellular Ca2+throughout prostacyclin, the NO-cGMP system.
Journal of Physiology & Pathology in Korean Medicine
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v.27
no.6
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pp.809-817
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2013
The aim of this study was to evaluate the mechanism of vasodilation of Lespedezea cuneata(LC) in rabbit common carotid artery and cavernosal smooth muscle. LC relaxed arterial strips precontracted with norepinephrine and cavernosal strips precontracted with phenylephrine. The arterial relaxation effects of LC was endothelium-dependent. $N{\omega}$-nitro-L-arginine(L-NNA), NOS inhibitor, methylene blue(MB), cGMP inhibitor, indomethacin(IM), cyclo-oxygenase inhibitor and tetraethylammonium chloride(TEA), KCa-channel blocker attenuate the relaxation responses of LC in arterial strips. In $Ca^{2+}$-free krebs-ringer solution, pretreatment of LC extract significantly reduced the contraction induced by addition $Ca^{2+}$. L-NNA reduced LC extract-induced relaxation in cavernosal strips, but IM, TEA and MB didn't affect LC extract-induced relaxation. When LC extract was applicated on human umbilical vein endothelial cell, the nitric oxide concentration was increased. We conclude that in rabbit common carotid artery, LC may suppress influx of extra-cellular $Ca^{2+}$ through the release of endothelium derived relaxing factor including nitric oxide, prostacyclin, endothelium derived hyperpolarizing factor. And LC exerts a relaxing effect on corpus cavernosum through activating the NO.
To prove the hypothesis that NO- and N $O_2$-carrying molecules potentiate photorelaxation by generating NO, investigation was carried out using isolated rabbit and human corpus cavernosum. Corporal smooth muscle, in the presence or absence of endothelium, relaxed only slightly upon ultraviolet light (366 nm) irradiation. But, NO-and/or N $O_2$-containing compounds such as streptozotocin and $N^{G}$-nitro-L-arginine methyl ester significantly (p<0.01) enhanced photorelaxation in this tissue. In addition, $N^{G}$-nitro-D-arginine methyl ester, known to lack inhibitory action on NO synthase, showed concentration-dependent potentiation of the photorelaxation. Oxygen radical generating system via copper+ascorbic acid and guanylate cyclase inhibitor, methylene blue, significantly (p<0.05) inhibited the streptozotocin-potentiated photorelaxation. Nitrite was accumulated by photolysis of streptozotocin, $N^{G}$-nitro-L-arginine methyl ester and $N^{G}$-nitro-D-arginine methyl ester, in a concentration and exposure time dependent manner. These observations indicate that NO is a potent relaxant of rabbit and human corpus cavernosum and further support the hypothesis that NO is released by photolysis from NO- and N $O_2$-carrying molecules.lecules.
The present study examined functional effects of a new selective phosphodiesterase type 5 inhibitor, 1-[4-ethoxy-3-(6,7-dihydro-1-methyl-7-thioxo-3-propyl-1H-pyrazolo[ 4,3]pyrimidin-5-yl)phenylsulphonyl]-4-methyl piperazine (KJH-1002), in the isolated rabbit corpus cavernosum (RCC). Relaxing effects of KJH-1002 were also compared with those of sildenafil, which is currently used as an oral therapy for penile erectile dysfunction. In the isolated RCC precontracted with phenylephrine, both KJH-1002 and sildenafil in the concentration range of 1 to 1000 nM, produced a comparable potentiation of the electical field stimulation-induced relaxation in a concentration-dependent manner. In the sodium nitroprusside (SNP)-induced relaxation, the $IC_{50}$/ values, concentrations of SNP required to produce a 50% relaxation of the phenylephrine-induced contraction, were significantly decreased to the similar extent by treatments with KJH-1002 and sildenafil. The results suggest that a new selective phosphodiesterase type 5 inhibitor, KJH-1002, has an augmentative effect on penile erection comparable to that of sildenafil and can be useful for the treatment of erectile dysfunction.
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