• The Korea Journal of Herbology
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• v.30 no.3
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• pp.55-61
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• 2015

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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.28 no.2
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• pp.169-177
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• 2014

This study aimed to investigate the relaxation effects and its underlying mechanisms of Epimedium koreanum Nakai(EK) in phenylephrine(PE) treated isolated rabbit corpus cavernosum smooth muscle. The dose-dependent relaxation responses of phenylephrine(PE, $1{\times}10^{-6}M$)-precontracted strips to EK at $0.01-3.0mg/m{\ell}$ were measured and also observed after endothelial denudation using organ bath. To analyze the underlying mechanisms of EK-induced relaxation, $N{\omega}$-nitro-L-arginine(L-NNA), methylene blue(MB), tetraethylammonium chloride(TEA), indomethacin(IM) were pretreated before EK extract infused into precontracted strips induced by PE. To investigate cytotoxic activity and nitric oxide(NO) concentration of EK extract on EA.hy926 cells, mitochondrial dehydrogenase activity(MTT) assay and nitric oxide detection kit were used. The cavernous strips were significantly relaxed by EK extract at $0.3mg/m{\ell}$, $1.0mg/m{\ell}$, $3.0mg/m{\ell}$ and the relaxation responses of PE-precontracted strips denuded endothelium also inhibited in comparison with intact endothelium. The pretreatment of L-NNA, MB, TEA reduced EK extract-induced endothelium-dependent relaxation, but the pretreatment of IM didn't affect EK extract-induced endothelium-dependent relaxation. When EK extract was applicated on EA.hy926 cells, the NO concentration was increased. Our findings have shown that EK extract exerts a relaxing effect on corpus cavernosum in part by suppressing influx of extracellular $Ca^{2+}$ through activating 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.

• Biomolecules & Therapeutics
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• v.2 no.2
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• pp.136-140
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• 1994

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.

• Biomolecules & Therapeutics
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• v.11 no.4
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• pp.232-237
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• 2003

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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.29 no.2
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• pp.180-188
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• 2015

This study was conducted to investigate the relaxation effects and its mechanisms of Nelumbinis Semen(NS) extract in isolated rabbit corpus cavernous tissues. In order to examine the relaxation effects and its mechanisms of NS, we treated the ethanol extract of NS(0.01-3.0 mg/ml) and indomethacin(IM), tetraethylammonium chloride(TEA), Nω -nitro-L-arginine (L-NNA), methylene blue(MB) were treated before NS extract to contracted strips induced by PE 1 μM. We also treated calcium chloride(Ca) 1 mM after pretreatment of NS extract in Ca²⁺-free krebs-ringer solution to contracted strips induced by PE. Cell viability and NO concentration on human umbilical vein endothelial cell(HUVEC) was measured by MTT assay, Griess reagent system. eNOS production was investigated by histochemical and immunohistochemical staining. NS extract was significantly affected on the relaxation of cavernous strips and NS extract-induced relaxation was not different by pretreatment of IM, TEA, MB, but inhibited by the pretreatment of L-NNA. And increase of contraction induced by Ca²⁺ addition, in a Ca²⁺-free solution, was decreased by pretreatment of NS. NO concentration on HUVEC was increased. When NS extract was applicated on corpus cavernosum of penis(CCP) in SHR, ratio of smooth muscles to collage fibers by PE was decreased and formation of eNOS around helicine artery was increased. These results suggest that CCP relaxation effects of NS extract are shown by suppressing influx of extracellular Ca²⁺ through the production of NO and eNOS.

• The Journal of Internal Korean Medicine
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• v.37 no.4
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• pp.591-600
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• 2016

Objective: This study aimed to examine the relaxation effects and underlying mechanisms of Cynomorii herba (CH) extract in isolated rabbit corpus cavernous tissues.Methods: We experimented with CH extract (0.01-3.0 mg/mL). Nω-nitro-L-arginine (L-NNA) was experimented before the CH extract to contracted strips induced by phenylephrine (PE, 1 μM)and compared with nonexperimented. In addition, we experimented with calcium chloride (Ca²⁺, 1 mM) after pretreatment of the CH extract in Ca²⁺-free Krebs-Ringer solution to contracted strips induced by PE. The cell viability and nitric oxide (NO) concentration of human umbilical vein endothelial cells (HUVECs) were measured by an methylthiazol-2-yl-2, 5-diphenyl tetrazoliumbromide (MTT) assay and Griess reagent system. The ratio of smooth muscles to collagen fibers, in addition to eNOS- and PDE-5-positive reactions, was examined by histochemical and immunohistochemical staining.Results: The CH extract significantly induced the relaxation of the cavernous strips, and the pretreatment with L-NNA inhibited CH extract-induced relaxation. The L-NNA pretreatment reduced the increased contraction induced by the addition of Ca²⁺in Ca²⁺-free solution. Furthermore, the NO concentration of the HUVECs increased. When the CH extract was applied to the corpus cavernosum of the penis (CCP) of Sprague Dawley rats, the ratio of smooth muscles to collagen fibers by PE and the formation of eNOS around the helicine artery increased. However, the CH extract treatment decreased PDE-5 positive reactions.Conclusions: These results show that the relaxation effects induced by the CH extract are associated with the suppression of the influx of extracellular Ca²⁺ via the production of NO and eNOS and inhibition of PDE-5.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.4
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• pp.980-990
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• 2003

The effect of Gamigyuibi-tang on the penile erection induced by apomorphine HCI and on the erectile dysfunction induced by p,p-DDE, an environmental hormone derivate of DOT, were monitored using male cats. The changes of penile length, diameter, erectic periods and histological profiles of corpus cavernosum and corpus spongiosum were observed with blood testosterone levels. In conclusion, dose-dependent and significant increase of penile length, diameter, erectic periods and blood testosterone levels were detected in the Gamigyuibitang-dosing groups compared to other groups. In addition, it is also demonstrated that the increasement of congestion of blood vessels and dilation of connective tissues, and decreasement of adipocytes in the corpus cavernosum and/or corpus spongiosum of the Gamigyuibitang-dosing groups. According to these results, it is considered that Gamigyuibitang has some augmentation effect against to apomorphine HCI inducing penile erection and it also suggested that Gamigyuibitang has favorable effect to treatment of erectic dysfunctions induced by p,p-DDE.

• Proceedings of the PSK Conference
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• 2001.10a
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• pp.145.1-145.1
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• 2001