World J Mens Health.  2017 Dec;35(3):196-204. 10.5534/wjmh.17023.

Penile Erection Induced by Scoparone from Artemisia capillaris through the Nitric Oxide-Cyclic Guanosine Monophosphate Signaling Pathway

• Choi BR
• Kim HK
• Park JK

• KMID: 2398569
• DOI: http://doi.org/10.5534/wjmh.17023

Abstract

PURPOSE
The objective of this study was to evaluate the relaxant effect of scoparone from Artemisia capillaris on rabbit penile corpus cavernosum smooth muscle (PCCSM) and to elucidate the mechanism of action of scoparone for the treatment of erectile dysfunction (ED).
MATERIALS AND METHODS
PCCSM that had been precontracted with phenylephrine was treated with 3 Artemisia herbs (A. princeps, A. capillaris, and A. iwayomogi) and 3 fractions (n-hexane, ethyl acetate, and n-butanol) with different concentrations (0.1, 0.5, 1.0, and 2.0 mg/mL). Four components (esculetin, scopoletin, capillarisin, and scoparone) isolated from A. capillaris were also evaluated. The PCCSM was preincubated with Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME) and 1H-[1,2,4] oxadiazolo [4,3-a]quinoxalin-1-one (ODQ). Cyclic nucleotides in the perfusate were measured by a radioimmunoassay. The interactions of scoparone with udenafil and rolipram were also evaluated.
RESULTS
A. capillaris extract relaxed PCCSM in a concentration-dependent manner. Scoparone had the highest relaxant effect on PCCSM among the 4 components (esculetin, scopoletin, capillarisin, and scoparone) isolated from the ethyl acetate fraction. The application of scoparone on PCCSM pretreated with L-NAME and ODQ led to significantly less relaxation. Scoparone also increased the cyclic guanosine monophosphate (cGMP) levels in the perfusate in a concentration-dependent manner. Furthermore, scoparone enhanced udenafil- and rolipram-induced relaxation of the PCCSM.
CONCLUSIONS
Scoparone relaxed the PCCSM mainly by activating the nitric oxide-cGMP signaling pathway, and it may be a new promising treatment for ED patients who do not completely respond to udenafil.

Keyword

Coumarins; Erectile dysfunction; Nitric oxide; Phosphodiesterase 5 inhibitors

MeSH Terms

Artemisia*
Coumarins
Erectile Dysfunction
Guanosine Monophosphate*
Guanosine*
Humans
Male
Muscle, Smooth
NG-Nitroarginine Methyl Ester
Nitric Oxide
Nucleotides, Cyclic
Penile Erection*
Phenylephrine
Phosphodiesterase 5 Inhibitors
Radioimmunoassay
Relaxation
Rolipram
Scopoletin
Coumarins
Guanosine
Guanosine Monophosphate
NG-Nitroarginine Methyl Ester
Nitric Oxide
Nucleotides, Cyclic
Phenylephrine
Phosphodiesterase 5 Inhibitors
Rolipram
Scopoletin

Figure

• 

  Fig. 1 Total ion chromatograms of the ethyl acetate extract of Artemisia capillaris and purified scoparone. Crude ethyl acetate extract of A. capillaris extract (A), purified scoparone (B), ultraviolet spectrum of scoparone at 34.09 minutes from the crude extract photodiode array chromatogram (C), and a mass spectrogram of scoparone at 34.09 minutes from the crude extract total ion chromatogram (D).

• 

  Fig. 2 Percentage of relaxation induced by the ethanol extract of Artemisia herbs (n=4). The penile corpus cavernosum smooth muscle that had been precontracted with 10−5 M of phenylephrine (Phe) was treated with 4 concentrations of Artemisia herbs. The submaximal penile contractile responses induced by the Phe solution were taken as 100%, and all subsequent responses to Artemisia herbs were expressed as a percentage of this value. Each point represents the mean±standard deviation of the percentages (*p<0.05 vs. 0.1 mg/mL, **p<0.01 vs. 0.1 mg/mL).

• 

  Fig. 3 Percentage of relaxation induced by the fractions partitioned with n-hexane, ethyl acetate, and n-butanol (n=4). The penile corpus cavernosum smooth muscle that had been precontracted with 10−5 M of phenylephrine (Phe) was treated with 4 concentrations of each fraction. The submaximal penile contractile responses induced by Phe were taken as 100%, and all subsequent responses to Artemisia capillaris fractions were expressed as a percentage of this value. Each point represents the mean±standard deviation of the percentages (**p<0.01 vs. 0.1 mg/mL).

• 

  Fig. 4 Percentage of relaxation induced by esculetin, scopoletin, capillarisin, and scoparone (n=4). The penile corpus cavernosum smooth muscle that had been precontracted with 10−5 M of phenylephrine (Phe) was treated at 4 concentrations of esculetin, scopoletin, capillarisin, and scoparone. The submaximal penile contractile responses induced by Phe were taken as 100%, and all subsequent responses to esculetin, scopoletin, capillarisin, and scoparone were expressed as a percentage of this value. Each point represents the mean±standard deviation of the percentages. Statistical analysis was carried out by analysis of variance followed by the Bonferroni test (*p<0.05 vs. 10−7 M, **p<0.01 vs. 10−7 M).

• 

  Fig. 5 Relaxant effect of scoparone in L-phenylephrine (Phe)-induced contraction (n=4). Penile corpus cavernosum smooth muscle contracted by Phe (10−5 M) that was preincubated with Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME, 10−3 M) (A) or 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, 10−5 M) (B) was treated with 4 concentrations of scoparone (10−7, 10−6, 10−5, and 10−4 M). The submaximal penile contractile responses induced by Phe were taken as the 100% values, and all subsequent responses to scoparone were expressed as a percentage of this value. Each point represents the mean ± standard deviation of the percentages. Statistical analysis was carried out by analysis of variance followed by the Bonferroni test (*p<0.05 vs. scoparone, **p<0.01 vs. scoparone).

• 

  Fig. 6 Effects of scoparone (10−7, 10−6, 10−5, and 10−4 M) on cyclic guanosine monophosphate (cGMP) levels in the perfusate. Each point represents the mean±standard deviation of the percentages. Statistical analysis was carried out by analysis of variance followed by the Bonferroni test (**p<0.01 vs. control).

• 

  Fig. 7 Interaction of scoparone (SCOP, 10−6 M) with udenafil (UDE, 10−7 M) or rolipram (ROL 10−6 M) (n=4). Effects of different concentrations of UDE (A) and ROL (B) in penile corpus cavernosum smooth muscle (PCCSM) contracted by phenylephrine (Phe, 10−5 M). Each point represents the mean ± standard deviation of the percentages of maximal relaxation of the preceding submaximal contractile responses. Statistical analysis was carried out by analysis of variance followed by the Bonferroni test (**p<0.01 vs. UDE or ROL). (A) UDE+SCOP refers to scoparone-induced relaxation in the udenafil-preincubated PCCSM. (A) SCOP+UDE indicates udenafil-induced relaxation in the scoparone-preincubated PCCSM. (B) ROL+SCOP refers to scoparone-induced relaxation in the rolipram-preincubated PCCSM. (B) SCOP+ROL indicates rolipram-induced relaxation in the scoparone-preincubated PCCSM.

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