Korean J Physiol Pharmacol.  2014 Oct;18(5):431-439. 10.4196/kjpp.2014.18.5.431.

Inhibitory Effects of Ginsenoside-Rb2 on Nicotinic Stimulation-Evoked Catecholamine Secretion

Affiliations
  • 1Department of Internal Medicine (Division of Pulmonary and Critical Care Medicine), Veterans Health Service Medical Center, Seoul 134-791, Korea.
  • 2Department of Anesthesiology and Pain Medicine, School of Medicine, Chosun University, Gwangju 501-759, Korea.
  • 3Department of Pharmacology, School of Medicine, Chosun University, Gwangju 501-759, Korea. dylim@chosun.ac.kr

Abstract

The aim of the present study was to investigate whether ginsenoside-Rb2 (Rb2) can affect the secretion of catecholamines (CA) in the perfused model of the rat adrenal medulla. Rb2 (3~30 microM), perfused into an adrenal vein for 90 min, inhibited ACh (5.32 mM)-evoked CA secretory response in a dose- and time-dependent fashion. Rb2 (10 microM) also time-dependently inhibited the CA secretion evoked by DMPP (100 microM, a selective neuronal nicotinic receptor agonist) and high K+ (56 mM, a direct membrane depolarizer). Rb2 itself did not affect basal CA secretion (data not shown). Also, in the presence of Rb2 (50 microg/mL), the secretory responses of CA evoked by veratridine (a selective Na+ channel activator (50 microM), Bay-K-8644 (an L-type dihydropyridine Ca2+ channel activator, 10 microM), and cyclopiazonic acid (a cytoplasmic Ca2+-ATPase inhibitor, 10 microM) were significantly reduced, respectively. Interestingly, in the simultaneous presence of Rb2 (10 microM) and L-NAME (an inhibitor of NO synthase, 30 microM), the inhibitory responses of Rb2 on ACh-evoked CA secretory response was considerably recovered to the extent of the corresponding control secretion compared with the inhibitory effect of Rb2-treatment alone. Practically, the level of NO released from adrenal medulla after the treatment of Rb2 (10 microM) was greatly elevated compared to the corresponding basal released level. Collectively, these results demonstrate that Rb2 inhibits the CA secretory responses evoked by nicotinic stimulation as well as by direct membrane-depolarization from the isolated perfused rat adrenal medulla. It seems that this inhibitory effect of Rb2 is mediated by inhibiting both the influx of Ca2+ and Na+ into the adrenomedullary chromaffin cells and also by suppressing the release of Ca2+ from the cytoplasmic calcium store, at least partly through the increased NO production due to the activation of nitric oxide synthase, which is relevant to neuronal nicotinic receptor blockade.

Keyword

Adrenal Medulla; Catecholamine secretion; Ginsenoside-Rb2 (Rb2); Nitric oxide synthase (NOS); NO production

MeSH Terms

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester
Adrenal Medulla
Animals
Calcium
Catecholamines
Chromaffin Cells
Cytoplasm
Dimethylphenylpiperazinium Iodide
Membranes
Neurons
NG-Nitroarginine Methyl Ester
Nitric Oxide Synthase
Rats
Receptors, Nicotinic
Veins
Veratridine
3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester
Calcium
Catecholamines
Dimethylphenylpiperazinium Iodide
NG-Nitroarginine Methyl Ester
Nitric Oxide Synthase
Receptors, Nicotinic
Veratridine

Figure

  • Fig. 1 Dose-dependent effects of ginsenoside-Rb2 (Rb2) on the secretory responses of catecholamines (CA) evoked by acetylcholine (ACh) from the perfused rat adrenal medulla. The CA secretion by a single injection of ACh (5.32×10-3 M) in a volume of 0.05 mL was evoked at 15 min intervals during loading with 3, 10 and 30 µM of Rb2 for 90 min as indicated by the arrow marks, respectively. The numbers in parentheses indicate the number of rat adrenal glands. Vertical bars on the columns represent the standard error of the mean (S.E.M.). Ordinate: the amounts of CA secreted from the adrenal gland (% of control). Abscissa: collection time of perfusate (min). Statistical difference was obtained by comparing the corresponding control with each concentration - treated group of Rb2. ACh-induced perfusate was collected for 4 minutes. *p<0.05, **p<0.01. ns, Not statistically significant.

  • Fig. 2 Time-course effects of Rb2 on the high K+-evoked CA secretory responses from the perfused rat adrenal medulla. The CA secretion by a single injection of K+ (5.6×10-2 M) in a volume of 0.05 mL was evoked at 15 min intervals during loading with 10 µM of Rb2 for 90 min as indicated by the arrow marks. high K+-induced perfusate was collected for 4 minutes. Other legends are the same as in Fig. 1. **p<0.01. ns, Not statistically significant.

  • Fig. 3 Time-course effects of Rb2 on the CA secretory responses evoked by from the perfused rat adrenal medulla. The CA secretion by perfusion of DPPP (10-4 M) for 2 min was induced at 20 min interval during loading with 10 µM of Rb2 for 90 min. DMPP-induced perfusate was collected for 8 minutes. Other legends are the same as in Fig. 1. **p<0.01. ns, Not statistically significant.

  • Fig. 4 Time-course effects of Rb2 on the CA secretion evoked by Bay-K-8644 from the perfused rat adrenal medulla. Bay-K-8644 (10-5 M) was perfused into an adrenal vein for 4 min at 15 min intervals during loading with Rb2 (10 µM) for 90 min. Other legends are the same as in Fig. 1. **p<0.01. ns, Not statistically significant.

  • Fig. 5 Time-course effects of Rb2 on the CA secretion evoked by cyclopiazonic acid from the perfused rat adrenal medulla. Cyclopiazonic acid (10-5 M) was perfused into an adrenal vein for 4 min at 15 min intervals during loading with Rb2 (10 µM) for 90 min. Other legends are the same as in Fig. 1. **p<0.01. ns, Not statistically significant.

  • Fig. 6 Time-course effects of Rb2 on veratridine-evoked CA secretion from the perfused rat adrenal medulla. Veratridine (5×10-5 M) was perfused into an adrenal vein for 4 min at 15 min intervals during loading with Rb2 (10 µM) for 90 min. Other legends are the same as in Fig. 1. **p<0.01.

  • Fig. 7 Effects of Rb2 plus L-NAME on the CA secretory responses evoked by acetylcholine from the perfused rat adrenal medulla. The CA secretion by a single injection of ACh (5.32×10-3 M) in a volume of 0.05 mL was evoked at 15 min intervals during simultaneous loading with Rb2 (10 µM) plus L-NAME (30 µM) for 90 min. Statistical difference was obtained by comparing the corresponding control (CONTROL) with Rb2-treated only group or group treated with Rb2+L-NAME. Other legends are the same as in Fig. 1. *p<0.05, **p<0.01. ns: Not statistically significant.

  • Fig. 8 Effects of Rb2 on nitric oxide (NO) production in the perfused rat adrenal medulla. Perfusate sample was taken for 8 min after loading the perfusion of Rb2 (10 µM) at a rate of 0.31 mL/min. Ordinate: the amounts of NO released from the adrenal medulla (% of control). Abscissa: Treatment (before and after Rb2). Statistical difference was made by comparing the control with Rb2-treated group. **p<0.01.

  • Fig. 9 Probable site of action of Rb2 at cholinergic nerve-chromaffin cell in the rat adrenal medulla.


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