Nguyen Thi Thu Huong

Crude saponin extracted from Vietnamese ginseng and its major constituent majonoside-R2 attenuate the psychological stress- and foot-shock stress-induced antinociception in mice.

Effects of Vietnamese ginseng (VG) crude saponin and majonoside-R2, a major saponin constituent, on the psychological stress- and foot shock stress-induced antinociception in the tail pinch test were examined in mice. VG crude saponin (6.2, 12.5, and 25 mg/kg, P.O.) attenuated psychological stress- but not foot shock stress-induced antinociceptive response, whereas majonoside-R2 (3, 6.2, and 12.5 mg/kg, P.O. and i.p.), as well as naloxone (2 mg/kg, i.p.), suppressed both psychological stress- and foot shock stress-induced antinociception. Pretreatment with the crude saponin (12.5 mg/kg, P.O.) or majonoside-R2 (6.2 mg/kg, P.O.) for 5 days followed by the treatment in combination with stress for next 5 days did not affect the development of adaptation to foot shock stress, but they significantly suppressed the antinociceptive action of the stress measured on the first, second, and third day during the stress exposure period. Majonoside-R2 (6.2 mg/kg, P.O.) but not the crude saponin (12.5 mg/kg, P.O.) significantly blocked the development of adaptation to psychological stress. These results suggest that VG crude saponin has the suppressing effect on psychological stress- and foot shock stress-induced antinociception and that majonoside-R2 is important for the action of the saponin.

Majonoside-R2 reverses social isolation stress-induced decrease in pentobarbital sleep in mice: possible involvement of neuroactive steroids.

Majonoside-R2 (MR2) is a major ocotillol-type saponin constituent of Vietnamese ginseng. We investigated the effect of MR2 on the social isolation stress-induced decrease in pentobarbital sleep in mice, and elucidated the possible involvement of neurosteroidal sites of the GABA(A) receptor complex in the pharmacological activity of MR2. MR2 (3.1-6.2 mg/kg, i.p. or 5-10 microg, i.c.v.) dose-dependently reversed the decrease in pentobarbital sleep caused by social isolation stress to the level of sleep in the group-housed mice, but it had no effect on pentobarbital sleep in group-housed mice. Allotetrahydrodeoxycorticosterone (5alpha-pregnane-3alpha,21-diol-20-one, allo-THDOC; 12.5 microg, i.c.v.), the positive allosteric modulator of the GABA(A) receptor, and alpha-helical CRF(9-41) (alpha hCRF; 25 microg, i.c.v.), the corticotropin-releasing factor (CRF) antagonist, also reversed the decrease in pentobarbital sleep caused by social isolation stress. The reversing effects of i.c.v. MR2 and i.c.v. allo-THDOC on the decrease in pentobarbital sleep in isolated mice were significantly attenuated by pregnenolone sulfate (10 microg, i.c.v.), the steroidal negative allosteric modulator of the GABA(A) receptor. In contrast, when injected i.c.v., MR2, as well as allo-THDOC and alpha hCRF, significantly reversed the decrease in pentobarbital sleep induced by pregnenolone sulfate (10 microg, i.c.v.) and CRF (10 microg, i.c.v.) in group-housed mice. These results suggest that the reversing effect of MR2 on the social isolation stress-induced decrease in pentobarbital sleep is mediated by the neurosteroid site on the GABA(A) receptor complex in mice.

Majonoside-R2, a major constituent of Vietnamese ginseng, attenuates opioid-induced antinociception

The effects of majonoside-R2 on antinociceptive responses caused by the mu-opioid receptor agonist morphine and the selective kappa-opioid receptor agonist U-50, 488H were examined by the tail-pinch test in mice. Intraperitoneal (IP) or intracerebroventricular (ICV) injection of majonoside-R2 (3.1-6.2 mg/kg, IP or 5-10 micrograms/mouse, ICV) and diazepam (0.1-0.5 mg/kg, IP or 0.5-1.0 microgram/mouse, ICV), as well as an opioid receptor antagonist naloxone (2 mg/kg, IP or 5 micrograms/mouse, ICV), dose-dependently attenuated the antinociception caused by subcutaneously administered morphine and U-50,488H. Moreover, when co-administered ICV or intrathecally (IT) with morphine (4 micrograms/mouse) or U-50,488H (60 micrograms/mouse), majonoside-R2 (5-20 micrograms/mouse) also exhibited antagonism against the antinociceptive action of these opioid receptor agonists in the tail-pinch test. The inhibitory effects of majonoside-R2 (10 micrograms/mouse, ICV) and diazepam (1 microgram/mouse, ICV) were reversed by flumazenil (2.5 micrograms/mouse, ICV), a selective benzodiazepine receptor antagonist, and picrotoxin (0.25 microgram/mouse, ICV), a GABA-gated chloride channel blocker. These results suggest that majonoside-R2 attenuates the opioid-induced antinociception by acting at the spinal and supraspinal levels, and that the GABAA receptor complex at the supraspinal level is involved in the effect of ICV administered majonoside-R2.

Effects of majonoside-R2 on pentobarbital sleep and gastric lesion in psychologically stressed mice

The effects of Vietnamese ginseng (VG) and its major constituent majonoside-R2 on pentobarbital-induced sleep and gastric lesion in psychologically stressed mice were examined. Psychological stress exposure for 30 min significantly decreased the duration of pentobarbital (50 mg/kg, IP)-induced sleep in mice. VG extract (50 mg/kg, PO), VG saponin (25 mg/kg, PO), and majonoside-R2 (3.1-12.5 mg/kg, PO and IP) had no effect on pentobarbital sleep in unstressed control mice, but these drugs significantly recovered pentobarbital sleep decreased by psychological stress to the level of unstressed control animals. On the other hand, Panax ginseng (PG) extract (50-100 mg/kg, PO) failed to affect pentobarbital sleep in psychologically stressed mice. The effect of majonoside-R2 on psychological stress-induced decrease in the hypnotic activity of pentobarbital was significantly blocked by flumazenil (1 mg/kg, IV), a selective benzodiazepine antagonist. Diazepam (0.1 mg/kg, IP) significantly prolonged pentobarbital sleep in unstressed and psychologically stressed groups, and the effect of diazepam was significantly attenuated by the same dose of flumazenil. Naloxone (0.5-5 mg/kg, IP), an opioid antagonist, had no effect on pentobarbital sleep in unstressed or psychologically stressed animals. Psychological stress exposure for 16 h caused gastric lesion in mice. VG extract (25-50 mg/kg, PO) and majonoside-R2 (6.2-12.5 mg/kg, PO), as well as diazepam and naloxone, produced the protective action on gastric lesion in psychologically stressed mice. These results suggest that VG and its major constituent majonoside-R2 have the protective effects on the psychological stress-induced pathophysiological changes and that benzodiazepine receptors are partly implicated in the effects of majonoside-R2.

Suppressive Effects of Vietnamese Ginseng Saponin and Its Major Component Majonoside-R2 on Psychological Stress-Induced Enhancement of Lipid Peroxidation in the Mouse Brain

We investigated the in vivo effects of Vietnamese ginseng saponin (VG saponin) and its major component majonoside-R2 (MR2) on psychological stress-induced enhancement of lipid peroxidation in the mouse brain. Psychological stress exposure using a communication box system for 4 h significantly increased the content of thiobarbituric acid reactive substance (TBARS), an index of lipid peroxidation activity, in the brain. Pretreatment with VG saponin (15-25 mg/kg, PO) and MR2 (1-10 mg/kg, IP) significantly attenuated the psychological stress-induced increase in TBARS content in the brain. The aglycone of MR2 (MR2-aglycone: 1.2 mg/kg, IP), at the equivalent dose of MR2 (i.e., 3 mg/kg, IP), also produced the suppressive effect on the increase in the TBARS content. The in vivo suppressive effect of MR2 was dose dependently attenuated by flumazenil (3 and 10 mg/kg, IP), a benzodiazepine receptor antagonist, and pregnenolone sulfate (10 mg/kg, IP), a neurosteroidal negative allosteric modulator of GABA(A) receptors. These findings suggest that VG saponin and its major component MR2 have preventive effects on the psychological stress-induced brain cell membrane damage, and that the effect of MR2 is partly due to enhancement of GABA(A)-ergic systems in the brain.

INVOLVEMENT OF SUPRASPINAL GABA RECEPTORS IN MAJONOSIDE-R2 SUPPRESSION OF CLONIDINE-INDUCED ANTINOCICEPTION IN MICE

Majonoside-R2 (MR2) is a major constituent of Vietnamese ginseng (Panax vietnamensis, Ha et Grushv. Araliaceae) that is known to exhibit antagonistic activity against opioid-induced antinociception. In this study, we investigated the effect of MR2 on the antinociception caused in mice by the alpha2-adrenoceptor agonist clonidine, and elucidated the role of supraspinal GABAergic systems in this effect of MR2. The systemic administration of clonidine (0.5-2.5 mg/kg, s.c.) dose-dependently suppressed the nociceptive response of mice in the tail-pinch and hot-plate tests. The intraperitoneal (i.p.), intracerebroventricular (i.c.v.) or intrathecal (i.t.) administration of idazoxan (a selective alpha2-adrenoceptor antagonist) significantly antagonized the antinociceptive effect of clonidine in both tests. MR2 administered systemically (1.5-6.2 mg/kg, i.p.) or centrally (5-10 microg/mouse, i.c.v. or i.t.) dose-dependently antagonized the clonidine (1 mg/kg, s.c.)-induced antinociception in the tail-pinch test but not in the hot-plate test. The antagonistic effect of i.c.v. MR2 on the systemic clonidine-induced antinociception in the tail-pinch test was significantly reversed by i.c.v. administrations of the selective benzodiazepine receptor antagonist flumazenil (5 microg/mouse) and the GABA(A) antagonist picrotoxin (0.25 microg/mouse). These results suggest that the supraspinal GABA(A)/benzodiazepine receptors are involved in the antagonistic effect of MR2 on the clonidine-induced antinociception in the tail-pinch test in mice.

INVOLVEMENT OF SUPRASPINAL GABA-ERGIC SYSTEMS IN CLONIDINE-INDUCED ANTINOCICEPTION IN THE TAIL-PINCH TEST IN MICE

We investigated the involvement of supraspinal GABAergic systems in the antinociceptive effect of clonidine using the tail-pinch test in mice. Muscimol (31.2-250 ng/mouse, i.c.v.) and R(+)-baclofen (10-100 ng/mouse, i.c.v.), selective agonists for the GABA(A) and GABA(B) receptors, respectively, significantly attenuated the antinociceptive effect of subcutaneously (s.c.) administered clonidine (1 mg/kg) in a dose-dependent manner. The attenuating effect of muscimol (62.5 ng/mouse, i.c.v.) on the clonidine-induced antinociception was significantly blocked by the GABA(A) antagonists bicuculline (100-400 ng/mouse, i.c.v.) and picrotoxin (250 ng/mouse, i.c.v.) but not by the GABA(B) antagonist 2-hydroxysaclofen (10 microg/mouse, i.c.v.). On the other hand, the attenuating effect of R(+)-baclofen (50 ng/mouse, i.c.v.) was blocked by the coadministration with 2-hydroxysaclofen (20 microg/mouse), but was not affected by the coadministration with bicuculline (400 ng/mouse). These results indicate that both supraspinal GABA(A) and GABA(B) receptors play inhibitory roles in the antinociception caused by systemically administered clonidine.

Effects of vietnamese ginseng on opioid agonist — and conditioned fear stress-induced antinociception

We investigated the effects of Vietnamese ginseng (VG) extract, VG saponin and the VG major saponin constituent majonoside-R2 on opioid receptor agonist-induced antinociception using the tail-pinch and hot-plate tests in mice and on conditioned fear stress-induced antinociception using the tail-flick test in rats. VG extract (50-100 mg/kg, p.o.), VG saponin (12.5-25 mg/kg, p.o.) and majonoside-R2 (6.2-12.5 mg/kg, p.o.), as well as Panax ginseng extract (PG extract, 50-100 mg/kg, p.o.), dose-dependently attenuated the μ-opioid agonist morphine-induced antinociception in the tail-pinch and hot-plate tests. Moreover, repeated administration of VG saponin and majonoside-R2 suppressed the development of morphine tolerance in the tail-pinch test. VG extract (100-200 mg/kg, p.o.) also dose-dependently blocked the antinociceptive effects of the selective κ-opioid agonist U-50, 488H in the tail-pinch and hot-plate tests, while PG extract (100-200 mg/kg, p.o.) dose-dependently attenuated the U-50,488H-induced antinociception in the hot-plate test but not in the tail-pinch test. VG saponin (6.2-25 mg/kg, p.o.) blocked the U-50,488H-induced antinociception in the tail-pinch test but not in the hot-plate test. Furthermore, VG saponin (25 mg/kg, i.p.) and majonoside-R2 (6.2 mg/kg, i.p.), as well as naloxone (2 mg/kg, i.p.), reversed the tail-flick latency increased by conditioned fear stress in rats. These results indicate that VG and its major saponin constituent, majonoside-R2, attenuate the antinociception caused by opioid agonists and conditioned fear stress.

Effect of Vietnamese Ginseng on the phagocytosis in vitro and in vivo.

The effects of Vietnamese ginseng crude extract (VG extract), total saponin (VG saponin) and its major saponin component, majonoside-R2, on phagocytosis were examined in mice by bactericidal and carbon clearance tests. Escherichia coli (E. coli) ATCC 25922 was used to induce the acute toxicity and activate the phagocytic activity of phagocytes in both in vitro and in vivo bactericidal tests. Pretreatment with VG extract (500 mg/kg, oral administration, p.o.) and majonoside-R2 (50 mg/kg, intraperitoneal administration, i.p.) protected the animals from the acute toxicity of E. coli ATCC 25922 and significantly increased the phagocytic index in both in vitro and in vivo bactericidal tests. Moreover, VG extract (100-500 mg/kg, p.o.), VG saponin (25 mg/kg, i.p.) and majonoside-R2 (10 mg/kg, i.p.), as well as zymosan A, a non-specific phagocytic stimulant, also increased the phagocytic index evaluated by the carbon clearance test. These results indicate that Vietnamese ginseng enhances the phagocytic activity of phagocytes, and suggest that majonoside-R2 plays an important role in this effect.