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Ginsenosides induce differential antinociception and inhibit substance P induced-nociceptive response in mice.

Ginsenosides are main pharmacoactive molecules of ginseng. The antinociceptive activity of ginsenosides after intrathecal (i.t.) injection was examined in formalin test. We also investigated the effects of ginsenosides on substance P (SP) induced-pain behaviors by i.t. treatment using mice. Pretreatment of ginsenosides by i.t. induced the inhibition of biting and licking of hind paw injected with 1% formalin with dose-dependent manner. The ED50 was 23 (19-28, 95% C.I.) microg/mouse for acute phase and 15 (9-23, 95% C.I.) microg/mouse for tonic phase. Interestingly, cotreatment of ginsenosides with SP also inhibited SP-induced pain behaviors (scratching, licking or biting of hind portion of body) with dose-dependent manner. The ED50 for the inhibition of SP-induced pain behavior by ginsenosides was 30 (11-85, 95% C.I.) microg/mouse. These results suggest that ginsenosides have antinociceptive activity in formalin test and this effect is due to blocking of SP-induced nociceptive information to postsynaptic site(s) at the spinal level.

Analgesia↗

Can ginsenosides protect human erythrocytes against free-radical-induced hemolysis?

Many studies have focused on the free-radical-initiated peroxidation of membrane lipid, which is associated with a variety of pathological events. Panax ginseng is used in traditional Chinese medicine to enhance stamina and capacity to deal with fatigue and physical stress. Many reports have been devoted to the effects of ginsenosides, the major active components in P. ginseng, on the lipid metabolism, immune function and cardiovascular system. The results, however, are usually contradictory since the usage of mixture of ginsenosides cannot identify the function of every individual ginsenosides on the experimental system. On the other hand, every individual ginsenosides is not compared under the same experimental condition. These facts motivate us to evaluate the antioxidant effect of various individual ginsenosides on the experimental system of free-radical-initiated peroxidation: the hemolysis of human erythrocyte induced thermally by water-soluble initiator, 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH). The inhibitory concentration of 50% inhibition (IC(50)) of AAPH-induced hemolysis of the erythrocyte has been studied firstly and found that the order of IC(50) is Rb3 - Rb1< Rc>Re>Rh1>R1>Rg2>Rb3. Rg3, Rd and Rh2, however, act as synergistic prooxidants in the above experimental system. Rg1 does not show any synergistic antioxidative property. Although the antioxidative and prooxidative mechanism of various ginsenosides with or without TOH in AAPH-induced hemolysis of human erythrocytes will be further studied in detail, this information may be useful in the clinical usage of ginsenosides.

Amidines↗

Effects of population, age, and cultivation methods on ginsenoside content of wild American ginseng (Panax quinquefolium).

Genotype and environmental effects on ginsenoside content among eight wild populations of American ginseng (Panax quinquefolium) were investigated. Root concentrations of six ginsenosides were determined at the time of collection of plants from the wild (T0) and 2 years (T2) after transplanting roots from each of the eight populations to each of two different forest garden locations. Both location and population had significant effects on root and shoot growth. Overall, ginsenoside Rb1 was most abundant, followed by Rg1 and Re. Concentrations of Rg1 and Re were inversely related among and within populations. The relative ranking of populations differed depending upon the particular ginsenoside and sampling time. The relative importance of genotype and environment was not the same for all ginsenosides. Ginsenoside Re was influenced by population but not location, whereas Rb1, Rc, and Rb2 were influenced only by location (environment), while Rg1 and Rd were influenced by both. Ginsenoside levels were consistently lower, but growth was consistently higher at the more intensively managed garden location.

Ginsenosides↗

Simultaneous quantification of ginsenosides in American ginseng (Panax quinquefolium) root powder by visible/near-infrared reflectance spectroscopy.

Near-infrared reflectance spectroscopy (NIRS) was examined as a possible alternative to high-performance liquid chromatography (HPLC) for the analysis of ginsenosides from American ginseng (Panax quinquefolium) root powder (n = 26). NIR spectra were collected over 400-2500 nm. For each sample and individual ginsenoside quantified by HPLC, spectral data were regressed against the chemical data to develop prediction equations. The spectral prediction equations produced high correlation coefficient (1-VR) values and low standard errors of cross validation (SECV) values for the determination of individual and total ginsenosides. The contents of individual ginsenosides, Rb(1), Rb(2), Rc, Rd, Re, Rg(1), Ro, m-Rb(1), m-Rb(2), m-Rc, m-Rd, and total ginsenosides (X +/- SECV) were (1.29+/-0.18)%, (0.273+/-0.096)%, (0.298+/-0.052)%, (0.091+/-0.027)%, (1. 015+/-0.114)%, (0.116+/-0.018)%, (0.25+/-0.040)%, (0.776+/-0.116)%, (0.197+/-0.074)%, (0.239+/-0.083)%, (0.143+/-0.042)%, and (4.393+/-0.283)%, respectively. The (1-VR) values of cross validation were 0.877, 0.872, 0.955, 0.834, 0.899, 0.919, 0.325, 0.849, 0.902, 0.877, 0.871, and 0.963, respectively. Results indicated that the NIRS method could be used for the analysis of the major ginsenosides, Rb(1), Re, and m-Rb(1), as well as the total ginsenosides in American ginseng.

Calibration↗

Variable effects of American ginseng: a batch of American ginseng (Panax quinquefolius L.) with a depressed ginsenoside profile does not affect postprandial glycemia.

BACKGROUND: We have repeatedly reported that American ginseng (AG) with a specific ginsenoside profile significantly decreases postprandial glycemia. Whether this effect is reproducible using AG with a different profile is unknown. We therefore investigated the effect of a different batch of AG on glycemia following a 75 g oral glucose tolerance test (OGTT). METHODS: Using a randomized, single blind design, 12 normal subjects (six males and six females, aged 31+/-3 y, body mass index (BMI) 28+/-2 kg/m(2)) received 6 g AG or placebo 40 min before a 75 g OGTT. The protocol followed the guidelines for the OGTT, with venous blood samples drawn at -40, 0, 15, 30, 45, 60, 90 and 120 min. Ginsenosides in the AG were assessed by established methods for HPLC-UV. RESULTS: Repeated measures analysis of variance demonstrated that there was no significant effect of the AG on incremental plasma glucose (PG) or insulin (PI) or their areas under the curve Indices of insulin sensitivity (insulin sensitivity index (ISI)) and release (deltaPI(30-0)/deltaPG(30-0)) calculated from the OGTT were also unaffected. The AG contained 1.66% total ginsenosides, 0.90% (20S)-protopanaxadiol (PPD) ginsenosides, and 0.75% (20S)-protopanaxatriol (PPT) ginsenosides, with the following key ratios: PPD:PPT of 1.2, Rb(1):Rg(1) of 8.1, and Rb(2):Rc of 0.18. CONCLUSIONS: The present batch of AG was unable to reproduce the postprandial hypoglycemic effects we observed previously. Possible explanations for this discrepancy include marked decrements in total ginsenosides and the key ratios PPD:PPT, Rb(1):Rg(1), and Rb(2):Rc. These data suggest that the ginsenoside profile of AG might play a role in its hypoglycemic effects. The involvement of other components cannot, however, be precluded.

Adult↗

Ginsenoside F1 protects human HaCaT keratinocytes from ultraviolet-B-induced apoptosis by maintaining constant levels of Bcl-2.

Ginsenosides, the major active ingredients of ginseng, show a variety of biomedical efficacies such as antiaging and antioxidation. Here, we investigate the protective activity of the ginsenoside F1, an enzymatically modified derivative of ginsenoside Rg1, against ultraviolet-B-induced damage in human HaCaT keratinocytes. Ginsenoside F1 significantly reduced ultraviolet-B-induced cell death and protected HaCaT cells from apoptosis caused by ultraviolet B irradiation. Furthermore, ginsenoside F1 prevented ultraviolet-B-induced cleavage of poly(ADP-ribose) polymerase in HaCaT cells. In search of the molecular mechanism responsible for the antiapoptotic effect of ginsenoside F1, we find that protection from ultraviolet-B-induced apoptosis is tightly correlated with ginsenoside-F1-mediated inhibition of ultraviolet-B-induced downregulation of Bcl-2 and Brn-3a expression.

Apoptosis↗

Neurotrophic and neuroprotective actions of ginsenosides Rb(1) and Rg(1).

The ginsenosides have many pharmacological actions, including various actions on the nervous system. Our previous studies have demonstrated that two ginsenosides, Rb(1) and Rg(1) improve performance in a passive avoidance-learning paradigm and enhance cholinergic metabolism. The present study was designed to examine the cellular neurotrophic and neuroprotective actions of two pure ginsenosides in two model systems. PC12 cells were grown in the absence or presence of nerve growth factor (NGF) as a positive control, and different concentrations of Rb(1) or Rg(1). To assess neurotrophic properties, neurite outgrowth was quantified for representative fields of cells. After 8 days in culture, both ginsenosides enhanced neurite outgrowth in the presence of a sub-optimal dose of (2 ng/ml) NGF, but did not significantly stimulate neurite outgrowth in the absence of NGF. However, after 18 days in culture, both ginsenosides increased neurite outgrowth in the absence of NGF. SN-K-SH cells were grown in the absence or presence of MPTP or beta-amyloid to assess neuroprotection. Rb(1) and Rg(1) both reversed MPTP-induced cell death. beta-Amyloid-induced cell death was not reversed by either ginsenoside, but Rg(1) produced a modest enhancement of cell death in this model. These results suggest that these two ginsenosides have neurotrophic and selective neuroprotective actions that may contribute to the purported enhancement of cognitive function.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

Decreasing, null and increasing effects of eight popular types of ginseng on acute postprandial glycemic indices in healthy humans: the role of ginsenosides.

BACKGROUND: It is unclear whether other ginseng sources can replicate the glycemic-lowering efficacy observed previously with American ginseng and whether ginsenosides are mediators. We assessed the effect of eight popular ginseng types on postprandial plasma glucose (PG) and insulin (PI) indices, linking effects to ginsenoside profiles. METHODS: Using a double-blind, randomized, multiple-crossover design, 12 healthy participants (gender: 6M:6F, age: 34 +/- 3 y, BMI: 25.8 +/- 1.2 kg/m(2)) received 10 3g treatments: American, American-wild, Asian, Asian-red, Vietnamese-wild, Siberian, Japanese-rhizome, and Sanchi ginsengs and two placebos. Each treatment was given 40-minutes before a 75g-oral-glucose-tolerance-test (75g-OGTT) with blood drawn at -40, 0, 15, 30, 45, 60, 90, 120-minutes. HPLC-UV analysis quantified seven principal ginsenosides. RESULTS: Two-factor analysis showed the main effects of ginseng-type and time were significant for PG and PI, with an interaction for PG (p < 0.05). Subsequent one-factor analysis showed an effect of ginseng-type on 90-min-PG and 90-min-PI (p < 0.05). This was reflected in effects on peak-PG, area under the curve (AUC)-PG and AUC-PI (p < 0.05). But the effect on 90-min-PI and AUC-PI were significant (p < 0.05) only in overweight participants (BMI > 25 kg/m(2), n = 6). Planned comparisons with placebo showed a tendency for American ginseng and Vietnamese ginseng to lower 90-min-PG (p < 0.06), while Asian ginseng raised peak-PG and AUC-PG, American-wild ginseng raised 120-min-PG, and Siberian ginseng raised 90-min-PG, 120-min-PG, and AUC-PG (p < 0.05). Stepwise-multiple-regression assessed the protopanaxadiol:protopanaxatriol (PPD:PPT)-ginsenoside ratio as the sole predictor (p < 0.05) for 90-min-PG (beta = -0.43, r(2) = 0.072), AUC-PG (beta = -0.25, r(2) = 0.06), 90-min-PI (beta = -0.26, r(2) = 0.065), AUC-PI (beta = -0.20, r(2) = 0.04). CONCLUSIONS: Ginseng has variable glycemic effects, in which the PPD:PPT-ginsenoside ratio might be involved. But the low variance explained suggests the involvement of other unmeasured ginsenoside or non-ginsenoside components.

Acute Disease↗

Ginsenoside-Rh1 and Rh2 inhibit the induction of nitric oxide synthesis in murine peritoneal macrophages.

The effects of ginsenoside-Rh1 and Rh2 in the induction of nitric oxide (NO) synthesis by IFN-gamma plus LPS were investigated using murine peritoneal macrophages. The NO production from rIFN gamma plus LPS-treated macrophages was markedly reduced by ginsenoside-Rh1 or Rh2 in a dose dependent manner, but had no inhibitory effects by ginsenoside-Rb1, Rc or Re. In addition, treatment of the cells with ginsenoside-Rh2 6 hr before the stimulation with IFN-gamma plus LPS showed more inhibitory effect than the treatment with ginsenoside-Rh2 6 hr after or simultaneously with the stimulation with IFN-gamma plus LPS in the NO production. Ginsenoside-Rh2 also effectively inhibited IFN-gamma induced NO production when the cells were treated with IFN-gamma 6 hr after the treatment with ginsenoside-Rh2. Our findings suggest that this phenomenon might be caused by inhibition of priming signal such as IFN-gamma for the synergistic induction of NO synthesis.

Animals↗

Effects of ginsenosides on vasodilator nerve actions in the rat perfused mesentery are mediated by nitric oxide.

This study was designed to explore the effect of ginsenosides, saponins from Panax ginseng, on the vasodilator nerve actions in the rat perfused mesentery and the mechanism of this effect. In the rat perfusion mesentery, when adrenergic nerves were blocked by guanethidine (5 x 10(-6) M) and vascular muscle tone was increased with methoxamine (5 x 10(-6)-10(-5) M), transmural field stimulation produced a frequency-dependent vasodilator response, which is due to the release of calcitonin gene-related peptide; ginsenosides significantly suppressed this vasodilator response in a concentration-dependent manner (3-30 micrograms mL-1). After pretreatment with saponin (50 micrograms mL-1, 3 min) to damage endothelial cells, this suppressing effect of ginsenosides was unaltered. However, the effect was abolished by N omega-nitro-L-arginine methyl ester (L-NAME) (10(-4) M), an inhibitor of nitric oxide synthesis and addition of L-arginine (3 x 10(-4) M) restored this suppressing effect. Methylene blue (10(-5) M), an inhibitor of guanylate cyclase, also abolished the suppressing effect of ginsenosides. However, ginsenosides did not alter the relaxation responses caused by exogenous calcitonin gene-related peptide administration. We conclude that ginsenosides can produce an inhibitory effect on the vasodilator response prejunctionally in the rat perfused mesentery and that this effect of ginsenosides may be mediated by nitric oxide released from non-adrenergic, non-cholinergic nerves.

Adrenergic Agents↗

Molecular biological analysis of the effects of ginsenoside-Rb2 on albumin mRNA in streptozotocin-induced diabetic rats.

The mechanisms of the mRNA synthesis-promoting action of ginsenoside-Rb2, were investigated at the gene level. Rot analysis suggested that the previously reported increase in RNA polymerase activity as a result of administration of ginsenoside-Rb2 might be because of its effect on a specific gene. In this regard, albumin mRNA, which is expressed specifically in the liver, was assayed by northern blot hybridization using albumin cDNA in normal rats, diabetic control rats and diabetic rats given ginsenoside-Rb2. When the level of albumin mRNA in normal rats was set at 100, the level was reduced markedly to 32 in diabetic control rats. In contrast, in diabetic rats given ginsenoside-Rb2 the level was 0.54, significantly higher (69%) than that in diabetic rats given no ginsenoside-Rb2. In addition, poly(A)+RNA was purified from total RNA and subjected to hybridization, and poly(A)+RNA bands with different charges were measured by densitometry. The results of the measurement revealed changes dependent on the charge, and this was confirmed by autoradiography. We found no significant difference in the transcription activity of albumin mRNA, however, it showed only a tendency to increase. This suggests that ginsenoside-Rb2 has some effect on post-transcriptional regulation of the stability of mRNA itself. The results of Rot analysis suggest that ginsenoside-Rb2 affects a specific gene alone.

Albumins↗

Ginsenoside Rh2 reduces ischemic brain injury in rats.

Ginseng was incubated under mildly acidic conditions and its inhibitory effect on a rat ischemia-reperfusion model was investigated. When ginseng was treated with 0.1% hydrochloric acid at 60 degrees C, its protopanaxadiol saponins were transformed to diasteromeric ginsenoside Rg3 and Delta20-ginsenoside Rg3. When the transformed ginseng extract, of which the main component was ginsenosides Rg3, was treated with human intestinal microflora, the main metabolite was ginsenoside Rh2. Orally administered acid-treated ginseng (AG) extract and ginsenoside Rh2 potently protect ischemia-reperfusion brain injury. The ginsenoside Rh2 also inhibited prostaglandin-E2 synthesis in lipopolysaccharide-stimulated RAW264.7 cells, but showed no in vitro antioxidant activity. These results suggest that AG and ginsenoside Rh2 can improve ischemic brain injury.

Administration, Oral↗

Stereospecific effects of ginsenoside Rg3 epimers on swine coronary artery contractions.

Previous reports have shown that ginseng saponins, the active ingredients of Panax ginseng, induce relaxation of hormone- or high K(+)-induced blood vessel contraction. We recently demonstrated that 20(R)- and 20(S)-ginsenoside Rg(3) epimers regulate ion channel activities in a stereospecific manner. Here, we examined whether ginsenoside Rg(3) epimers also exhibit differential effects on swine coronary artery contractions induced by high K(+) or 5-HT. We found that treatment with 20(S)- but not 20(R)-ginsenoside Rg(3) caused a potent concentration-dependent, endothelium-independent relaxation of coronary artery contraction induced by 25 mM KCl. However, treatment with both 20(S)- and 20(R)-ginsenoside Rg(3) induced a significant, concentration-dependent relaxation of 3 microM 5-HT-induced coronary artery contractions in intact samples, while only 20(S)-ginsenoside Rg(3) inhibited coronary artery contraction in endothelium-denuded arteries. 20(S)- but not 20(R)-ginsenoside Rg(3) inhibited L-type Ca(2+) channel currents in a dose- and voltage-dependent manner. These results indicate that 20(S)- and 20(R)-ginsenoside Rg(3) epimers might exhibit stereospecific relaxation effects on swine coronary artery contractions caused by high K(+) and 5-HT receptor activation.

Animals↗

Enzymatic preparation of ginsenosides Rg2, Rh1, and F1.

During investigation of the hydrolysis of a protopanaxatriol-type saponin mixture by various glycoside hydrolases, crude preparations of beta-galactosidase from Aspergillus oryzae and lactase from Penicillium sp. were found to produce two minor saponins, ginsenoside Rg(2) [6-O-(alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl)-20(S)-protopanaxatriol] and ginsenoside Rh(1) (6-O-beta-D-glucopyranosyl-20(S)-protopanaxatriol), respectively, in high yields. Moreover, a naringinase preparation from Penicillium decumbens readily gave an intestinal bacterial metabolite, ginsenoside F(1) (20-O-beta-D-glucopyranosyl-20(S)-protopanaxatriol), as the main product, with a small amount of 20(S)-protopanaxatriol from a protopanaxatriol-type saponin mixture. Also, a hesperidinase from Penicillium sp. selectively hydrolyzed ginsenoside Re into ginsenoside Rg(1). This is the first report on the enzymatic preparation of minor saponins, ginsenosides Rg(2) and Rh(1), and of an intestinal bacterial metabolite, ginsenoside F(1), with high efficiency from a protopanaxatriol-type saponin mixture.

Enzyme Activation↗

Ginsenoside Rg8, a new dammarane-type triterpenoid saponin from roots of Panax quinquefolium.

A new dammarane-type triterpenoid saponin, ginsenoside Rg(8) (1), was isolated from the roots of Panax quinquefolium, along with five known saponins, (20E)-ginsenoside F(4) (2), ginsenosides Rh(1) (3), Rg(2) (4), F(1) (5), and (20R)-ginsenoside Rh(1) (6). The structure of ginsenoside Rg(8) (1) was determined to be (3beta,6alpha,12beta,20E)-24,25-epoxy-3,12,23-trihydroxydammar-20(22)-en-6-O-alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranoside by various spectroscopic analyses. Among the known saponins, (20E)-ginsenoside F(4) (2) and ginsenoside F(1) (5) were first reported from the title plant.

Ginsenosides↗

Somatic embryogenesis and ginsenoside production of Panax ginseng in phytohormone-free medium.

Embryogenic cultures of Panax ginseng were established without using phytohormones. Somatic embryos developed from the roots of an in vitro seedling and from excised leaf and petiole segments cultured in half-macro-salt strength Murashige and Skoog medium. Excised leaf and petiole segments were obtained from in vitro germinated seedlings. Plantlets were subsequently obtained from developing somatic embryos in phytohormone-free media. Shoot formation from somatic embryos was influenced by light intensity. The rate of growth and frequency of embryogenesis were improved when cut-up embryogenic tissues were inoculated into liquid media in the dark. The ginsenoside contents of a 4 year-old field-cultivated root, seedlings from zygotic embryos, somatic embryos and embryogenic tissues were determined and compared. Somatic embryos contained 1.7 times the amount of ginsenoside Rb1 and 2.3 times the amount of ginsenoside Re compared to seedlings from zygotic embryos. Ginsenoside Rd, which was absent in the seedlings derived from zygotic embryos, was detected in somatic embryos. Higher ginsenosides Rd and Rg1 levels were found in embryogenic tissues grown on solid media than in tissues grown in liquid media. The total ginsenoside yields, including the ginsenosides Rb1 and Rg1 levels, of cut-up embryogenic tissues, were higher than those of clump tissues.

Culture Media↗

[Combination of ginsenosides with low dose estrogen showed synergetic effect on ovariectomy induced osteopenia in rats].

AIM: To determine whether low dose of estrogen in combination with ginsenosides can completely prevent bone loss in ovariectomized rats. METHODS: Four-month-old ovariectomized rats were treated either with 100 and 300 mg.kg-1 of ginsenosides or 30 and 100 micrograms.kg-1 of 17 alpha-ethynylestradiol alone, or ginsenosides 100 mg.kg-1 in combination with 17 alpha-ethynylestradiol 30 micrograms.kg-1 for 10 weeks. Double in vivo fluorochrome labeling was made. The undecalcified longitudinal proximal tibial metaphyseal sections were processed and stained with Goldner's trichrome for histomorphometric analysis of the bone. RESULTS: Body weights and serum cholesterol were increased in ovariectomized (OVX) rats. The rats lost 74% of bone volume and high bone turnover was induced after OVX compared with the sham group. Bone volume increased by 205% in the high dose estrogen treated group while it was increased by 105% in the low dose group. The two doses of estrogen were shown to inhibit osteoclasts surface (by -65% and -55%, P < 0.01) and decrease bone turnover rate (by -85% and -83%, P < 0.01). High dose of estrogen was found to inhibit growth and stimulate uterine weight gain in rats while low dose did not. High dose of ginsenosides increased bone volume by 84% (P < 0.01) and decreased bone turnover rate by -64% (P < 0.05) while lower dose of ginsenosides did not. However, low dose ginsenosides combined with low dose estrogen achieved well preventive effects: increase of 202% in bone volume, decrease of 66% in bone turnover rate and 72% in osteoclasts surface. The combined effect in preventing bone loss equals to that the high dose of estrogen alone did. CONCLUSION: Use of low dose of estrogen plus ginsenosides showed synergistic effect on prevention of osteoporosis induced by ovariectomy.

Animals↗

[Effects of three different drying methods on extraction and separation of ginsenosides from fresh ginseng].

OBJECTIVE: There exists a close relationship between drying of a fresh herb and its preservation and extraction of efficient components. In order to investigate the influences of different drying methods on extraction and separation of ginsenosides, three drying processes, such as drying in the sun, drying in oven and microwave drying, were used to dry fresh ginsengs. METHODS: The ginsenosides of the dry ginsengs were extracted by poaching and microwave heating, and were separated by foam separation. The concentrations of ginsenosides were measured. RESULTS: Microwave drying saved both time and labor, and was favorable for release of ginsenosides. The ginsenosides could be extracted from the dry ginsengs in a shorter time by microwave heating than poaching. The ginsenosides Rb1, Rb2, Rd could be observably concentrated by foam separation. CONCLUSION: Microwave drying and microwave assisted extraction are efficient and economic methods with a high recovery yield of ginsenosides.

Desiccation↗