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Characterization of two acidic polysaccharides having immunological activities from the root of Panax ginseng.

Two acidic polysaccharides, named ginsenan PA and ginsenan PB, were isolated from the root of Panax ginseng C. A. Meyer. They were homogeneous on electrophoresis and gel chromatography, and their molecular masses were estimated to be 1.6 x 10(5) and 5.5 x 10(4), respectively. They are composed of L-arabinose: D-galactose: L-rhamnose: D-galacturonic acid: D-glucuronic acid in the molar ratios of 11:22:1:6:1 (ginsenan PA) and 3:7:2:8:1 (ginsenan PB), in addition to small amounts of O-acetyl groups. Almost all (ginsenan PA) and part (ginsenan PB) of the hexuronic acid residues exist as methyl esters. Reduction of carboxyl groups, methylation analysis, nuclear magnetic resonance and periodate oxidation studies indicated that their structural features include mainly both alpha-arabino-beta-3,6-galactan type and rhamnogalacturonan type structural units. Both polysaccharides showed remarkable reticuloendothelial system-potentiating activity in a carbon clearance test, pronounced anti-complementary activity and alkaline phosphatase-inducing activity in a dose dependent manner.

Animals↗

The core structure of ginsenan PA, a phagocytosis-activating polysaccharide from the root of Panax ginseng.

Controlled Smith degradation and limited hydrolysis of ginsenan PA, the main phagocytosis-activating polysaccharide isolated from the root of Panax ginseng C. A. Meyer, were performed. The reticuloendothelial system-potentiating and anti-complementary activities of the degradation products were investigated. Methylation analysis of the primary and secondary Smith degradation products indicated that the core structural features of ginsenan PA include a backbone chain mainly composed of beta-1,3-linked D-galactose. Almost half of the galactose units in the backbone carry side-chains composed of beta-1,6-linked D-galactosyl residues at position 6. Further 3,6-branching of D-galactose units was observed in a part of the side-chains. alpha-L-Arabinose units are connected mainly to the core galactose moieties via position 6. Removal of most of the arabinose units had a considerable effect on immunological activity.

Animals↗

Inhibition of tumor angiogenesis and metastasis by a saponin of Panax ginseng, ginsenoside-Rb2.

We studied the effect of ginsenoside-Rb2 extracted from Panax ginseng on angiogenesis and metastasis produced by B16-BL6 melanoma cells in syngeneic mice. Intravenous administration of ginsenoside-Rb2 on day 1, 3 or 7 after tumor inoculation achieved a remarkable reduction in the number of vessels oriented toward the tumor mass, but did not cause a significant inhibition of tumor growth. The anti-angiogenic effect was dose-dependent ranging from 10 to 500 micrograms/mouse. In contrast, intra-tumoral or oral administration of ginsenoside-Rb2 caused a marked inhibition of both neovascularization and tumor growth. Ginsenoside-Rb2 did not affect the growth of rat lung endothelial (RLE) cells, B16-BL6 melanoma cells or various types of murine normal cells in vitro. The invasion of RLE cells into the reconstituted basement membrane (Matrigel), which is considered to be an essential event in tumor neovascularization, was inhibited by ginsenoside-Rb2 in a concentration-dependent fashion, while ginsenoside-Rb2 did not inhibit the haptotactic migration of endothelial cells to fibronectin-substrate. Multiple administrations of ginsenoside-Rb2 after the intravenous inoculation of B16-BL6 melanoma cells resulted in a significant inhibition of lung metastasis as compared with the untreated control. These results suggest that the inhibition of tumor-associated angiogenesis by ginsenoside-Rb2 may partly contribute to the inhibition of lung tumor metastasis.

Animals↗

Inhibitory effect of ginseng total saponins on glutamate-induced swelling of cultured astrocytes.

The effects of ginseng total saponins (GTS) on L-glutamate-induced swelling of cultured astrocytes from rat brain were studied. Following exposure to 0.5 mM glutamate for 1 h, the intracellular water space (as measured by [3H]O-methyl-D-glucose uptake) of astrocytes increased three-fold with a morphological change: the disappearance of cellular processes. Simultaneous addition of GTS with glutamate reduced the astrocytic swelling in a dose-dependent manner. GTS at 0.5 mg/ml did not affect the viability of astrocytes for up to 18 h, which was determined by a colorimetric assay for cellular growth and survival. These data suggest that GTS prevents the cell swelling of astrocytes induced by glutamate.

Animals↗

Effect of ginseng radix on GLUT2 protein content in mouse liver in normal and epinephrine-induced hyperglycemic mice.

The oral administration of the water extract of Ginseng Radix (GR) to normal and epinephrine-induced hyperglycemic mice caused a significant decrease in the blood glucose level 4 h after its administration. The hepatic content of facilitative glucose transporter isoform 2, liver type glucose transporter (GLUT2) protein content from mouse liver significantly increased in the orally GR-treated normal and epinephrine-induced hyperglycemic mice compared to that in the controls. These results suggest that the hypoglycemic activity of GR is presumably due, at least in part, to the increment of GLUT2 protein content.

Animals↗

Effects of dietary supplementation of lipophilic fraction from Panax ginseng on cGMP and cAMP in rat platelets and on blood coagulation.

We have studied the effect of dietary supplementation with 25 mg (0.0025% of the total diet) of a lipophilic fraction (LF) from Panax ginseng on rat platelet aggregation induced by collagen or thrombin, and on blood coagulation. When platelets prepared from 15% corn oil plus LF-administered rats (COLF) were stimulated by thrombin (0.1 units/ml) and collagen (100 micrograms/ml), the cGMP level was significantly increased as compared with those from 15% corn oil only-administered rats (CO). The levels of cAMP in COLF were decreased by thrombin, but was increased by collagen. Furthermore, the levels of both cGMP and cAMP were also increased by the exogenous addition of LF to thrombin- and collagen-stimulated platelets. These results mean that LF increases cGMP directly and cAMP indirectly, and thus inhibits thrombin- or collagen-induced rat platelet aggregation. Both the thrombin time (TT) and activated partial thromboplastin time (APTT) were prolonged more in citrated platelet-poor plasma from COLF than in that from CO. The level of lipids such as triglyceride, total cholesterol, high density lipoprotein-cholesterol and low density lipoprotein-cholesterol was decreased in serum from COLF more than in that of CO. Thus, these results suggest that dietary LF regulates the levels of cGMP and cAMP, and prolongs the time interval (TT, APTT) between the conversion of fibrinogen to fibrin. Accordingly, our data demonstrate that dietary LF has an antithrombotic effect in vivo.

Animals↗

Specificities of five kinds of antisera produced against crude drugs, pinella tuber, hoelen, glycyrrhizae radix, trichosanthes root and panax ginseng.

Five kinds of rabbit antisera produced against five kinds of crude drugs, Pinella Tuber, Glycyrrhizae Radix (GR), Trichosanthes Root, Hoelen, and Panax Ginseng, and five kinds of extracts of the same crude drugs were prepared. Specificity of each antiserum was demonstrated by two immunological methods for analyses, the selected antibody enzyme immunoassay (SAEIA) and western blotting, and five crude drug extracts as specimens. Each crude drug extract contained characteristic antigen specific to the corresponding antiserum. Characteristic antigens were suggested as protein components. Characteristic antigen of the crude drug GR was separated from GR extract by three chromatographic procedures and a protein component was separated. The separated protein was successfully applied to develop a SAEIA method applicable for specific assays of both the separated protein and GR extract.

Animals↗

An herbal prescription, S-113m, consisting of biota, ginseng and schizandra, improves learning performance in senescence accelerated mouse.

The effect of an herbal prescription, S-113m, consisting of biota, ginseng and schizandra, on learning and memory performance was studied in the senescence accelerated mouse (SAM). A solid diet containing 1% (w/w) S-113m was given to SAM from 1 month of age. A behavioral experiment, started 4 or 9 months later, revealed prominent learning impairment in SAMP8, a senescence accelerated-prone mouse. Chronic ingestion of S-113m improved the memory retention disorder of SAMP8 in a passive avoidance test and increased the conditioned avoidance rate in a lever-press test at the age of 10 months. The preparation also facilitated the memory retention deficit in the passive avoidance test in 10-month-old SAMR1, a senescent resistant substrain. These results raise the possibility that S-113m might be useful for treating physiological aging and age-related memory deficits in human.

Aging↗

In vitro conversion of 2,3-oxidosqualene into dammarenediol by Panax ginseng microsomes.

(RS)-[3-3H]-2,3-Oxidosqualene (1) was converted into (20S)-dammarenediol (2) and not to (20R)-dammarenediol by a microsomal fraction prepared from the hairy root of Panax ginseng. The enzyme activity was highest at pH 6.0 and was not increased by the addition of any detergents. These properties differed significantly from those of other 2,3-oxidosqualene cyclases reported from higher plants and animals.

Cyclization↗

Synthesis of a biologically active fluorescent derivative of GM1, a main Ginseng saponin metabolite formed by intestinal bacteria.

A fluorescent derivative of GM1 [20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol], a main Ginseng saponin metabolite formed by intestinal bacteria, was obtained from the condensation of its trisnor-aldehyde derivative with dansyl hydrazine. The dansylated GM1 fluoresced strongly and showed almost the same properties as its parent compound in lipophilicity and biological activities, so this fluorescent compound might provide an insight into the mechanism of pharmacological activities of GM1.

Antineoplastic Agents↗

Effect of Radix Ginseng and Radix Trichosanthis on the melanogenesis.

Melanogenesis is a well known physiological response of human skin exposed to ultraviolet light, genetic reasons and other sources. In this study, we conducted to evaluate the effects of Radix Ginseng (RG) and Radix Trichosanthis (RT) on the melanogenesis in the B16 melanoma cells. The cells were treated for 48 h with RT at concentrations ranging from 1 to 50 microg/ml, RG at concentration of 10-1000 microg/ml, or RG at various doses (10-1000 microg/ml) with 25 microg/ml RT. Treatment with RT alone dose-dependently suppressed tyrosinase activity and melanin content compared with untreated control, and significantly inhibited cell proliferation. However, RG at various concentrations did not exhibit any significant change of them. Treatment with RT in the presence of various concentrations of RG suppressed tyrosinase activity and melanin content, similar to treatment with RT alone, but slightly increased cell proliferation. Furthermore, tyrosinase protein level was significantly decreased in treatment with 25 microg/ml RT alone and with a combination of 100 microg/ml RG. These results indicate that treatment with RG and RT significantly inhibits the melanogenesis in B16 cells, and raise the possibility that this combination may be effective in the whitening agent for the skin.

Animals↗

Social isolation stress-induced oxidative damage in mouse brain and its modulation by majonoside-R2, a Vietnamese ginseng saponin.

Stressors with a physical factor such as immobilization, electric foot shock, cold swim, etc., have been shown to produce oxidative damage to membrane lipids in the brain. In this study, we investigated the effect of protracted social isolation stress on lipid peroxidation activity in the mouse brain and elucidated the protective effect of majonoside-R2, a major saponin component of Vietnamese ginseng, in mice exposed to social isolation stress. Thiobarbituric acid reactive substance levels, one of the end products of lipid peroxidation reaction, were increased in the brains of mice subjected to 6-8 weeks of social isolation stress. Measurements of nitric oxide (NO) metabolites (NO(x)(-)) also revealed a significant increase of NO production in the brains of socially isolated mice. Moreover, the depletion of brain glutathione content, an endogenous antioxidant, in socially isolated animals occurred in association with the rise in lipid peroxidation. The intraperitoneal administration of majonoside-R2 (10-50 mg/kg) had no effect on thiobarbituric acid reactive substances (TBARS), NO, or glutathione levels in the brains of group-housed control mice but it significantly suppressed the increase in TBARS and NO levels and the decrease in glutathione levels caused by social isolation stress. These results suggest that mice subjected to 6-8 weeks of social isolation stress produces oxidative damage in the brain partly via enhancement of NO production, and that majonoside-R2 exerts a protective effect by modulating NO and glutathione systems in the brain.

Animals↗

Inhibition of benzo(a)pyrene induced lung adenoma by panax ginseng extract, EFLA400, in Swiss albino mice.

In the present investigation the chemopreventive action of Panax ginseng extract, EFLA400, in Swiss albino mice has been evaluated. We used a 9-week medium term anticarcinogenicity test model of lung adenomas [Yun et al.1)]. Lung adenomas were induced by single subcutaneous injection in the subscapular region with 0.02 ml of benzo(a)pyrene (BP) (0.5 mg suspension in 1% aqueous gelatin) in newborn mice (less than 24 h old). Also chromosomal aberrations and micronuclei induction were evaluated in bone marrow cells. These genotoxicity end-points were compared with adenoma incidence at the same dose levels of BP and EFLA400. The oral administration of EFLA400 (10 mg/kg body weight) showed significant reduction in number of adenomas and weight of the lungs induced by BP. A significant reduction (p<0.001) in lung adenoma incidence in EFLA400-treated mice was observed as compared to the 68.3+/-2.96% lung adenoma incidence in BP-alone group. The inhibition rate was 72.05+/-1.36% in EFLA400-treated group with respect to the reference group (BP-alone group). However, tumor multiplicity was observed as 0.91+/-0.08 and 0.25+/-0.01 in BP alone and BP+EFLA400-treated groups respectively. In EFLA400-treated group significantly reduced frequencies of chromosomal aberrations and micronuclei induced by BP were observed. The results of the present investigation suggest the chemopreventive action and antimutagenic effect of EFLA400 in Swiss albino mice induced by BP in newborn mice.

Adenoma↗

Effects of a hypoglycemic component of ginseng radix on insulin biosynthesis in normal and diabetic animals.

DPG-3-2, a component of ginseng radix, which lowers the blood glucose level and stimulates insulin release in diabetic animals, was studied for its effects on insulin biosynthesis in different preparations of pancreas from animals with normoglycemia and with hyperglycemia (alloxan diabetic rats and genetically diabetic mice, KK-CAy). We measured incorporation of radioactive leucine into insulin and other protein fractions during a 2-h perfusion of rat pancreas, and into insulin during a 3-h incubation of mouse islets. Biosynthesis of insulin during a long-term culture of islets from KK-CAy mice was also measured. DPG-3-2 was found practically not to increase the incorporation into insulin in the pancreatic preparations from animals with normoglycemia, but in such preparations from animals with hyperglycemia DPG-3-2 (0.2-1.0 mg/ml) caused 1.5-1.8-fold incorporation into insulin. In addition, a long-term treatment of DPG-3-2 (0.5 mg/ml) was shown to stimulate insulin biosynthesis in islets from KK-CAy mice. Ginsenoside-Rb1 and -Rg1 decreased the insulin content of islet to the undetectable level. Thus, DPG-3-2 was shown to stimulate insulin biosynthesis in different preparations of pancreas from animals with hyperglycemia.

Animals↗

Effect of ginseng saponins on the survival of cerebral cortex neurons in cell cultures.

The effects of nerve growth factor (NGF) and saponins isolated from Panax ginseng C.A. Mayer on the survival of chick and rat embryonic cerebral cortex neurons were examined. Ginsenoside Rg1 (GRg1) exerted a survival-promoting effect on both chick and rat cerebral cortex neurons in cell cultures. Ginsenoside Rb1 (GRb1) also had an effect in the rat and displayed some influence in the chick. NGF alone exerted no effect on both neurons, although it did potentiate the GRb1 effect on chick embryonic cerebral cortex neurons, but did not alter the GRb1 effect on rat embryonic cerebral cortex neurons. NGF did not alter the survival-promoting effect of GRg1 on either chick or rat embryonic cerebral cortex neurons. The other saponins alone or with NGF exerted no effect on the survival of cerebral cortex neurons in either the chick or rat.

Animals↗

Studies on the sesquiterpenoids of Panax ginseng C. A. Meyer. IV.

A tricarbocyclic sesquiterpenoid (1), isolated from the ethereal extract of the rootlets of Panax ginseng C.A. Meyer, showed almost identical nuclear magnetic resonance data to those reported for senecrassidiol (2). Detailed spectral analysis of 1 led us to revise the stereochemistry of 2.

Chemical Phenomena↗

Synthesis and absolute configurations of the cytotoxic polyacetylenes isolated from the callus of Panax ginseng.

Panaxacol (1) and dihydropanaxacol (2), cytotoxic polyacetylenes isolated from the callus of Panax ginseng, were synthesized starting from D-(-)-diethyl tartrate. The absolute configuration of 1 was determined to be 9R, 10R and the absolute configuration at C-3 of 2 was tentatively assigned as 3S by the application of the R(+)-alpha-methoxy-alpha-(trifluoro methyl)phenylacetyl (MTPA) method.

Alkynes↗

Cytotoxic activity of polyacetylene compounds in Panax ginseng C. A. Meyer.

The effects of the three polyacetylene compounds, panaxynol, panaxydol and panaxytriol, on in vitro-cell growth were studied. These compounds are much different in their water-solubility. In order to increase water-solubility, solid complexes of polyacetylene compounds with alpha-cyclodextrin (CD) were prepared. Accurate concentrations of the active compounds in a culture medium were determined by gas chromatography. All of these CD-complexes inhibited the growth of various kinds of cultured cell lines in a dose-dependent fashion. The cell growth inhibitory activity of these complexes was much stronger against malignant cells than against normal cells. A continuous contact between the compounds and target cells was not necessarily required for growth inhibition. And the inhibition was cytotoxic at high concentrations and cytostatic at low concentrations. These findings indicate that these polyacetylene compounds' mode of action is more dose-dependent than time-dependent. The panaxynol, panaxydol and panaxytriol contents in red ginseng powder were 250, 297 and 320 micrograms/g, respectively.

Alkynes↗