Inhibitory effect of tumor metastasis in mice by saponins, ginsenoside-Rb2, 20(R)- and 20(S)-ginsenoside-Rg3, of red ginseng.

We examined the inhibitory effect of two saponin preparations from Red ginseng, 20(R)- and 20(S)-ginsenoside-Rg3, in comparison with that of ginsenoside-Rb2, on lung metastasis produced by two highly metastatic tumor cells, B16-BL6 melanoma and colon 26-M3.1 carcinoma, in syngeneic mice. In an in vitro analysis, both saponin preparations showed a significant inhibition of adhesion to fibronectin (FN) and laminin (LM) by B16-BL6 melanoma. Similarly, they significantly inhibited the invasion of B16-BL6 cells into the reconstituted basement membrane (Matrigel)/FN in a dose-dependent manner. In an experimental metastasis model using B16-BL6 melanoma, consecutive intravenous (i.v.) administrations of 100 micrograms/mouse of 20(R)- or 20(S)-ginsenoside-Rg3 1, 2, 3 and 4 d after tumor inoculation led to a significant decrease in lung metastasis. The inhibitory effect of i.v. administration of both ginseng saponins on the tumor metastasis of B16-BL6 melanoma was also recognized in a low dose of 10 micrograms/mouse. The oral administration (p.o.) of both saponins (100-1000 micrograms/mouse) induced a significant decrease in lung metastasis of B16-BL6 melanoma. Moreover, both ginseng saponins were effective in inhibiting of lung metastasis produced by colon 26-M3.1 carcinoma. When 20(R)- or 20(S)-ginsenoside-Rg3 was orally administered consecutively after tumor inoculation in a spontaneous metastasis model using B16-BL6 melanoma, both of them significantly inhibited lung metastasis. In the experiment involving neovasculization by tumor cells in vivo, both mice groups given each saponin preparation after tumor inoculation exhibited a significant decrease in the number of blood vessels oriented toward the tumor mass, with no repression of tumor size. These findings suggest that both ginseng saponins, 20(R)- and 20(S)-ginsenoside-Rg3, possess an ability to inhibit the lung metastasis of tumor cells, and the mechanism of their antimetastatic effect is related to inhibition of the adhesion and invasion of tumor cells, and also to anti-angiogenesis activity.

[Simultaneous analysis of ginsenosides of various ginseng radix by HPLC].

A simultaneous analysis of ginsenosides in Panax ginseng by high performance liquid chromatography recently established by us was applied to the analysis of various Ginseng Radix. The contents of ginsenosides in P. ginseng were examined according to the differences of the growth year, the used part of the plant, the method of processing and the cultivated location. In the case of P. ginseng cultivated in Nagano, Japan, the ratio (total ginsenosides content/total dry root weight) increases annually for three years. And it decreased at the fourth year and increases again at the fifth and the sixth years. Concerning the distribution of ginsenosides in the parts of the plant, they were contained at the highest level in a lateral root and in succession, in a rhizome > in a root hair > in a main root. They were also distributed much richer at periderm than at phloem or at xylem of a main root. The contents of panaxadiol- and panaxatriol-saponins gradually increase with the growth year, whereas an oleananesaponin, ginsenoside-Ro, drastically increases at the sixth year and goes to 15-fold, which suggests that the content of ginsenoside-Ro needs to be estimated much more when Ginseng Radix is evaluated. The fact that the processing for preparation of Red Ginseng increases the total content of saponins was clearly revealed by the present study. The highest contents of saponins among Red Ginsengs (all 6-year-old) were observed in ones prepared in Korea, and in Japan > in China, successively.

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.

Anti-hepatitic activity of ginesenoside ro1.

Ginsenoside Ro, an oleanane-type saponin has been screened for activity in experimental models of acute and chronic hepatitis. Ginsenoside Ro (50 and 200 mg/kg, P.O.) inhibited the increase of serum glutamic oxaloacetic transaminase (s-GOT) and serum glutamic pyruvic transaminase (s-GPT) levels in D-galactosamine (GalN)- and carbon tetrachloride (CCl (4))-induced acute hepatitic rats. Ginsenoside Ro inhibited the increase of connective tissue in the liver of CCl (4)-induced chronic hepatitic rats. Ginsenoside Ro showed a stronger inhibitory effect on the GalN-induced acute hepatitic model than those of the aglycone of ginsenoside Ro, oleanolic acid, or glycyrrhizic acid and its aglycone, glycyrrhetinic acid.

Anti-hepatitic activity of ginsenoside Ro.

Ginsenoside Ro, an oleanane-type saponin has been screened for activity in experimental models of acute and chronic hepatitis. Ginsenoside Ro (50 and 200 mg/kg, p.o.) inhibited the increase of serum glutamic oxaloacetic transaminase (s-GOT) and serum glutamic pyruvic transaminase (s-GPT) levels in D-galactosamine (GalN)- and carbon tetrachloride (CCl4)-induced acute hepatitic rats. Ginsenoside Ro inhibited the increase of connective tissue in the liver of CCl4-induced chronic hepatitic rats. Ginsenoside Ro showed a stronger inhibitory effect on the GalN-induced acute hepatitic model than those of the aglycone of ginsenoside Ro, oleanolic acid, or glycyrrhizic acid and its aglycone, glycyrrhetinic acid.

[Measurement of abundance ratio of 15N in amino acids by capillary gas chromatography-mass spectrometry].

A capillary gas chromatography-mass spectrometry for analysis of 15N-labeled amino acids and amides is described. The method is based on direct silylation of amino acids and amides with MTBSTFA and the formation of the TBDMS derivatives. The method was possible simultaneously to measure the 15N abundance ratio of amino-N and amide-N of amides, as to analysis of amino acids.

Anti-inflammatory activity of ginsenoside Ro.

Ginsenoside Ro, an oleanane-type saponin has been screened for activity in experimental models of inflammation. Ginsenoside Ro (10, 50, and 200 mg/kg, p.o.) inhibited an increase in vascular permeability in mice induced by acetic acid and reduced an acute paw edema in rats induced by compound 48/80 or carrageenin. Ginsenoside Ro did not suppress a developing adjuvant-induced edema in arthritic rats. However, ginsenoside Ro was found to be effective in hypercoagulable state, increase of connective tissue in the artery and calcium effluence from the bone in adjuvant-induced arthritic rats.

Anti-inflammatory activity of ginsenoside ro.

Ginsenoside Ro, an oleanane-type saponin has been screened for activity in experimental models of inflammation. Ginsenoside Ro (10,50, and 200 mg/kg, P. O.) inhibited an increase in vascular permeability in mice induced by acetic acid and reduced an acute paw edema in rats induced by compound 48/80 or carrageenin. Ginsenoside Ro did not suppress a developing adjuvant-induced edema in arthritic rats. However, ginsenoside Ro was found to be effective in hypercoagulable state, increase of connective tissue in the artery and calcium effluence from the bone in adjuvant-induced arthritic rats.

Studies of Panax japonicus fibrinolysis.

The effects of a 70% methanol extract (PMe) obtained from the rhizomes of Panax japonicus C. A. Meyer on experimental thrombosis and fibrinolysis were investigated in vivo and in vitro. PMe showed a promotive effect on the activation of the fibrinolytic system as determined by the euglobulin lysis time (ELT) assay but was inactive to the inhibitory effect against endotoxin-induced disseminated intravascular coagulation (DIC) in rats. PMe and its major components, chikusetsusaponin III, IV, and V, strongly promoted the action of urokinase in fibrin plate. These results suggested that PMe promotes the fibrinolysis and its effective components are chikusetsusaponin III, IV, and V.

Study of ammonia metabolism in a patient with ornithine transcarbamylase deficiency using an 15N tracer.

Ammonia metabolism was studied in an 8-year-old girl with ornithine transcarbamylase (OTC) deficiency, using 15N-tracer. Changes in the incorporation of 15N into amino acids and urea were examined after 15NH4Cl administration. The recovery of total 15N in the urine of the patient in 3 days was 28.5% of the administered 15N whereas that of a control was 69.3%. They were mostly urea. The recovery of 15N-urea in the patient was 28.8% of the control in 1 day, 32% in 2 days and 33.3% in 3 days after the administration of 15NH4Cl. A larger amount of 15N was incorporated into glutamine (alpha-amino N) and glutamate and 15N was incorporated more rapidly into alanine, asparagine and serine in the patient than in the control. The incorporation into ornithine was less in the patient than in the control.