ABSTRACT
P-1 was partially hydrolyzed with 0.01, 0.05, and 0.1 M trifluoroacetic acid (TFA), successively, and the dialyzable (E-1, E-2, and E-3) and non-dialyzable (I-1, I-2, and I-3) fractions were prepared and analyzed chemically and immunochemically. Either I-1 or E-1 reacted with anti P-1 serum as strongly as P-1 and were mitogenic. The cross-reactivity of I-2 and I-3 was less than I-1 with anti P-1 serum. However, they were as mitogenic as I-1. The cross-reactivity of E-2 and E-3 to anti P-1 serum was also very weak, and they were not mitogenic. The E-1 fraction had a similar sugar composition to I-1 and P-1. E-2 was a monosaccharide, all of Ara, and would be from the linkage of furanosyl residues in P-1. The composition of E-3 was free from Ara and the structure of E-3 was similar to that of I-3. E-3 would be considered to be deleted arabinofuranose from E-1. These results suggest that the mitogenic activity measured by the alkaline phosphatase assay is a property of the core part, I-3, but that P-1 contains several epitopes other than the core part by the immunochemical analysis.
Subject(s)
Fruit/chemistry , Plants, Medicinal/chemistry , Polysaccharides/chemistry , Animals , Antibody Specificity , Carbohydrate Sequence , Chemical Phenomena , Chemistry, Physical , Hydrogen-Ion Concentration , Hydrolysis , Immunochemistry , Japan , Mice , Mice, Inbred C3H , Mitogens/pharmacology , Molecular Sequence Data , Oxidation-Reduction , Polysaccharides/pharmacology , Rabbits/immunology , Spleen/cytology , Spleen/drug effectsABSTRACT
The fruits of Prunus mume Sieb. et Zacc. (Japanese name, ume) have been used as a traditional drug and health food. In order to study the active components of P. mume, the polysaccharide fractions were extracted with cold water, hot water and aqueous sodium hydroxide from the kernels of P. mume. We found that some of the polysaccharide fractions exhibited various types of biological activities such as mitogenesis, activation of the alternative pathway of complement and activation of clot formation in human plasma. A polysaccharide, P-1, obtained from the cold 0.5 M NaOH extract was purified by ion-exchange chromatography and gel-filtration, P-1 contained 62.0% neutral sugar as glucose and 38.4% uronic acid (as galacturonic acid), and was free from protein. The neutral sugars of P-1 were arabinose, xylose, rhamnose and galactose in a molar ratio of 9.4:3.4:1.1:1.0, following analysis by gas-liquid chromatography. In addition, galacturonic acid was identified by thin-layer chromatography. The molecular weight of P-1 was found to be more than 2,000,000 by gel-filtration on Toyopearl HW 65F. P-1 showed mitogenic activity towards spleen cells of both C3H/HeN and C3H/HeJ, suggesting that it was free from bacterial endotoxic lipopolysaccharides.
Subject(s)
Plants, Medicinal/chemistry , Polysaccharides/pharmacology , Animals , Carbohydrates/analysis , Cells, Cultured , Chemical Phenomena , Chemistry, Physical , Chromatography, Gas , Chromatography, Ion Exchange , Chromatography, Thin Layer , Complement Pathway, Alternative/drug effects , Humans , Male , Mice , Mice, Inbred C3H , Mitogens/pharmacology , Molecular Weight , Plant Extracts/analysis , Plant Extracts/pharmacology , Polysaccharides/chemistry , Polysaccharides/isolation & purification , Spleen/cytology , Spleen/drug effects , UltracentrifugationABSTRACT
Antimutagenic activities of hexane extracts obtained from the fruit extract (UE) and the Kernels (KE) of P. mume were examined. These extracts showed inhibitory activities to known mutagens, 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide, benzo[alpha]pyrene and aflatoxin B1 in the Ames assay using Salmonella typhimurium TA100 and TA98 strains. The UE and KE fractions were then separated by silicic acid column chromatography with a stepwise elution method using ether-hexane. The location of the active substances in the fractions was also determined by thin-layer chromatography. Consequently, it was found that the effective substances for the desmutagenicity were fatty acids, and identified by gas liquid chromatography, mainly as oleic acid, linoleic acid and linolenic acid in UE, and mainly as oleic acid and linoleic acid in KE, respectively.