sapogenins and Hemolysis

Topics: Animals; Antineoplastic Agents; Cell Death; Cell Line, Tumor; Female; Hemolysis; Hydrogen-Ion Concentration; Kinetics; Mice, Inbred C57BL; Nanoparticles; Panax; Plant Extracts; Sapogenins; Tissue Distribution

We evaluated the hemolytic activity of 41 commercially available triterpenoid saponins and sapogenins derived from three types of structural skeletons. Structure-activity relationships were established by comparing the structural characteristics of both the aglycone and sugar moieties among the tested compounds. The majority of oleanane-type sapogenins had stronger hemolytic effects than those of the ursane and dammarane types. The presence of polar regions on sapogenins, such as a carboxyl (COOH) at position 28, an α-hydroxyl (α-OH) at position 16, and/or a β-hydroxyl (β-OH) at position 2, significantly enhanced hemolysis. Meanwhile, the introduction of an α-OH at position 2 or a methyl hydroxyl (CH

Topics: Hemolysis; Sapogenins; Saponins; Structure-Activity Relationship; Triterpenes

The further purification of the total saponins from the roots of Panax notoginseng by using ordinary and reversed-phase silica-gel, as well as Sephadex LH-20 chromatography afford seven adjuvant active protopanaxatriol-type saponins (PTS), ginsenosides-Rh1 (Rh1),-Rh4 (Rh4),-Rg1 (Rg1),-Re (Re), notoginsenosides-R1 (R1),-R2 (R2),-U (U). These saponins were evaluated for their haemolytic activities and adjuvant potentials on the cellular and humoral immune responses of ICR mice against ovalbumin (OVA). The effect of the substitution pattern of these PTS on their biological activities was investigated and structure-activity relationships were established. Among seven PTS, the haemolytic activity of Rh1 was higher than that of other six compounds (p<0.001) The HD50 values of Rh4 and U were significantly bigger than those of R2, Rg1 and Re (p<0.05 or p<0.01). Seven PTS could significantly increase the concanavalin A (Con A)-, lipopolysaccharide (LPS)- and OVA-induced splenocyte proliferation in the OVA-immunized mice (p<0.01 or p<0.001). The OVA-specific IgG, IgG1, IgG2a and IgG2b antibody levels in serum were also significantly enhanced by seven PTS compared with OVA control group (p<0.01 or p<0.001). The structure-activity relationship studies suggested that the number, the length and the position of sugar side chains, and the type of glucosyl group in the structure of PTS could not only affect their haemolytic activities and adjuvant potentials, but have significant effects on the nature of the immune responses. The information about this structure/function relationship might be useful for developing semisynthetic tetracyclic triterpenoid saponin derivatives with immunological adjuvant activity, as well as a reference to the distribution of the functional groups composing the saponin molecule.

Topics: Animals; Antibody Formation; Dose-Response Relationship, Drug; Hemolysis; Immunity, Cellular; Immunologic Factors; In Vitro Techniques; Lymphocyte Activation; Male; Mice; Mice, Inbred ICR; Ovalbumin; Panax; Plant Roots; Sapogenins; Saponins; Structure-Activity Relationship; Triterpenes

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 gi

Topics: alpha-Tocopherol; Amidines; Antioxidants; Drug Synergism; Erythrocytes; Ginsenosides; Hemolysis; Humans; Inhibitory Concentration 50; Membrane Lipids; Molecular Structure; Sapogenins; Saponins; Time Factors; Triterpenes

Sapogenins from the starfish Asterias amurensis and Lethasterias nanimensis chelifera, 5 alpha-pregn-9(11)-ene-3 beta,6 alpha-diol-20-one, 5 alpha-cholest-9(11)-ene-3 beta,6 alpha-diol-23-one, 5 alpha-cholesta-9(11),24(25)-diene-3 beta,6 alpha-diol-23-one, (20E)-5 alpha-cholesta-9(11),20(22)-diene-3 beta,6 alpha-diol-23-one and 24 zeta-methyl-5 alpha-cholesta-9(11),20(22)-diene-3 beta,6 alpha-diol-23-one, stimulated the contractile force of the heart of the mollusk Spisula sachalinensis at concentration of 5 x 10(-5) M. Ouabain, a specific inhibitor of Na+,K(+)-ATPase, at concentration of 5 x 10(-5) M had no effect on this physiological model. Starfish sapogenins of the cholestane series moderately inhibited rat brain cortex Na+,K(+)-ATPase and decreased Ca2+ influx into Ehrlich carcinoma cells. In contrast, pregnane asterogenin asterone did not inhibit Na+,K(+)-ATPase and increased the influx of Ca2+ into cells. These effects were not the result of cell membrane damage, because none of the compounds tested have hemolytic activity.

Topics: Animals; Brain; Calcium; Cell Membrane; Cholestenones; Enzyme Inhibitors; Heart; Hemolysis; Humans; Mollusca; Myocardial Contraction; Ouabain; Pregnenes; Rats; Sapogenins; Saponins; Sodium-Potassium-Exchanging ATPase; Starfish; Sterols; Tumor Cells, Cultured

Kleingrass (Panicum coloratum L) has been reported to cause hepatogenous photosensitization in sheep and goats in West Texas since 1973. The toxin in Kleingrass has been suspected of being a steroidal saponin as Kleingrass produced characteristic birefringent crystals similar to those produced by saponin-containing plants such as Agave lecheguilla and Tribulus terrestris. In this present study, steroidal saponis were isolated from Kleingrass and their sapogenins were identified as diosgenin and yamogenin by means of thin-layer chromatography and infrared spectrophotometric analysis.

Topics: Chromatography, Thin Layer; Diosgenin; Edible Grain; Hemolysis; Panicum; Sapogenins; Saponins; Spectrophotometry, Infrared

The hemolytic properties and the adsorbability on red blood cells of saikosaponin a, saikosaponin d and 13 metabolites formed in the alimentary tract were investigated. Among these compounds, saikosaponin d and its intestinal product, prosaikogenin G, which possess an alpha-hydroxyl function at C16, showed the strongest hemolytic activity at the dose range of 1.0 to 5.0 micrograms/ml. Saikosaponin a and its intestinal product, prosaikogenin F, which possess a beta-hydroxyl function at C16, showed activity above 10 micrograms/ml. In this case, the monoglycoside, prosaikogenin F, showed the stronger activity than the diglycoside, saikosaponin a. Among the gastric products whose ether ring was cleaved to produce a carbinol, the monoglycosides, prosaikogenin A and prosaikogenin H, showed a slight activity above 25 micrograms/ml, and the saikogenins except saikogenin A were inactive. Saikogenin A, however, had hemolytic activity at a dose of 15 micrograms/ml. The adsorbabilities of these compounds on red blood cell membranes closely paralleled their degrees of hemolytic activity.

Topics: Adsorption; Erythrocyte Membrane; Hemolysis; Humans; In Vitro Techniques; Oleanolic Acid; Sapogenins; Saponins

Topics: Anti-Inflammatory Agents, Non-Steroidal; Hemolysis; Hot Temperature; Medicine, Chinese Traditional; Medicine, East Asian Traditional; Oleanolic Acid; Plants, Medicinal; Sapogenins; Saponins

Topics: Animals; Chemical Phenomena; Chemistry; Ginsenosides; Hemolysis; In Vitro Techniques; Oleanolic Acid; Panax; Plants, Medicinal; Sapogenins; Saponins; Sheep

Effects of saponins extracted from Bupleuri Radix (saikosaponin) and the corresponding aglycones on hypotonic or hyperthermic hemolysis were investigated. Low concentrations of saikosaponins protect or stabilize rat erythrocytes against both hypotonic and heat-induced hemolysis. Minor modifications of the aglyconic part of the saikosaponin have enormous effects on the membrane stabilizing potency. Saikogenins also protect erythrocytes from hypotonic hemolysis but do not show any prevention of heat-induced hemolysis. It is suggested that saikogenins react with erythrocyte membranes in a quite different manner from saponins and that the existence of the sugar moiety plays an important role in the reaction with membranes as does a slight modification of the molecular structure in the aglyconic part.

Topics: Animals; Erythrocyte Membrane; Erythrocytes; Hemolysis; Hot Temperature; In Vitro Techniques; Male; Oleanolic Acid; Osmolar Concentration; Rats; Sapogenins; Saponins; Structure-Activity Relationship

The hemolytic properties of nonionic surfactants of the series CH3(CH2)15-17-O-(CH2CH2O)nCH2CH2OH were investigated and compared to those of saponins, sapogenins and H2O2. Antioxidants and anaerobic conditions were shown to inhibit the hemolysis, while glycyrrhizin was found to enhance it. Similar effects were obtained for H2O2 hemolysis, but not for saponin and sapogenin hemolysis. It is proposed that peroxides and free radicals are mainly responsible for the polyoxyethylene derived surfactants induced hemolysis.

Topics: Animals; Cetomacrogol; Glycyrrhetinic Acid; Glycyrrhizic Acid; Hemolysin Proteins; Hemolysis; Peroxides; Polyethylene Glycols; Rats; Sapogenins; Saponins; Surface-Active Agents

Chikusetsusaponin IV and V, whose genin is oleanolic acid, exhibited weak hemolytic activities. Removal of glucose residue at position 29 of chikusetsusaponin V by partial hydrolysis increased the activity more than 30-fold. Methylation of the carboxyl group at position 28 increased the activity furthermore by about 10-fold, showing HD50 value of 3.77 microM. On the other hand, removal of the sugar chain at position 3 of chickusetsusaponin V by partial hydrolysis completely lost the activity. These facts suggest that the sugar chain at position 3 of oleanolic acid is essential but that at position 29 is pernicious for the activity. The cytolytic agents, whose target has been regarded as membrane cholesterol, were inactivated not only by cholesterol but also by sapogenins such as oleanolic acid, gitogenin and hederagenin. Among saponins tested, akebia saponin B and C were inactivated by cholesterol, but not by the genins, probably because their affinities for the genins are too low to form complexes.

Topics: Anti-Bacterial Agents; Chemical Phenomena; Chemistry; Cholesterol; Hemolysis; Humans; In Vitro Techniques; Oleanolic Acid; Panax; Plants, Medicinal; Sapogenins; Saponins; Structure-Activity Relationship

Clinical observations on four male cow calves and three other lantana poisoned animals under field conditions have been made along with hematological studies, including observations on plasma bilirubin content and osmotic fragility of erythrocytes. Four healthy male cow calves served as control. Five clinically advanced cases succumbed within 1 to 10 days of observation. Hematocrit values in four of the seven affected animals were abnormally high. The total plasma bilirubin content in the fatal cases was of the order of 5 to 50 times the normal value while, in calves that recovered, it was not more than 3 times. The osmotic fragility of erythrocytes was measured by the rate of hemolysis in decreasing concentrations of saline. The erythrocytes of normal animals resisted hemolysis till the saline concentration was lowered to 0.60%, and it was complete at the low saline concentration of 0.30%. However, in lantana poisoned animals, hemolysis started even in 0.72% saline and was complete at a saline concentration of 0.44%. The hemolysis curves of animals that died were far removed from the normal, but those of the calves that recovered and the one that died but had the lowest bilirubin content closely approached the normal curve.

Topics: Animals; Cattle; Cattle Diseases; Erythrocyte Membrane; Erythrocytes; Female; Hemolysis; Male; Oleanolic Acid; Osmotic Fragility; Plant Poisoning; Plants, Toxic; Sapogenins

The influence of vitamin E (9, 90, 450 mg dl-alpha-tocopherol acetate/100 g diet), protein quality and temperature (70 degrees C for 10 min) on the sapogenin-induced hemolysis was studied in rats fed with diets containing different amounts of fenugreek-seed seedflour. Significant increased hemolysis rates were found in rats on diets with the following protein components: 1/3 Trigonella + 2/3 Cornflour (69.7 +/- 11.6% hemolysis), 100% Trigonella (21.0 +/- 8.6% hemolysis), 1/3 Trigonella + 2/3 Casein (14.1 +/- 10.7% hemolysis). The diets of these groups of animals contained raw Trigonella seeds and 9 mg vitamin E/100 g diet, which covers the requirements of rats on this vitamin. With this tocopherol supplement the applied heat treatment had no influence on the sapogenin effects on the erythrocytes. In the groups with the 10th fold and 50th fold of the vitamin E supplementation no significant hemolysis rate was observed. From these results we conclude that when fenugreek seeds are to be used in animal nutrition, a qualitative good protein supply as well as a sufficient intake of vitamin E should be considered in order to compensate the hemolytic effect of Trigonella sapogenins.

Topics: Animals; Dietary Proteins; Female; Hemolysis; Plants; Rats; Sapogenins; Species Specificity; Vitamin E

The assumption that complex formation between erythrocyte membrane cholesterol and saponins or sapogenins is the cause for their hemolytic activity, was tested by measuring the susceptibility of cholesterol-depleted erythrocytes towards these hemolysins. For some of the hemolysins cholesterol depletion caused inhibition of hemolysis, for others an augmentation. The results suggest that cholesterol does not serve as a specific binding site for these hemolysins.

Topics: Animals; Cholesterol; Erythrocyte Membrane; Erythrocytes; Hemolysis; Male; Membrane Lipids; Rats; Sapogenins; Saponins

Topics: Cell Death; Hemolysis; Humans; Sapogenins; Saponins

Topics: Cell Death; Hemolysis; Humans; Sapogenins; Saponins