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Minimizing the saponin content of alfalfa sprouts and leaf protein concentrates.

Biologically active saponins were found to be concentrated in the white and green alfalfa leaf protein fractions at levels higher than in the original alfalfa. Coagulation and washing of the leaf protein at pH 8.5 resulted in a fourfold decrease in saponin compared to the protein coagulated at pH 6.0 and washed at pH 4.5. The press cakes from high and low saponin alfalfa retained 65 and 87% of the saponin. Protein fractions prepared from a low saponin alfalfa contained saponin levels less than 0.07%, compared to a saponin level of 1.33% in leaf protein prepared form a high saponin alfalfa. The saponin contents of three varieties of alfalfa sprouts ranged from 1.55 to 7.27% depending upon the maturity of the sprouts. The saponin content rapidly increased after sprouting and reached a maximum after eight days' growth. Both starch and total sugar decreased with the age of the sprouts while fiber increased from 8 to 18.7%.

Animals↗

Role of transforming growth factor-beta pathway in the mechanism of wound healing by saponin from Ginseng Radix rubra.

1. The effects of saponin from Ginseng Radix rubra on extracellular matrix metabolism, the activation and synthesis of TGF-beta1, and the modification of TGF-beta receptor in fibroblasts were examined to elucidate the contribution of the TGF-beta pathway to the mechanism of wound healing by saponin. 2. Fibronectin synthesis was analysed by the immunoprecipitation method. Activation and synthesis of TGF-beta1 were measured by ELISA. The expressions of TGF-beta receptors in fibroblasts were examined at protein and mRNA levels by the cross-linking method and Northern blot analysis, respectively. 3. The fibronectin synthesis increased 2.3- and 3.9-fold at fibroblasts treated with 1 and 10 microg ml(-1) of saponin, respectively, compared with that in non-treated cells. Fibronectin synthesis stimulated with 10 microg ml(-1) of saponin was inhibited with 69% by 5 microg ml(-1) of an anti-TGF-beta1 antibody. mRNA of TGF-beta type I receptor increased 4.8- and 4.4-fold at fibroblasts treated with 1 and 10 microg ml(-1) of saponin, respectively, and that of TGF-beta type II receptor also increased 3.4- and 3.2-fold at fibroblasts treated with 1 and 10 microg ml(-1) of saponin, respectively. The significant increases of TGF-beta type I and II receptors and of fibronectin synthesis were observed at the same concentrations of saponin. TGF-beta content increased 1.74- and 1.87-fold at conditioned medium of fibroblasts treated with 100 and 250 microg ml(-1) of saponin, respectively, higher concentrations than those which accelerated fibronectin synthesis. Furthermore, the active TGF-beta content was below 10% of total TGF-beta at each concentration of saponin. 4. These results indicate that saponin stimulates fibronectin synthesis through the changes of TGF-beta receptor expressions in fibroblasts.

Antibodies↗

Stimulation of LH release from cultured pituitary cells by saponins of Petersianthus macrocarpus: a permeabilizing effect.

Aqueous extracts from stem bark of Petersianthus macrocarpus contain substances exhibiting both estrogenic and antiestrogenic potency. Triterpenic saponins were identified and extracted as a bulk. Their action on the in vitro LH released by cultured rat pituitary cells was investigated. P. macrocarpus saponins stimulated the LH release in a dose-dependent manner (from 10 micrograms/ml to 300 micrograms/ml). When added simultaneously, saponins and LHRH exerted initial additive effects on LH release, demonstrating independent mechanisms of stimulation. If cells were pre-treated with saponins for 15 min, the amount of LH released under a subsequent LHRH stimulation was lowered, presumably due to a partial depletion of the cells in hormone (data not presented). However, the action of saponins on LH release did not appear specific since a general permeabilizing effect of the cell membrane was evidenced both by trypan blue exclusion and by analysis of the total protein output. When using low concentrations of saponins (10 micrograms/ml), scanning electron microscopy did not reveal any significant alteration of the cell structure, which explains why the cells remain responsive to LHRH after withdrawal of saponins. With higher concentrations (greater than 30 micrograms/ml), the same analytical studies evidenced numerous perforations of the cell membrane, with subsequent cell death. Two highly purified saponin species were tested on LH release by cultured cells; one of them (petersaponin I) appeared responsible for the observed biological effects in vitro. But as cells were shown to be efficiently protected from saponin effects by the presence of serum, it may be concluded that saponins of P. macrocarpus extracts are probably not candidate molecules promoting the in vivo estrogenic and antiestrogenic effects.

Animals↗

Effects of alfalfa root saponins on digestive function in sheep.

The effects of alfalfa saponins on ruminal pH and VFA concentrations, protozoal numbers, forestomach motility, and digesta flow from the rumen were examined in this experiment. In a preliminary study, either 800 or 1600 mg/kg of saponins per kg BW were administered intraruminally in a single dose to one of two 60-kg wethers. Ruminal contractions were suppressed within 15 min. The study was terminated when one wether didn't recover. Saponins were then administered intraruminally in two equal doses daily into four ruminally and duodenally cannulated wethers (60 +/- 1 kg) in amounts equivalent to 0, 200, 400, and 800 mg/kg BW daily in a 4 x 4 Latin Square arrangement. Saponins increased VFA concentrations (P = .001) and lowered ruminal pH (P = .001) after 2 d of administration. On d 2 and d 14, saponins decreased (P < .01) protozoal populations. On d 11, pressure changes during ruminal contractions increased by 79% during resting (P = .06), 202% during feeding (P = .05), and 53% during an 8-h period (P = .08) as the level of saponins increased from 0 to 800 mg/kg BW. Administered saponins increased duodenal flows of OM and total N by 19 and 26%, respectively (P = .04 and .01). Apparent total tract N digestibility was reduced (P = .02) by 5% at the highest saponin dosage. We conclude that alfalfa may contain enough saponins to adversely impact ruminoreticular motility. Saponins also caused decreased ruminal protozoal populations, increased flow of total duodenal N, and reduced ruminal and total tract apparent digestibilities.

Animals↗

Metabolomic analysis of saponins in crude extracts of Quillaja saponaria by liquid chromatography/mass spectrometry for product authentication.

Analysis of 50% aqueous methanolic extracts of bark of Quillaja saponaria Molina (quillaja) by liquid chromatography/mass spectrometry (LC/MS), using negative ion electrospray, revealed over 100 saponins. The majority could be assigned to known structures or generalised variations of these from the product ion spectra obtained by serial mass spectrometry in a quadrupole ion trap mass spectrometer. Ten saponins contained a fatty acid domain terminated with both a pentose and deoxyhexose unit, a feature thus far only reported in QS-III. Twenty saponins were based on a hydroxylated derivative of quillaic acid, whereas only six 22beta-hydroxyquillaic acid saponins have been described. The occurrence of pairs of saponins differing only by the presence of a rhamnose or xylose unit in the C-3-substituted saccharide was readily observed in two-dimensional mass maps, and these showed the presence of the unreported 'rhamnose partner' of QS-III. However, one sample labelled as Q. saponaria appeared to lack all saponins containing rhamnose in the C-3 saccharide. Methods to authenticate saponin extracts of quillaja by LC/MS are suggested based on the general metabolomic profile, the occurrence of specific major saponins covering known structural variations, or the presence of saponins containing the unusual fatty acid domain, revealed by neutral loss analysis.

Chromatography, High Pressure Liquid↗

Fungitoxic activity of saponins: practical use and fundamental principles.

Saponins, as aescin or alfalfa saponins have fungistatic activity on Trichoderma viride G fungal strain. (Biacs and Gruiz, 1982) The principle of fungistatic activity is an interaction between saponin and membrane constituents, such as sterols, proteins and phospholipids. The interaction results in the destruction of the cell membrane and an increase of the ion permeability (Gruiz and Biacs, 1989). Some Trichoderma strains are sensitive, whereas others are resistant to saponins. Their lipid composition after treatment differs considerably (Biacs and Gruiz, 1984), although ergosterol content of the membrane increased in both sensitive and nonsensitive strains. The possible metabolic changes in fatty acid composition of membrane phospholipids influence sensitivity of fungi to saponins. An important role is attributed to linoleic acid (Gruiz and Biacs, 1989). Since the fungal membrane composition can be changed by exogenously added sterols and fatty acids or triglycerides (Gruiz and Biacs, 1990), experiments were made to decrease saponin sensitivity of Trichoderma viride by giving linoleic-acid-containing oil and ergosterol. In such a way the elimination of the inhibitory effect of saponins was successful. The ergosterol content after saponin treatment is almost equal value not depending on the sensitivity and the ergosterol content of control or modified cells. This equalizing behavior of saponins is remarkable; its regulatory effect affects not only the ergosterol content, but also the fatty acid composition of the fungal membrane.

Antifungal Agents↗

Effect of calmodulin on ginseng saponin-induced Ca2+-activated Cl- channel activation in Xenopus laevis oocytes.

We previously demonstrated the ability of ginseng saponins (active ingredients of Panax ginseng) to enhance Ca2+-activated Cl- current. The mechanism for this ginseng saponin-induced enhancement was proposed to be the release of Ca2+ from IP3-sensitive intracellular stores through the activation of PTX-insensitive Galpha(q/11) proteins and PLC pathway. Recent studies have shown that calmodulin (CaM) regulates IP3 receptor-mediated Ca2+ release in both Ca2+-dependent and -independent manner. In the present study, we have investigated the effects of CaM on ginseng saponin-induced Ca2+-activated Cl- current responses in Xenopus oocytes. Intraoocyte injection of CaM inhibited ginseng saponin-induced Ca2+-activated Cl- current enhancement, whereas co-injection of calmidazolium, a CaM antagonist, with CaM blocked CaM action. The inhibitory effect of CaM on ginseng saponin-induced Ca2+-activated Cl- current enhancement was dose- and time-dependent, with an IC50 of 14.9 +/- 3.5 microM. The inhibitory effect of CaM on saponin's activity was maximal after 6 h of intraoocyte injection of CaM, and after 48 h the activity of saponin recovered to control level. The half-recovery time was calculated to be 16.7 +/- 4.3 h. Intraoocyte injection of CaM inhibited Ca2+-induced Ca2+-activated Cl- current enhancement and also attenuated IP3-induced Ca2+-activated Cl- current enhancement. Ca2+/CaM kinase II inhibitor did not inhibit CaM-caused attenuation of ginseng saponin-induced Ca2+-activated Cl- current enhancement. These results suggest that CaM regulates ginseng saponin effect on Ca2+-activated Cl current enhancement via Ca2+-independent manner.

Animals↗

Quantitative determination of the group of flavonoids and saponins from the extracts of the seeds of Glinus lotoides and tablet formulation thereof by high-performance liquid chromatography.

The total flavonoids and saponins of the seeds of Glinus lotoides in the crude extracts and tablet formulation thereof were quantified by reversed-phase high-performance liquid chromatographic (RP-HPLC) methods with UV detection. The saponins were analyzed after acid hydrolysis in 3 M HCI at 100 degrees C for 1 h. Vicenin-2 and mollugogenol B were isolated and used as reference substances for the quantification of total flavonoids and saponins, respectively. The identity and purity (> 97%) of the standards were confirmed by spectroscopic (UV, MS, and NMR) and chromatographic (HPLC) methods. The flavonoids and saponins of the crude extract of the seeds and tablet formulation were separated by RP-HPLC (Nucleosil RP-18 column, 250 mm x 4.6 mm) using linear gradient elution systems of acetonitrile-water-0.1 M H3PO4 for flavonoids and methanol-water for saponins. Satisfactory separation of the compounds was obtained in less than 30 and 25 min, for the flavonoids and saponins, respectively. The methods were validated for linearity, repeatability, limits of detection (LOD) and limits of quantification (LOQ). Repeatability (inter- and intra-day, n = 6 and 9, respectively) showed less than 2% relative standard deviation (RSD). The LOD and LOQ were found to be 0.075 and 0.225 mg/mL, respectively, for vicenin-2 and 0.027 and 0.082 mg/100 mL, respectively, for mollugogenol B. The content of flavonoids and saponins of six single tablets was between 95 and 103% for flavonoids and 94-98% for saponins. The validated HPLC methods were employed to standardize a fingerprint of a laboratory produced purified extract, which could be used as a secondary standard for the routine quality control. Accordingly, the purified extract was found to contain 21.3% flavonoids (vicenin-2, 10%) and 25.4% saponins (glinuside G, 14.2%).

Apigenin↗

Extraction of heavy metals from MSW incinerator fly ash using saponins.

An extraction process with saponins was evaluated for removing heavy metals from MSW (municipal solid waste) incinerator fly ashes. Two different fly ashes, A and B, were treated on a laboratory scale with three triterpene-glycoside type of saponins, M, Q, and T, in the pH range 4-9. The results were compared with those of the HCI and EDTA treatment. The treatment with saponins extracted 20-45% of Cr from the fly ashes. Saponins were also effective in extracting Cu from fly ash A attaining 50-60% extraction. Saponin T extracted 100% of Pb from fly ash A at pH around 4. The extraction of Zn with the saponin treatment was similar to that of the HCl treatment. Further, Cr, Cu, Pb, and Zn were fractionated by sequential extraction to investigate the effect of saponins on each fraction. Extraction behavior of other elements during the saponin treatment was also studied. The leaching test on the residues received after the saponin treatment showed that the fly ashes were successfully detoxified to meet the landfilling guideline.

Air Pollutants↗

Effect of saponin on the transmucosal passage of beta-lactoglobulin across the proximal small intestine of normal and beta-lactoglobulin-sensitised rats.

The ability of saponins and glycoalkaloids to permeabilise the mammalian intestinal barrier has been previously demonstrated in vitro, leading to the hypothesis that membranolytic saponins may facilitate transfer to the tissues of otherwise excluded macromolecules. An enhanced uptake of, for instance, potentially allergenic species from the lumen is one of the factors that may affect the induction of food allergy, and its presentation in already sensitised individuals. In the experiments described here, an increase in the transmucosal uptake of the milk allergen beta-lactoglobulin (beta LG) was assessed in non-sensitised and sensitised Brown Norway rats in the presence of Gypsophila saponin. Isolated jejunal loops were exposed in vivo to either beta LG followed by saponin, saponin followed by beta LG or the two compounds simultaneously. Portal vein blood samples were collected and assayed for beta LG and rat mucosal mast cell protease (RCMP II) activity. Mucosal tissue was also examined histologically and assayed for histamine content. Sham-operated animals, exposed to physiological buffer alone, were included as controls and beta LG measurements corrected for this component which was negligible. No transfer of beta LG occurred in the absence of saponin in non-sensitised rats, whereas a significant enhancement was observed in the presence of saponin. beta LG was detected in the portal circulation of sensitised rats exposed to beta LG alone; however addition of saponin to the intestinal lumen further enhanced this uptake, possibly by an independent mechanism. Histological examination of the mucosal epithelium exposed to saponin revealed damage, especially at the villus tips. Mucosal histamine and serum RCMP II concentrations were consistent with the differences observed between sensitised and non-sensitised animals. It is concluded that exposure to food constituents capable of permeabilising the mucosal epithelium may increase the risk of sensitisation to dietary antigens.

Animals↗

Structure-function relationship among Quillaja saponins serving as excipients for nasal and ocular delivery of insulin.

The purpose of this investigation was to explore the structure-function relationship among naturally occurring Quillaja saponins and derivatives for their ability to stimulate insulin delivery from nosedrops and eyedrops and to test the hypothesis that stimulation of peptide drug delivery was correlated with surfactant strength. Native saponins, including QS-21, were purified from an aqueous extract of Quillaja saponaria bark by adsorption chromatography and HPLC. Native saponins were then deacylated by mild alkaline hydrolysis to form DS-1 and DS-2, derivatives that are smaller and more hydrophilic than their parent compounds. DS-1 was further treated either to reduce an aldehyde residue to form DS-1(R) or to remove the fucose-containing oligosaccharide to form QH-957. Rats receiving eyedrops or nosedrops formulated with insulin, but without any Quillaja saponins, showed no hypoglycemic response. Rats receiving eyedrops or nosedrops formulated with insulin plus saponins showed a dose-dependent hypoglycemic response, with the following rank order: QS-21 > DS-1 > DS-1(R) > DS-2 > QH-957. Surfactant strength was determined by measurement of the critical micellar concentration (cmc) and hemolysis of sheep erythrocytes. The cmc was lowest for the parent saponins QS-21 and QS-18, and increased for the deacylated saponin derivatives DS-1, DS-2, and QH-957; hemolysis of sheep erythrocytes was observed at low concentrations (approximately 0.006 mM) of the parent saponins, QS-21 and QS-18, at intermediate concentrations (0.06-0.08 mM) of DS-1 and DS-2, and at higher concentrations of DS-1(R) (0.45 mM) and QH-957 (1.5 mM). Hence, efficacy as an absorption-enhancing agent was greatest in those saponins with the lowest hemolytic titers and cmc values. However, this relationship was not a strict one, because DS-1, which differs from DS-2 only in the absence of one glucose residue, was significantly more potent than DS-2 in stimulating the absorption of insulin. DS-1 and DS-2 share a similar cmc and hemolytic titer, so this difference in efficacy must be due to some specificity beyond simple surfactant strength. Furthermore, DS-1 does not trigger an immune response when administered to animals, whereas QS-21 is a strong immune system activator. Therefore, DS-1 has emerged as an interesting candidate for inclusion in an eyedrop or nosedrop formulation.

Administration, Intranasal↗

Hemolysis of human erythrocytes with saponin affects the membrane structure.

Incubation of cells and tissues with saponin makes the lipid bilayer permeable to macromolecules. Ghosts (membrane preparations) of saponin-lysed erythrocytes do not reseal, thus indicating an irreversible damage of the lipid bilayer. We investigated the influence of disturbance of the lipid bilayer on membrane proteins by comparing ghosts of saponin-lysed erythrocytes with ghosts of cells lysed in hypotonic buffer. Transmission electron microscopy revealed destruction of the lipid bilayer and emergence of multilamellar buds in saponin-lysed ghosts. Freeze-fracture electron microscopy showed regions with crystalline lipids and an increase in particle-free areas on fracture faces. The number of protein sulfhydryl groups and the binding of hemoglobin were diminished in saponin-lysed ghosts. A Scatchard plot of hemoglobin binding revealed the decrease of high affinity binding sites. All these results indicate an aggregation of band 3 protein also demonstrated by laser scanning microscopy after incubation of cells labelled with eosin-5-maleimide with sublytic concentration of saponin. Hemolysis with saponin also affected the interaction between transmembrane proteins and the cytoskeleton. Dissociation of peripheral membrane proteins by incubation of ghosts in low salt buffer or by blocking sulfhydryl groups was increased and the association of spectrin with spectrin-depleted vesicles was decreased. The increased incorporation of the fluorescent probe Merocyanine 540 into saponin-lysed ghosts and the increased relative fluorescence quantum yield confirmed the perturbation of the lipid bilayer and the changed interaction between membrane lipids and intrinsic membrane proteins. Our results suggest that permeabilization of the lipid bilayer with saponin to admit the access of antibodies to the cytoplasmic surface of cells can aggregate transmembrane proteins and affect the immunocytochemical localization of associated proteins of the cytoskeleton.

Anion Exchange Protein 1, Erythrocyte↗

Protective effect of soybean saponins and major antioxidants against aflatoxin B1-induced mutagenicity and DNA-adduct formation.

Saponins from various plant sources have been suggested as possible anticarcinogens. Major dietary sources of saponins include legumes such as soybeans. This study was performed to determine the effect of soybean saponins on aflatoxin B(1)(AFB(1))-induced mutagenicity and AFB(1)-DNA adduct formation using Salmonella typhimurium and human liver hepatoma (HepG2) cells, respectively. Major antioxidants including L-ascorbic acid, alpha-tocopherol, all-trans-retinol, and butylated hydroxytoluene (BHT), previously reported to possess antimutagenic activity, were used as test materials to evaluate the relative effectiveness of saponins. Results indicated antimutagenicity was in the order of BHT > saponins > alpha-tocopherol > L-ascorbic acid. Soybean saponins exerted a significant effect, inhibiting the mutagenicity of AFB(1) by 52%, 64%, and 81% at concentrations of 600, 900, and 1,200 microg per plate, respectively. The amount of tritiated AFB(1) metabolites-DNA adducts formed in HepG2 cells was significantly reduced when cells were preincubated with 10 or 30 microg/ml of test materials. Soybean saponins inhibited AFB(1)-DNA adduct formation by 50.1% at a concentration of 30 microg/ml, whereas L-ascorbic acid and BHT reduced adduct formation by 38.4% and 32.6%, respectively, at the same concentrations. These results indicate that soybean saponins possess not only a significant antimutagenic activity but a strong inhibitory action against carcinogen-induced DNA damages. Soybean saponins possibly block the initiation stage of carcinogenesis, and further studies are required to elucidate the mechanisms of action.

Aflatoxin B1↗

Interactions between hemolytic saponins, bile salts and small intestinal mucosa in the rat.

The interaction between bile salts and saponins from Gypsophylla was investigated in vitro, using changes in transmural potential difference across isolated lengths of rat jejunum as an index of the capacity of the saponin to affect the permeability of intestinal mucosal cells. The addition of saponin (ca. 5 mM) to taurocholic acid in buffered saline (pH 7.4) led to a significant increase in viscosity, indicating the formation of a polymer. The viscosity of the solution was highest at low molar ratios of bile salt to saponin (0.5-2.0). Under these conditions there was an inhibition of the permeating effect of saponins on the gut, as judged by the rate of decline in transmural PD. There was no evidence of inflammation or functional damage to the jejunal mucosa of rats fed a diet containing Gypsophylla saponin (ca. 1.5% w/w) for 7 d but changes in villus morphology were observed. There was also evidence of an increased rate of mucosal cell proliferation. Serum cholesterol levels were significantly lower in saponin-fed rats than in controls, while the cholesterol content of the cecal contents was increased. These results suggest that despite the protective effect of bile salts, Gypsophylla saponins interact with the mucosa of the proximal small intestine in vivo, but at the dietary level used in this study the mucosa was protected by an enhanced rate of cell replacement. The loss of cholesterol via exfoliated mucosal cells may contribute to the hypocholesterolemic effect of saponins in rats.

Animals↗

Saponins as anticarcinogens.

Saponins, which are present in plants, have been suggested as possible anticarcinogens. They possess surface-active characteristics that are due to the amphiphilic nature of their chemical structure. The proposed mechanisms of anticarcinogenic properties of saponins include direct cytotoxicity, immune-modulatory effects, bile acid binding and normalization of carcinogen-induced cell proliferation. However, the anticarcinogenic effects of saponins from commonly consumed plant foods have not been studied. Soybeans are one of the most important sources of dietary saponins. They are the main protein supplier in many vegetarian diets. Our results showed that soybean saponins at the concentration of 150-600 ppm had a dose-dependent growth inhibitory effect on human carcinoma cells (HCT-15). Viability was also significantly reduced. Soybean saponins did not increase cell membrane permeability in a dose-dependent fashion, whereas gypsophilla saponin, a nondietary saponin, increased permeability with increasing concentrations. Electron microscopy indicated that soybean and gysophilla saponins alter cell morphology and interact with the cell membrane in different ways.

Antineoplastic Agents↗

Effects of a brief treatment with saponin on the contractile and electrical activities of isolated uterine muscle of pregnant rat.

The isolated longitudinal or circular muscle strip of rat uterus on day 20 of pregnancy becomes quiescent when incubated with Krebs solution for about 2 hr. Electric stimulation gave rise to a twitch-like contraction with a small amplitude instead of the spontaneous phasic contraction observed during the early period of incubation. When saponin (30-500 micrograms/ml) was applied, the basal tension was elevated and spontaneous contractions having large amplitude were elicited. The amplitude was larger than the phasic contraction of the K-contracture observed prior to the saponin treatment. The membrane was depolarized by about 25 mV, and periodic burst discharge was generated at the onset of saponin treatment. Raising the external Mg concentration from 0 to 2.4 mM in steps caused a depression of contractions in a dose-dependent manner before treatment with saponin, whereas the depression by Mg was much reduced after saponin treatment. The saponin-treated muscle recovered to exhibit control activity when tissues were superfused with Krebs solution for hours, i.e. the membrane was repolarized, the duration of action potential was detracted, and the depressant effect of Mg was again potentiated. For comparison, the effects of saponin and Mg on the contractions of ileal longitudinal muscle of pregnant rat were studied. In view of the above observations, the following were discussed. 1. Saponin impaired the depressant effect of Mg on the generation of contractions. 2. The membrane once impaired by saponin recovered in vitro probably under the influence of the genomic effect of ovarian hormones, a phenomenon which appeared characteristic for pregnant uterus.

Animals↗

Separation and characterization of saponins with adjuvant activity from Quillaja saponaria Molina cortex.

Saponins were purified from Quillaja saponaria Molina bark by silica and reverse phase chromatography. The resulting purified saponins were tested for adjuvant activity in mice. Several distinct saponins, designated QS-7, QS-17, QS-18, and QS-21, were demonstrated to boost antibody levels by 100-fold or more when used in mouse immunizations with the Ag BSA and beef liver cytochrome b5. These purified saponins increased titers in all major IgG subclasses. To determine optimal dose in mice for adjuvant response, QS-7 and QS-21 were tested in a dose-response study in intradermal immunization with BSA in mice; for both of these purified saponins, adjuvant response (determined by stimulation of ELISA titers to BSA) neared maximum at doses of 5 micrograms and was shown to plateau up to the highest dose tested, 80 micrograms. These purified saponins vary considerably in their toxicity, as assessed by lethality in mice; the main component, QS-18, being the most toxic. Saponins QS-7 and QS-21 showed no or very low toxicity in mice, respectively. None of these saponins stimulated production of reaginic antibodies. The monosaccharide composition of these saponins showed similar but distinct compositions with all four containing fucose, xylose, galactose and glucuronic acid. Predominant differences were observed in the quantities of rhamnose, arabinose, and glucose. Monomer m.w. (determined by size exclusion HPLC) were determined to range from 1800 to 2200.

Adjuvants, Immunologic↗

Quillaja saponin can modulate ovalbumin-induced IgE allergic responses through regulation of Th1/Th2 balance in a murine model.

Quillaja saponin is the extract from the balk of a South American tree, and it is considered to modulate immunological responses. We hypothesized that Quillaja saponin may change allergy-associated cytokine profile and antigen-specific immune responses. The purpose of this study is to investigate whether Quillaja saponin can suppress ovalbumin (OVA)-induced IgE-mediated allergic responses through promoting a dominant Th1 immune response. The spleen cells from BALB/c mice, which were primed by OVA, were used for an in vitro challenge test. The level of total and OVA-specific IgE, IL-4, IFN-gamma, and IL-12 was determined by enzyme-linked immunosorbent assay (ELISA). BALB/c mice were orally administered with saponin for 35 days. The mice were immunized intraperitoneally with OVA on days 14 and 21. After intraperitoneal challenge with OVA on day 35, anaphylactic symptoms were monitored. Total and specific IgE and IgG, specific IgG1 and IgG2a, and histamine levels in serum were analyzed by ELISA. The increase of IL-12 and IFN-gamma levels was observed in the presence of Quillaja saponin, while the IL-4 level was decreased. Furthermore, Quillaja saponin suppressed total and OVA-specific IgE secretion in spleen cells. Balb/c mice that were orally administered Quillaja saponin exhibited lower total and OVA-specific IgE and OVA-specific IgG secretions, whereas total IgG levels remained unchanged. Suppression of OVA-specific IgG1 and an increase of OVA-IgG2a were observed in mice fed saponin. Quillaja saponin also decreased serum histamine levels and diminished anaphylactic symptoms. The present study indicates that Quillajasaponin can suppress allergen-specific IgE-mediated reactivity in a murine model of food allergy, which results from shifting from a Th2-dominated to a Th1-dominated immune response.

Animals↗