Reduction of blood pressure by soybean saponins, renin inhibitors from soybean, in spontaneously hypertensive rats

The effect of commercial purified soybean saponin on renin activity and blood pressure was investigated. Soybean saponin significantly inhibited human renin in vitro with IC(50)=59.9 μg/ml. Orally administered soybean saponin at 80 mg/kg of body weight per day to spontaneously hypertensive rats for 8 weeks significantly decreased the blood pressure.     

Reduction of Blood Pressure by Soybean Saponins,Renin Inhibitors from Soybean,in Spontaneously Hypertensive Rats

The effect of commercial purified soybean saponin on renin activity and blood pressure was investigated. Soybean saponin significantly inhibited human renin in vitro with IC₅₀=59.9 μg/ml. Orally administered soybean saponin at 80 mg/kg of body weight per day to spontaneously hypertensive rats for 8 weeks significantly decreased the blood pressure.     

无患子水提皂素液发酵精制联产生物表面活性剂槐糖脂

无患子水提皂素液,经纤维二糖酶水解,以无患子水提水解液为底物,接种丘陵假丝酵母,将水提液中糖组分发酵转化为槐糖脂,得到天然皂素及生物表面活性剂复合产物。在发酵过程中,2%的丘陵假丝酵母菌种接种量,溶液中葡萄糖消耗速率最快;在水提水解液中额外添加大豆油作为补充碳源能较大幅度降低溶液表面张力。经过发酵转化,溶液中表面活性物质浓度达到52.48 g/L,比发酵前提高了23.4%,溶液表面张力值明显降低。无患子精制发酵液中不含糖类成分,是理想的液体洗涤剂生产原料。     

Release of soluble resident as well as secretory proteins from HepG2 cells by partial permeabilization of rough-endoplasmic-reticulum membranes.

Secretory proteins migrate from the rough endoplasmic reticulum (ER) to the Golgi complex at different rates. Selective retention of specific proteins to rough-ER membrane constituents could explain this phenomenon. We have permeabilized HepG2 cells with low concentrations of saponin. Release of newly synthesized proteins was studied after brief labelling in the presence of [35S]methionine. The efflux of several secretory proteins was studied at various saponin concentrations; a 2-fold higher saponin concentration was required to release transferrin compared with that required to release albumin and orosomucoid. Glucosidase II, a soluble resident protein of the ER, is released at the same saponin concentration as albumin. Saponin did not destroy the membrane skeleton structure; at the concentrations used, the integral membrane protein G of vesicular-stomatitis virus remained fully associated with the cells.     

A steryl glycoside fraction from Momordica charantia seeds with an inhibitory action on lipid metabolism in vitro

A saponin fraction was isolated from Momordica charantia seeds by delipidation, saline extraction, ammonium sulfate precipitation, and extraction of the resulting supernatant with n-butanol. Thin-layer chromatography, in the upper phase of the n-butanol--ethyl acetate--water (4:1:5, by volume) system on plastic sheets coated with silica gel 60 F254, revealed the presence of a single spot after spraying with 10% sulfuric acid. The lack of contamination of the saponin preparation with proteins was judged by the absence of protein bands in sodium dodecyl sulfate--polyacrylamide gel electrophoresis, agarose electrophoresis and agarose diffusion, and by the absence of an absorption maximum around 278 nm. The saponin acted as a noncompetitive inhibitor of corticotropin, glucagon, and epinephrine in lipolysis in isolated rat adipocytes, and it also antagonized dibutyryl cAMP induced lipolysis. The antilipolytic activity was resistant to heat, trypsin, chymotrypsin, pronase, and glutathione, in keeping with the chemical nature of saponin. Incorporation of [3-3H]glucose into lipid was inhibited. Adipocyte viability and ATP content were not affected by the saponin, suggesting that its inhibitory effects on lipolysis and lipogenesis were not due to an adverse effect on cell viability.     

Real-time analysis of membrane permeabilizing effects of oleanane saponins

Saponins are a group of plant and marine derived glycosides with numerous biological functions. Two important characteristics of certain plant saponins are their ability to enhance cytotoxicity of type I ribosome inactivating proteins and stimulation of the immune system. The main objective of the present study was to investigate in real-time the permeabilizing effects of saponins on cell membrane. A set of oleanane saponins (glycyrrhizinic acid, Gypsophila, Saponaria and Quillaja saponins) and a steroid saponin (digitonin) were tested. The effects of these saponins on lysosomal membranes and hemolysis, along with their charge were also studied. Real-time monitoring of cell membrane permeabilization facilitated a highly sensitive analysis of the cellular kinetics. Saponins showed variable permeabilizing effects on cellular and lysosomal membranes at concentrations from 6 μM and hemolysis from 3 μM. Further, the results suggest that charge of the saponin may be relevant for permeabilizing effects of oleanane saponins.     

食物单宁和皂苷对小白鼠食物选择的影响

通过饲喂自然种子及人工饲料块等单宁和皂苷含量不同的食物,研究了食物中单宁或皂苷水平对小白鼠(Mus musculus domesticus)食物选择的影响,以及取食经历在其食物选择中的作用。结果表明,(1)无论有无取食经历,小白鼠均优先取食次生物质含量低的锥栗(Castanea henryi)或花生(Arachis hypogaea),而很少取食单宁含量高的栓皮栎(Quercus variabilis)或皂苷含量高的油茶(Camelia oleifera);(2)小白鼠的取食经历能增强其对种子中单宁或皂苷水平的识别,从而减少其摄入;(3)小白鼠的食物摄入量随单宁或皂苷含量的增加而显著降低。本研究说明单宁或皂苷均可显著影响动物的食物选择,且取食经历能增强动物对食物的识别能力。     

Inhibition of increases in blood glucose and serum neutral fat by Momordica charantia saponin fraction

Focusing on a functional component of Momordica charantia, saponin, we investigated its effects on serum glucose and neutral fat levels. Saponin was extracted as a butanol-soluble fraction (saponin fraction) from hot blast-dried Momordica charantia powder. The disaccharidase-inhibitory activity and the pancreatic lipase-inhibitory activity of the saponin fraction were measured, and in vivo sugar- and lipid-loading tests were performed. The saponin fraction inhibited disaccharidase activity and elevation of the blood glucose level after sucrose loading. The fraction also markedly inhibited pancreatic lipase activity and elevation of the serum neutral fat level after corn oil loading. Based on these findings, the main active component related to the anti-diabetic effect of Momordica charantia is present in the butanol fraction, and it may be saponin. The blood glucose and serum neutral fat-lowering effects of Momordica charantia were closely associated with its inhibitory activity against disaccharidase and pancreatic lipase.     

西洋参悬浮细胞发酵工艺研究

探讨了西洋参悬浮细胞分步培养与稀土、D-半乳糖和甘露醇等诱导子对悬浮细胞生长及皂甙产量的影响。发现继代4d后换液一次再继续培养获得的培养物,在皂甙产率和糖利用率等方面优于连续培养;D-半乳糖作为诱导子,对悬浮培养的西洋参细胞生长、皂甙产率及皂甙的分泌等方面都有非常明显的促进作用。     

Saponin effects of prolactin-like stimulation of ornithine decarboxylase activity in mouse mammary gland explants.

Saponin, a naturally occurring plant glycoside, was found to elicit a prolactin-like stimulation of ornithine decarboxylase (ODC) activity in mouse mammary gland explants. A dose-response activation of ODC was observed with saponin at concentrations between 2 and 10 micrograms/ml. At concentrations of 10 and 15 micrograms/ml, saponin effected a response similar to that of PRL; when tested in concert, PRL and saponin caused a nonadditive response. The time-course of the saponin and PRL effects on ODC activation were not different; a maximum response occurred 2-4 hours after addition of saponin. The saponin and PRL responses were abolished by antibiotics (puromycin and cyclohexamide) that inhibit protein synthesis, but not by actinomycin D which inhibits RNA synthesis. Finally, saponin, by itself, did not affect the rate of milk product formation, but at higher concentrations (above 0.5 microgram/ml) impaired the PRL stimulation of lipid and casein synthesis.     

Detergent-insoluble glycosphingolipid/cholesterol microdomains of the myelin membrane

Glycosphingolipids and cholesterol form lateral assemblies, or lipid 'rafts', within biological membranes. Lipid rafts are routinely studied biochemically as low-density, detergent-insoluble complexes (in non-ionic detergents at 4 degrees C; DIGs, detergent-insoluble glycosphingolipid/cholesterol microdomains). Recent discrepancies recommended a re-evaluation of the conditions used for the biochemical analysis of lipid rafts. We have investigated the detergent insolubility of several known proteins present in the glycosphingolipid/cholesterol-rich myelin membrane, using four detergents representing different chemical classes (TX-100, CHAPS, Brij 96 and TX-102), under four conditions: detergent extraction of myelin either at (i) 4 degrees C or (ii) 37 degrees C, or at 4 degrees C after pre-extraction with (iii) saponin or (iv) methyl-beta-cyclodextrin (MbetaCD). Each detergent was different in its ability to solubilize myelin proteins and in the density of the DIGs produced. Brij 96 DIGs floated to a lower density than other detergents tested, possibly representing a subpopulation of DIGs in myelin. DIGs pre-extracted with saponin were denser than DIGs pre-extracted with MbetaCD. Furthermore, pre-extraction with MbetaCD solubilized proteolipid protein (known to associate with cholesterol), whereas pre-extraction with saponin did not, suggesting that saponin is less effective as a cholesterol-perturbing agent than is MbetaCD. These results demonstrate that DIGs isolated by different detergents are not necessarily comparable, and that these detergent-specific DIGs may represent distinct biochemical, and possibly physiological, entities based on the solubilities of specific lipids/proteins in each type of detergent.     

Effects of pulsed magnetic field treatment of soybean seeds on calli growth,cell damage,and biochemical changes under salt stress

The effects of magnetic field (MF) treatments of soybean seeds on calli growth, cell damage, and biochemical changes under salt stress were investigated under controlled conditions. Soybean seeds were exposed to a 1.0 Hz sinusoidal uniform pulsed magnetic field (PMF) of 1.5 µT for 5 h/day for 20 days. Non‐treated seeds were considered as controls. For callus regeneration, the embryonic axis explants were taken from seeds and inoculated in a saline medium with a concentration of 10 mM NaCl for calli growth analysis and biochemical changes. The combined treatment of MF and salt stress was found to significantly increase calli fresh weight, total soluble sugar, total protein, and total phenol contents, but it decreased the ascorbic acid, lipid peroxidation, and catalase activity of calli from magnetically exposed seeds compared to the control calli. PMF treatment significantly improved calli tolerance to salt stress in terms of an increase in flavonoid, flavone, flavonole, alkaloid, saponin, total polyphenol, genistein, and daidzein contents under salt stress. The results suggest that PMF treatment of soybean seeds has the potential to counteract the adverse effects of salt stress on calli growth by improving primary and secondary metabolites under salt stress conditions. Bioelectromagnetics 33:670–681, 2012. © 2012 Wiley Periodicals, Inc.     

Manipulation of saponin biosynthesis by RNA interference-mediated silencing of ��-amyrin synthase gene expression in soybean

Soybean seeds contain substantial amount of diverse triterpenoid saponins that influence the seed quality, although little is known about the physiologic functions of saponins in plants. We now describe the modification of saponin biosynthesis by RNA interference (RNAi)-mediated gene silencing targeted to β-amyrin synthase, a key enzyme in the synthesis of a common aglycon of soybean saponins. We identified two putative β-amyrin synthase genes in soybean that manifested distinct expression patterns with regard to developmental stage and tissue specificity. Given that one of these genes, GmBAS1, was expressed at a much higher level than the other (GmBAS2) in various tissues including the developing seeds, we constructed two RNAi vectors that encode self-complementary hairpin RNAs corresponding to the distinct regions of GmBAS1 under the control of a seed-specific promoter derived from the soybean gene for the α′ subunit of the seed storage protein β-conglycinin. These vectors were introduced independently into soybean. Six independent transgenic lines exhibited a stable reduction in seed saponin content, with the extent of saponin deficiency correlating with the β-amyrin synthase mRNA depletion. Although some transgenic lines produced seeds almost devoid of saponins, no abnormality in their growth was apparent and the antioxidant activity of their seeds was similar to that of control seeds. These results suggest that saponins are not required for seed development and survival, and that soybean seeds may therefore be amenable to the modification of triterpenoid saponin content and composition through molecular biologic approaches.     

Elastin-like polypeptide and γ-zein fusions significantly increase recombinant protein accumulation in soybean seeds

Transgenic Research - Soybean seeds are an ideal host for the production of recombinant proteins because of their high content of proteins, long-term stability of seed proteins under ambient...     

Modified cell ELISA to determine the solubilization of cell surface proteins: Applications in GPI-anchored protein purification

A major step in purifying membrane bound proteins involves the solubilization of the protein of interest from the cell membranes. Glycosylphosphatidyl inositol (GPI)-anchored proteins pose a singular problem in this solubilization step since they are found in detergent-resistant membrane complexes and accordingly are insoluble in cold Triton X-100. In this study we have developed a modified cell ELISA that determines the solubility of these cell surface proteins under various solubilization conditions. Using this non-radioactive method we show that the combination of saponin/Triton X-100 at 4 degrees C solubilized GPI-anchored proteins more efficiently than Triton X-100 at 4 degrees C. The combination of saponin/Triton X-100 at 4 degrees C avoids the potential of activating proteases that occurs when using Triton X-100 at 37 degrees C. Furthermore, our method also shows the saponin/Triton X-100 solubilized GPI-anchored proteins equivalent to the more expensive octyl beta-glucoside. This is a particularly important consideration in large-scale protein purification. This method obviates the need to use radioactivity, gel electrophoresis and immunoblotting procedures. The solubilization conditions determined by this modified ELISA are readily translated to the practical application of large-scale protein purification as demonstrated in the purification of two different recombinant GPI-anchored proteins, GPI-hB7-1 (CD80) and GPI-mICAM-1 (CD54).     

茶皂素对菜青虫的拒食作用方式及机制

采用注射、熏蒸、点滴、饲喂、内吸等方法测定了茶皂素对菜粉蝶Ｐｉｅｒｉｓ　ｒａｐａｅ　Ｌ．５龄幼虫的拒食活性。结果表明,茶皂素对５龄菜青虫具有很强的饲喂拒食活性,２４小时选择性拒食中浓度ＡＦＣ_(５０)为４０．８８ｍｇ／Ｌ,非选择性拒食中浓度为３２１．９２ｍｇ／Ｌ;并具有一定的内吸拒食作用,尤其是根部施药法内吸拒食活性较明显。茶皂素对菜青虫无显著的注射、熏蒸、点滴拒食作用。     

Effects of a series of alcohols on the binding of a fluorescent dye to erythrocyte membranes.

1. The effects of a series of aliphatic alcohols (methanol to octanol) on membrane proteins of erythrocytes were studied by monitoring the flueorescence of a dye (1-anilino-8-naphthalenesulfonic acid (ANS)) that adsorbs to erythrocyte ghost membranes. Low concentrations of all the alcohols reduced the ANS fluorescence of the membrane-ANS suspensions; lent to those which protect against hypotonic hemolysis on intact erythrocytes; higher concentrations markedly increased the fluorescence. Ethanol and methanol decreased ANS fluorescence at all concentrations. 2. Lytic concentrations of saponin did not increase ANS fluorescence and did not modify the membrane action of the alcohols. 3. None of these effects were observed in liposomes prepared from lipid extracts of the erythrocyte membrane. 4. Since the apparent dissociation constant for the ANS-membrane interaction was unchanged in the presence of the alcohols, it was assumed that the fluorescence changes anesthetic concentration of the alcohols alter the conformation of membrane proteins, as indicated by the decreased number of ANS binding sites.     

Tannins and saponin: Interaction in herbivore diets

Mice allowed to choose between diets containing tannin or saponin did not experience food intake depression, weight loss or high faeces weight to food weight ratios. Diets containing tannin produced all these effects and saponin diets resulted in weight losses and high faeces to food ratios. Mice provided with diets containing both tannin and saponin in predetermined proportions experienced weight losses similar to, or greater than, those of mice fed diets containing either toxin alone. Urinary glucuronide production by mice provided with a choice of tannin and saponin diets was less than that of mice feeding on diets containing either tannin or saponin alone. Simultaneous consumption of tannin and saponin (in the right proportions) may promote chemical interactions that inhibit the toxins' absorption from the intestinal tract. This type of interaction is likely to have influenced the evolution of herbivore feeding behaviour.     

Effects of diquat on pigment-protein complexes of thylakoid membranes in soybean and maize plants

Soybean (Glycine max Merrill) and maize (Zea mays L.) plants were exposed for 5 to 48 h to the herbicide diquat under "white light" (WL) or far-red radiation (FR) (photon fluence rate of 30 μmol m^-2 s^-1). The WL enhanced diquat effect on chlorophyll content in soybean plants, while FR had the same effects on maize plants. After 5 h, diquat increased the content of polypeptides bound to light-harvesting proteins in both plants. This revised version was published online in July 2006 with corrections to the Cover Date.