sitosterol--(3beta)-isomer and gamma-sitosterol

The root bark of Strychnos innocua has long been employed by traditional healers to treat a variety of ill health conditions including fungal infections. The objective of this study was to isolate, characterized and evaluate the antifungal potential (insilico approach) of some steroids from root bark of S. innocua. Conventional method of column chromatography was carried out on the ethyl acetate root bark extract of S. innocua, leading to the isolation of two steroids. On the basis of 1D NMR, 2D NMR, GC-ESI/MS analyses, and literature comparisons, the compounds were characterized as Stigmast-5-en-3β-ol (1) and Campest-5-en-3β-ol (2). This is the first time these compounds have been isolated from the plant. The results of the in silico modelling of the compounds 1, 2, and fluconazole (control drug) with the binding sites of SAP2 from Candida albicans (PDB: 1EAG) demonstrated that the binding energies were -8.3, -8.0, and -7.1 kcal/mol, respectively. However, the modelling with binding sites of a deglycating enzyme fructosamine oxidase from Aspergillus fumigatus (PDB: 3DJE) demonstrated that the binding energies were -5.9, -7.2, and -8.0 for Stigmast-5-en-3β-ol (1) and Campest-5-en-3β-ol (2), and fluconazole, respectively. In conclusion, the study found that Stigmast-5-en-3-ol and Campest-5-en-3-ol are both present in the root bark of S. innocua. The compounds exhibited promising interaction with the binding sites of the protein target (SAP2 from C. albicans) compare to fluconazole.

Topics: Antifungal Agents; Computer Simulation; Fluconazole; Plant Bark; Plant Roots; Steroids; Strychnos

Topics: Acanthaceae; Anti-Bacterial Agents; Dose-Response Relationship, Drug; Microbial Sensitivity Tests; Molecular Structure; Plant Leaves; Sitosterols; Staphylococcus aureus; Structure-Activity Relationship

Beta-sitosterol (Sit) widely exists in natural plants, is classed as phytosterol and known as the "key of life". Most pharmacological studies and clinical applications are limited because of the fact that Sit is difficult to be solved. Therefore, it is viable to enhance pharmacologic activities of Sit by using its derivatives which can be obtained through the modification of Sit. In this study, 4 kinds of new Sit derivatives were obtained by the esterification reaction. Further, the hepatoprotective effects of Sit and its derivatives were investigated against acute liver injury induced by lipopolysaccharide/d-galactosamine (LPS/GalN) in mice and its mechanism was illustrated by western blot analysis and real-time PCR. The results demonstrated that among its derivatives, 2-naphthoyl Sit ester (Sit-N) (50 mg/kg) showed the strongest activities against acute liver injury. Final experimental results showed that Sit-N significantly decreased the serum activity of aspartate transaminase (AST) and alanine aminotransferase (ALT); Sit-N also markedly reduced tumor necrosis factor (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) levels. Meanwhile, Sit-N drastically improved the activities of antioxidant enzymes such as superoxide dismutase (SOD), glutathione (GSH) and catalase (CAT), and suppressed the expression of malondialdehyde (MDA). Results also displayed that over-expression of Toll like receptor 4 (TLR4) and nuclear factor-kappa B (NF-κB) induced by LPS/Gal N were inhibited by Sit-N. Meanwhile, the expression of nuclear respiratory factor2 (Nrf2) and heme oxygenase-1 (HO-1) were enhanced. The results all above verified the effectiveness of Sit-N against acute liver injury induced by LPS/GalN mediated by TLR4 and Nrf2 pathways.

Topics: Animals; Anti-Inflammatory Agents; Cell Survival; Chemical and Drug Induced Liver Injury; Dose-Response Relationship, Drug; Galactosamine; Inflammation; Lipopolysaccharides; Mice; Molecular Structure; Oxidation-Reduction; Sitosterols; Structure-Activity Relationship

Inhibitors of thrombin, a key enzyme in the blood coagulation cascade, are of great interest because of their selective specificity and effectiveness in anticoagulation therapy against cardiovascular disorders. The natural soybean phytosterol, β-sitosterol (BSS) demonstrated anticoagulant activity by dose-dependent inhibition of thrombin in an uncompetitive manner with a K

Topics: Animals; Anticoagulants; Antithrombins; Catalysis; Disease Models, Animal; Female; Fibrinogen; Humans; Male; Mice; Plants; Platelet Aggregation Inhibitors; Sitosterols; Thrombin; Thrombosis

γ-sitosterol isolated from Lippia nodiflora was taken as ligand for molecular docking. The molecular targets, glucokinase, Fructose 1, 6- bisphosphatase 1, Human multidrug resistance protein 1 and Cytochromes P450 whose crystallographic structures are available on the PDB database as 1V4S, 2JJK, 3LC4, 2CBZ respectively, were used for the docking analysis using the Autodock tool v 4.2 and ADT v1.5.4 programs. The docking studies of the ligand γ- sitosterol with four different target proteins showed that this is a good molecule which docks well with various targets related to diabetes mellitus. Hence γ-sitosterol can be considered for developing into a potent antidiabetic drug.

Topics: Computational Biology; Diabetes Mellitus; Humans; Hypoglycemic Agents; Models, Molecular; Protein Conformation; Sitosterols

In our preliminary screening study on the anti-inflammatory activity, eight triterpenes, one sterol, and one chalcone were isolated from the CH(2)Cl(2)-soluble extract of the stems and leaves of Rhus sylvestris Siebold and Zucc (Anacardiaceae). On the basis of their spectroscopic data, these compounds were identified as 10alpha-cucurbitadienol (1), glut-5-en-3-ol (2), beta-amyrin acetate (3), beta-amyrin (4) and lupeol (5), cycloart-24-en-3-one (6), cycloart-25-en-3,24-dione (7), 24-hydroxycycloart-25-en-3-one (8), beta-sitosterol (9), and 2'-hydroxy-4,4'-dimethoxychalcone (10). All of them were isolated from this plant for the first. Furthermore, the compounds in non-cytotoxic concentrations (0-1.0microM) were tested for their ability to block inflammatory cytokine secretion in the presence of LPS in the murine RAW264.7 macrophage cell line. Among the compounds that were tested, compounds 8 and 9 reduced the LPS-induced secretion of IL-6, as well as TNF-alpha, in a mouse RAW264.7 macrophage cell line. Moreover, compounds 2, 3, 7, and 10 specifically diminished only the secretion of TNF-alpha even in 0.01microM concentrations. It is thus suggested that they are potential therapeutics of TNF-alpha-related diseases and conditions, such as transplant rejection, type II diabetes, and atherosclerosis.

Topics: Anacardiaceae; Animals; Anti-Inflammatory Agents; Cell Line; Cytokines; Humans; Interleukin-6; Lipopolysaccharides; Macrophages; Mice; Plant Leaves; Sitosterols; Triterpenes; Tumor Necrosis Factor-alpha

Recently several plant derived natural compounds have been screened for their anticancer activity in order to identify putative compounds with novel structures or mechanism of action. In the present study, fruits of Poncirus trifoliata were extracted with acetone and loaded onto silica gel column chromatography. The column was eluted with different solvents to obtain two bioactive compounds. The purity of compounds was analyzed by HPLC and their structures were identified by 1H and 13C NMR experiments as beta-sitosterol and 2-hydroxy-1,2,3-propanetricarboxylic acid 2-methyl ester (HPCME). beta-Sitosterol, HPCME, and trolox were tested for their antioxidant capacity by oxygen radical absorbance capacity (ORAC) measurement. Further, these compounds were tested for their inhibition of cancer cell proliferation and apoptosis using human colon cancer cell line (HT-29). These results were compared with the corresponding activity on non-cancerous (COS-1 fibroblast) cells. Cell proliferation and induction of apoptosis were determined by MTT assay and nuclear staining. The MTT assay indicated that both the compounds exhibited differential inhibition at various concentrations. Significant arrest of cell growth was observed with beta-sitosterol even at low concentration such as 0.63 microM in 48 h and none of the compounds exerted any apparent cytostatic effects on the non-cancerous COS-1 fibroblast cells. Growth inhibition assay suggested the potential use of bioactive compounds as cancer chemopreventive and therapeutic agents. This is the first report on HPCME isolation and identification from Rutaceae family and beta-sitosterol from P. trifoliata.

Topics: Animals; Antioxidants; Carboxylic Acids; Cell Extracts; Cell Line; Cell Proliferation; Chlorocebus aethiops; Chromatography, High Pressure Liquid; Colonic Neoplasms; Esters; Humans; Magnetic Resonance Spectroscopy; Molecular Structure; Poncirus; Sitosterols

Two ATP-binding cassette (ABC) proteins, ABCG5 and ABCG8, have recently been associated with the accumulation of dietary cholesterol in the sterol storage disease sitosterolemia. These two 'half-transporters' are assumed to dimerize to form the complete sitosterol transporter which reduces the absorption of sitosterol and related molecules in the intestine by pumping them back into the lumen. Although mutations altering ABCG5 and ABCG8 are found in affected patients, no functional demonstration of sitosterol transport has been achieved. In this study, we investigated whether other ABC transporters implicated in lipid movement and expressed in tissues with a role in sterol synthesis and absorption, might also be involved in sitosterol transport. Transport by the multidrug resistance P-glycoprotein (P-gp; Abcb1), the multidrug resistance-associated protein (Mrp1; Abcc1), the breast cancer resistance protein (Bcrp; Abcg2) and the bile salt export pump (Bsep; Abcb11) was assessed using several assays. Unexpectedly, none of the candidate proteins mediated significant sitosterol transport. This has implications for the pathology of sitosterolemia. In addition, the data suggest that otherwise broad-specific ABC transporters have acquired specificity to exclude sitosterol and related sterols like cholesterol presumably because the abundance of cholesterol in the membrane would interfere with their action; in consequence, specific transporters have evolved to handle these sterols.

Topics: 3T3 Cells; Animals; ATP Binding Cassette Transporter, Subfamily B, Member 1; ATP Binding Cassette Transporter, Subfamily G, Member 2; ATP-Binding Cassette Transporters; Biological Transport; Flow Cytometry; Mice; Mice, Knockout; Microscopy, Confocal; Multidrug Resistance-Associated Proteins; Neoplasm Proteins; Sitosterols; Spodoptera