Stigmasterol Research Articles

Terpenes from Otostegia integrifolia

The essential oil and chloroform extract of air-dried leaves of Otostegia integrifolia Benth. were investigated for the first time using analytical and preparative gas chromatography (GC), GC-mass spectrometry (MS) and NMR techniques. A total of 40 constituents including monoterpenes, sesquiterpenes, diterpenes and their derivatives were identified. A prenylbisabolane type diterpene, 1-methyl-4-(5, 9-dimethyl-1-methylene-deca-4, 8- dienyl) cyclohexene was identified as a major component. The chloroform extract of the leaves yielded two new labdane type diterpenoids, 15, 16-epoxy-3a, 9a-dihydroxy-labda- 13(16), 14-diene and 9(13), 15(16)-diepoxy-3a-hydroxy-16-dihydrolabda-14-ene, a saturated hydrocarbon, pentatriacontane, and stigmasterol. The structures of the isolated compounds were established by spectroscopic methods. Keywords: Otostegia integrifolia Benth.; Lamiaceae; Essential oil, Furanolabdane diterpenes, Otostegindiol, Preotostegindiol, (+)-Axinyssene, Pentatriacontane, Stigmasterol

Stigmasterol activates Cdc42-Arp2 and Erk1/2-Creb pathways to enrich glutamatergic synapses in cultures of brain neurons

The naturally occurring phytosterol stigmasterol (ST) ameliorates scopolamine-induced memory impairment and promotes neuritogenesis and synaptogenesis. Therefore, in this study, we investigated the hypothesis that ST promotes spinogenesis and the formation of glutamatergic synapses and thus improves memory in cultured rat hippocampal neurons. Immunoblots showed that ST activated extracellular signal–regulated kinase 1/2 (Erk1/2) and cAMP response element binding protein (Creb), and upregulated actin-related protein 2 (Arp2) and cell division cycle 42 (Cdc42), central components in actin organization and spine head development. Consistently, DiO staining of live neurons showed that ST increased the densities of dendritic filopodia and mushroom-type mature spines, and immunocytochemistry showed an increase in the expressions of the GluN2A and GluN2B subunits of N-methyl-D-aspartate receptors (NMDARs) at synapses. Finally, staining the recycling membranes with FM1-43 showed that ST increased the sizes of synaptic vesicle pools at presynaptic terminals. Together, our data indicate that ST upregulates Cdc42‐Arp2/3 and Erk1/2-Creb signaling and thus induces the formations of mushroom-type spines and glutamatergic excitatory synapses.

Huberine, a New Canthin-6-One Alkaloid from the Bark of Picrolemma huberi.

A new alkaloid, Canthin-6-one, Huberine (1), together with three known compounds including 1-Hydroxy-canthin-6-one (2), Canthin-6-one (3) and stigma sterol (4), were isolated from the stem bark of Picrolemma huberi. The isolation was achieved by chromatographic techniques and the purification was performed on a C18 column using acetonitrile/water (90:10, v/v) with 0.1% formic acid as the mobile phase. The structural elucidation was performed via spectroscopic methods, notably 1D- and 2D-NMR, UV, IR, MS and HRMS. The antiplasmodial activity of the compounds was studied.

Stigmasterol upregulates immediate early genes and promotes neuronal cytoarchitecture in primary hippocampal neurons as revealed by transcriptome analysis

BackgroundThe hippocampus is a vulnerable brain region that is implicated in learning and memory impairment by two pathophysiological features, that is, neurite regression and synaptic dysfunction, and stigmasterol (ST), a cholesterol-equivalent phytosterol, is known to facilitate neuromodulatory effects. PurposeTo investigate the neuromodulatory effects of ST on the development of central nervous system neurons and the molecular bases of these effects in primary hippocampal neurons. MethodsRat embryonic (E18-19) brain neurons were cultured in the absence or presence of ST (75 µM). Neuritogenic activities of ST were evident by increases in various morphometric parameters. To identify underlying affected genes, total RNA was isolated on day in vitro 12 (DIV 12) and mRNA high throughput sequencing (mRNA-Seq) was performed. Affected key genes for neuronal development were identified using bioinformatics tools and their upregulations were confirmed by immunocytochemistry. ResultsAmong the differentially expressed 17,337 RefSeq genes, 445 genes (up/down 293/157) passed the p-value < 0.05 criterion, 52 genes (up/down; 37/13) had a p-value < 0.05 and a false discovery rate (FDR) q-value of < 0.2, and 24 genes (up/down; 20/4) passed the more stringent criterion of both p < 0.05 and q < 0.05. After applying a stringent FDR q-value cutoff of < 0.2, it was found ST induced many immediate early genes (IEGs), and that a major proportion of upregulated genes were related to central nervous system (CNS) development (neurite outgrowth or synaptic transmission). In a Venn diagram for CNS development Gene Ontologies (GOs) (i.e., axon development, dendrite development, modulation of synaptic transmission), Reln emerged as a central player in these processes, and highly interconnected ‘hub’ genes, including Dcx, Egr1, Ntrk2, and Slc24a2, were revealed by gene co-expression networks. Finally, transcriptomic data was confirmed by immunocytochemistry of primary hippocampal neurons. ConclusionThe study indicates that ST upregulates genes for neuritogenesis and synaptogenesis, and suggests ST be viewed as a potential resource for improving brain functions.

Determination of sterols in tobacco leaves by gas chromatography-tandem mass spectrometry

The purpose of the study was to establish an analytical method to simultaneously determine multiple sterols in tobacco leaves rapidly and accurately. In this gas chromatography-triple quadruple tandem mass spectrometry (GC-MS/MS) based method, various conditions for sterols extraction were assessed and the instrumental operation parameters were optimized with 5α-cholane as an internal standard. Using this method, eight plant sterols, namely lanosterol, campesterol, chenodeoxycholic acid, cholesterol, β-sitosterol, stigmasterol, dihydrotachysterol and ergosterol, were separated in less than 30 min. The linear correlation coefficients were over 0.998 9, the low detection limits were in the range of 0.010 3-0.141 4 μg/g. The recoveries were from 87.30% to 115.60%, with low standard deviations. In conclusion, this method demonstrates good repeatability, accuracy, and high sensitivity, and is best suited for rapid analysis of multiple sterols in tobacco leaves.

Separation of ‎STIGMA STEROL using magnetic molecularly imprinted nanopolymer fabricated by sol-gel method

Separation of ‎STIGMA STEROL using magnetic molecularly imprinted nanopolymer fabricated by sol-gel method

Determination of oleanolic acid, ursolic acid, lupeol, and stigmasterol by high-performance thin-layer chromatographic method in Oldenlandia corymbosa Linn.

Oldenlandia corymbosa Linn. (Rubiaceae) is an important herb traditionally used as a febrifuge and liver tonic. In this study, a high-performance thin-layer chromatography (HPTLC) method has been established for the quantification of four bioactive markers, oleanolic acid (OA), ursolic acid (UA), lupeol (LU), and stigmasterol (ST), in the whole plant of O. corymbosa. Separation was achieved on silica gel 60 F254 HPTLC plates using hexane-ethyl acetate-methanol (8.2:1.8:0.5, v/v) for oleanolic acid and ursolic acid; and toluene-methanol (9.4:0.6, v/v) for lupeol and stigmasterol as the mobile phases. The quantitation of the four markers was carried out using the densitometric scanning at 540 nm after derivatization using sulfuric acid reagent. The linear regression analysis data for the calibration plots showed a good linear relationship (r2 = 0.9831–0.9979) in the concentration range of 1200–4200 ng for oleanolic acid, 400–1400 ng for ursolic acid, 100–500 ng for lupeol, and 500–2500 ng per spot for stigm...

Chemical constituents from fresh tubers of Dioscorea bulbifera

Eighteen compounds were isolated from the 95% ethanol extract of fresh tubers of Dioscorea bulbifera by column chromatography over silica gel,Sephadex LH-20, and ODS. Their structures were elucidated by spectroscopic data analysis as 6-hydroxy-2,10,10-trimethoxy-anthracen-9-one(1), diosgenin (2), stigmasterol(3), 3, 7-dimethoxy-5, 3', 4'-trihydroxyflavone(4), 2, 7-dihydroxy-3, 4-dimethoxyphenanthrene(5), 3, 7-dihydroxy-2, 4-dimethoxy phenanthrene(6), 2, 7-dihydroxy-4-methoxyphenanthrene (7), 2, 7-dihydroxy-3, 4-dimethoxy-9, 10-dihydroxy phenanthrene(8), azelaic acid (9), 8-epidiosbulbin E acetate (10), 1, 7-bis-(4-hydroxyphenyl)-4E, 6E-heptadien-3-one(11), diosbulbin B(12), pentacosanoic acid 2', 3'-dihydroxypropyl ester(13), 2, 7-dihydroxy-4-methoxy-9, 10-dihydroxy-phenanthrene (14), 1, 7-bis-(4-hydroxyphenyl)-1E, 4E, 6E-heptatrien-3-one (15), 6-ethoxy-1H-pyrimidine-2, 4-dione (16), 3, 5, 4'-trihydroxy-bibenzyl (17), and diosbulbin F (18). Compound 1 is a new compound, and compounds 7, 9, 13, and 16 were isolated from this plant for the first time.

Anti-diabetic activity of stigmasterol from soybean oil by targeting the GLUT4 glucose transporter

ABSTRACTThe present study investigated the anti-diabetic activity and potential mechanism of stigmasterol (SMR), which is a kind of phytosterols derived from the edible soybean oil in vitro and in vivo. SMR displayed a mild GLUT4 translocation activity by 1.44-fold in L6 cells. L6 cells were treated with different concentration of SMR, showing significant effects on the enhancing glucose uptake. SMR administrated orally to the KK-Ay mice significantly alleviated their insulin resistance and oral glucose tolerance with reducing fasting blood-glucose levels and blood lipid indexes such as triglyceride and cholesterol. Moreover, the GLUT4 expression in L6 cells, skeletal muscle and white adipose tissue had been also enhanced. In this paper we conclude that, stigmasterol seems to have potential beneficial effects on the treatment of type 2 diabetes mellitus with the probable mechanism of targeting GLUT4 glucose transporter included increasing GLUT4 translocation and expression.

Antimicrobial Analysis and Structural Elucidation Of Active Compounds Of Dialium Indum Leaves Extract (Valvet Tarmarind)

Antimicrobial analysis and structural elucidation were carried out on the purified leaves of Dialium indum. The Harbone method was used for the extraction. The extracts were separated using a combination of column chromatography and thin layer chromatography, which gave rise to the isolation of two fractions, these fractions were further purified using recrystallization. The melting point of each pure fraction was determined. The purified extracts were subjected to structural elucidation using various spectroscopic techniques which includes; FTIR, UV, H1 NMR, C13 NMR, DEPT 1350, COSY, TOCSY, HMBC and HSQC . The spectral analysis suggested the presence of Stigmasterol, and Lauric acid. The antimicrobial analysis (anti fungal and anti bacterial analysis) using the punched agar diffusion method was carried out on the isolated fractions comparatively with a standard drug Funbact-A cream (a broad spectrum antibiotic). A total of thirteen test organisms were used for this analysis amongst which were ten bacteria test organism and three fungi test organisms. The results from the average diameter zones of inhibition, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and fungicidal concentrations (MFC) showed that all the fractions were all active on the entire test organism with zones of inhibition ranging from 14mm-36mm comparatively. None of these fractions showed similar antimicrobial effect as the standard drug Funbact-A cream but individually could serve as anti microbial to diseases caused by these test organisms from their MIC, MBC and MFC.

Half-life of plasma phytosterols in very low birth weight preterm infants on routine parenteral nutrition with vegetable oil-based lipid emulsions

Phytosterols in vegetable oil (VO)-based lipid emulsions (LE) likely contribute to parenteral nutrition-associated cholestasis (PNAC) in preterm infants. No characterization of plasma phytosterol half-lives has been done in very low birth weight (VLBW) preterm infants receiving parenteral nutrition (PN) with LE. In a prospective cohort study, 45 VLBW preterm infants who received PN underwent serial blood sample measurements of sitosterol (SITO), campesterol (CAMP), and stigmasterol (STIGM). Plasma phytosterol half-lives were calculated from the phytosterol concentrations-decay curves by using a single-compartment model. After the stop of the intravenous LE, study infants had significantly lower plasma total CAMP, STIGM and SITO concentrations. The decay of plasma phytosterol concentrations was monoexponential. Half-life of plasma total CAMP, STIGM and SITO was 13.5±6.9, 10.3±4.5 and 10.3±4.0 days, respectively. Plasma phytosterol half-lives did not correlate with gestational age, birth weight, cumulative phytosterol intakes and plasma conjugated bilirubin. VLBW preterm infants on PN with LE had rather long plasma phytosterol half-lives similar to hypercholesterolemic adults and phytosterolemic homozygotes patients. We speculate that the accumulation of phytosterols could contribute to their vulnerability to PNAC. The Ethics Committee of Marche-Italy (DG/469); www.clinicaltrials.gov (identification number NCT02758834).

Phytochemical composition and antioxidant activity of extracts of some medicinal plants in Tunisia

The aim of this study was to investigate the chemical composition of the best known plants biomarkers from the locality of Gafsa (Tunisia). Majority of investigated plants had high levels of phenolics and exhibited noteworthy antioxidant activity. The plants showing the highest antioxidant activities were P. angustifolia, T. garganica, G. alypum and L. multifida. P. angustifolia demonstrated by far the most potent antioxidant activity, as well as total phenolic and total flavonoids content. In flavonoid structure of the aerial parts of the related species, different amounts of rutin, myricetin, morin, quercetin and kaempferol together with ergosterol, stigma sterol and b-sitosterol as phytosterols, vitamin D, vitamin K, a-tocopherol and retinol as vitamins, and arabinose, fructose, glucose, sucrose and maltose as sugars were found. The mineral analysis was related to the existence of Ca, Na, K, Fe, Mn, Zn and Mg with different amounts in all of the nine tested species.

Anthropogenic pressure in a Portuguese river: Endocrine-disrupting compounds, trace elements and nutrients

ABSTRACTNatural organic compounds such as phytoestrogens and phytosterols found in various plants, as well as mycotoxins produced by fungi, can be found in aquatic environments. The aim of this study was to investigate the occurrence of three different classes of natural estrogenic compounds, i.e., phytoestrogens, phytosterols and mycotoxins, in estuarine water samples from the Ave River estuary. For that, water samples were collected at five sampling points distributed along the estuary at low tide, during 1 year, processed by solid-phase extraction (SPE) and analyzed by gas chromatography coupled to mass spectrometry (GC–MS). To correlate the presence of phytoestrogens and phytosterols in the estuarine water, local flora was collected on riverside. Trace elements content and physicochemical parameters such as nutrients and dissolved oxygen were also determined seasonally at each sampling point, to give insights for the evaluation of water quality and anthropogenic pressure. Both phytoestrogens and phytosterols showed a seasonal variation, with the highest values observed in spring and summer and the lowest in winter. Daidzein (DAID) was found up to 404.0 ng L−1 in spring and coumestrol (COUM) was found up to 165.0 ng L−1 in summer. The mycotoxin deoxynivalenol (DON) was ubiquitously determined with values ranging from 59.5 to 642.4 ng L−1. Nutrients and metals distribution and concentration varied among sampling stations and seasons. This study revealed for the first time the presence of mycotoxins, various classes of phytoestrogens and stigmasterol (STG) in estuarine water from the Ave River (Portugal), and the evaluation of the water quality confirmed that this estuary is still highly impacted by anthropogenic activities.

English

Two major compounds, stigmasterol (ST) and sitosterol-3-O-β-D-glucopyranoside (SG) were isolated from Pseuderanthemum palatiferum leaf extract which is used traditionally as an antidiabetic. ST and SG at doses of 0.25 and 0.50 mg/kg were fed to diabetic rats for 21 days, and the fasting blood glucose (FBG) level and biochemical data on day 0, 4, 7, 10, 14, 17 and 21 were determined and compared with the anti-diabetic drug, glibenclamine. FBG levels at all doses of ST and SG were significantly decreased (p<0.05) with a concomitant increase in serum insulin. SG at the dose of 0.50 mg/kg showed the highest hypoglycemic effect. ST and SG also improved the following biochemical data and hematology parameters such as total cholesterol, triglycerides, high density lipoprotein (HDL), low density lipoprotein (LDL), blood urea nitrogen, creatinine, red blood cells, platelet and white blood cells. Key words: Pseuderanthemum palatiferum, acanthaceae, stigmasterol, sitosterol-3-O-β-D-glucopyranoside;, hypoglycemic activity.

Simultaneous Estimation of Stigmasterol and Withaferin A in Union Total Herbal Formulation Using Validated HPTLC Method

Triterpenoids and phytosteroids from stem bark of Crataeva nurvala buch ham.Shumaia Parvin, Md. Abdul Kader, Md. Abdul Muhit, Md. Ekramul Haque, Md. Ashik Mosaddik, Mir Imam Ibne Wahed

Determination of sterols using liquid chromatography with off-line surface-assisted laser desorption/ionization mass spectrometry

A new method, reversed phase liquid chromatography with off-line surface-assisted laser desorption/ionization mass spectrometry (RPLC–SALDI MS) for the determination of brassicasterol (BR), cholesterol (CH), stigmasterol (ST), campesterol (CA) and β-sitosterol (SI) in oil samples has been developed. The sample preparation consisted of alkaline saponification followed by extraction of the unsaponificable fraction with diethyl ether. The recovery of the sterols ranged from 91 to 95% with RSD less than 4%. Separation of the five major sterols on a C18 column using methanol-water gradient was achieved in about 10min. An on-line UV detector was employed for the initial sterol detection prior to effluent deposition using a laboratory-built spotter with 1:73 splitter. Off-line SALDI MS was then applied for mass determination/identification and quantification of the separated sterols. Ionization of the nonpolar analytes was achieved by silver ion cationization with silver nanoparticles used as the SALDI matrix providing limits of detection 12, 6 and 11fmol for CH, ST and SI, respectively. Because of the incorporated splitter, the effective limits of detection of the RPLC–SALDI MS analysis were 4, 3 and 4pmol (or 0.08, 0.06 and 0.08μg/mL) for CH, ST and SI, respectively. For quantification, 6-ketocholestanol (KE) was used as the internal standard. The method has been applied for the identification and quantification of sterols in olive, linseed and sunflower oil samples. The described off-line coupling of RPLC to SALDI MS represents an alternative to GC–MS for analysis of nonpolar compounds.

Phytochemistry and pharmacology of Tecomella undulata

Tecomella undulata (Bignoniaceae) is a monotypic genus and one of the most important deciduous, ornamental shrub or small tree of the acrid zone of India. Locally known as Rohida, Roheda in Hindi, Rakhtroda in Marathi, Dadimacchada, Chalachhada, Dadimapuspaka in Sanskrit mostly found in the Thar desert regions of India and Pakistan. The plant holds tremendous potential of medicinal value and is used in traditional and folklore system of medicines. It has been used traditionally in various ailments like syphilis, swelling, leucorrhoea and leucoderma, enlargement of spleen, obesity, tumours, blood disorders, flatulence and abdominal pain. Tecomella undulata has gained prominence due to presence of some prominent secondary metabolites of great therapeutic potential like stigmasterol, β‑sitosterol, α‑lapachone, tectol isolated from heartwood, bark and leaf. The present review presents the traditional information and recent scientific update on this plant with therapeutic potential. Key words: Hepatoprotective, pharamacology, phytochemistry, Tecomella undulata

Determination of sterol and fatty alcohols in unsaponifiable matter of Roystonea regia fruits oil

Roystonea regia (Kunth) F. Cook fruit is a good source of oil with possible application in the nutritional and pharmaceutical industries. The chemical constituent of the unsaponifiable matter of this oil, however, has not been studied yet. The aim of this work was to assess the content of sterols and alcohols in this fraction using GC-MS. Samples were submitted to basic hydrolysis with KOH/EtOH solution and then the unsaponifiable fractions extracted with n-hexane, and then alcohols and sterols were analyzed as trimethylsilylether derivatives using cholestane as the internal standard. The average content of the unsaponifiable matter of R. regia was 1.3%. The major fraction (66%), was composed of sterols, like s-sitosterol, stigmasterol, campesterol, 24-methylencycloartanol, Δ5-avenasterol, cycloartanol and cycloartenol, while the fatty alcohol fraction (33.9%) contained C18OH-C34OH alcohols.

Chemical constituents from the aerial parts of Salsola inermis

Background and objective The hydroalcoholic extract from the aerial parts of Salsola inermis exhibited antioxidant, anti-inflammatory, and antinociceptive effects. The present study deals with the isolation and identification of the chemical constituents of this hydroalcoholic extract. Materials and methods The aerial parts of S. inermis (Forsskal) were collected from wild plants growing near the El-Alamein area in October 2005. Air-dried and powdered aerial parts of S . inermis were extracted with 70% alcohol in H 2 O. The extract was partitioned successively with CHCl 3 , EtOAc, and n -BuOH. The structures of the isolated compounds were determined by chemical and spectroscopic analyses. Results and conclusion Phytochemical investigation of the alcoholic extract from the aerial parts of S. inermis revealed 12 compounds, identified as long chain hydroxyl fatty acid 9,12,13-trihydroxydecosan–10,15,19-trienoic acid; trans - N -feruloyl tyramine-4ooo- O -β-D-glucopyranoside; umbelliferone; scopoletin; 3-methyl kaempferol; olean-12-en-3,28-diol; olean-12-en-28-oic acid; stigmasterol-3-β- O -D-glucopyranoside; 3- O -[β-D-glucopyranosyl]oleanolic acid; kaempferol 3-O-β-glucopyranoside; and isorhamnetin 3- O -β-glucopyranoside, in addition to β-sitosterol, stigmasterol, and stigmastanol. Some of these compounds have hydroxyl groups, which help in scavenging free radicals and inhibit COX and various mediators involved in the pathogenesis of pain relief.

Distribution of sterols and the sources of pollution in surface sediments of Ulungur lake, Xinjiang

Domestic sewage discharged into lakes brings great pressure to the ecological environment. This study selected sediment from an inland lake as a research object to evaluate pollution of the environment. Eight sterols were used to evaluate the content of pollutants, while the ratios of sterols were used as the index to analyze the sources of pollution. The correlations were analyzed between sterols and total organic carbon (TOC), salinity and particle size. The distribution and composition of sterol compounds were determined in 12 surface sediment samples collected from Ulungur lake. The total concentrations of detected sterols in the sediments ranged from 1.3 to 36.3 μg/g.dw. The most abundant sterol detected was β-sitosterol (STI) with average concentrations of 2.6 μg/g.dw, followed by cholesterol (CHOE), stigmasterol (STIG) and stigmastanol (STAN). The concentration of coprostanol (COP) was between 0.03 and 1.66 μg/g.dw. Through correlation analysis, it was found that there was a significant correlation between fecal sterols and plant sterols. So the plant sterols shall not be neglected in evaluating the sources of pollution for their impact to identify the fecal sources. The study suggests that the composition and distribution of sterols in surface sediment provide useful information for environmental contamination monitoring and assessment in the inland lake.