Lipophilic Constituents Research Articles

Antioxidant property of Eucalyptus globulus Labill. Extracts and inhibitory activities on carbohydrate metabolizing enzymes related to type-2 diabetes

This research evaluated the antioxidant property and carbohydrate metabolizing enzyme inhibitory activity of Eucalyptus globulus Labill leaf extracts. Ethanol extracts of the prior hexane-defatted (DEE) and the non-defatted (NEE) leaf samples of the plant were prepared and screened for constituent phytochemicals by HPLC-DAD. The extracts were tested for antioxidant activity, α-amylase and α-glucosidase inhibitory activities and molecular docking with selected protein targets were carried out. The two extracts showed notable antioxidant property. Lipid peroxidation inhibitory activities of DEE, NEE and ascorbic acid were 28.4 ± 1.1 μg/ml, 33.3 ± 1.1 μg/ml, and 34.5 ± 1.2 μg/ml, respectively, in terms of IC 50 . The extracts also demonstrated appreciable inhibition of the carbohydrate metabolizing enzymes when compared to acarbose. IC 50 values for α-amylase inhibition were 23.6 ± 1.2 μg/ml, 14.8 ± 1.2 μg/ml and 5.2 ± 1.3 μg/ml for DEE, NEE and acarbose, respectively ( p < 0.05), while IC 50 values for α-glucosidase inhibition were 30.0 ± 1.1 μg/ml, 6.5 ± 1.2 μg/ml and 29.8 ± 1.8 μg/ml for DEE, NEE and acarbose, respectively ( p < 0.05). HPLC analysis of the extracts indicated that the defatting process eliminated pinitol and robinetinidol, and reduced the concentration the remaining phytoconstituents in the defatted extract. Catechin, ergosterol, pinitol, quercetin and robinetinidol identified by HPLC from the defatted and non-defatted extract displayed good interaction with the carbohydrate metabolizing enzymes with ergosterol presenting the best interaction. These results show that both extracts could be explored in alternative therapeutic approaches for type-2 diabetes. The non-defatted extract appeared to demonstrate superior activities suggesting a positive contribution from the lipophilic constituents of the plant. • Eucalyptus globulus leaf extracts show notable antioxidant activity. • The extracts also displayed good in vitro antidiabetic activities. • Lipophilic constituents enhanced the observed activities of E. globulus leaves. • Lipophilic constituents also contribute to the in vitro antidiabetic activity. • Molecular docking analysis reveal ergosterol as a principal bioactive compound.

Main Applications of Cyclodextrins in the Food Industry as the Compounds of Choice to Form Host-Guest Complexes.

Cyclodextrins (CDs) are cyclic oligomers broadly used in food manufacturing as food additives for different purposes, e.g., to improve sensorial qualities, shelf life, and sequestration of components. In this review, the latest advancements of their applications along with the characteristics of the uses of the different CDs (α, β, γ and their derivatives) were reviewed. Their beneficial effects can be achieved by mixing small amounts of CDs with the target material to be stabilized. Essentially, they have the capacity to form stable inclusion complexes with sensitive lipophilic nutrients and constituents of flavor and taste. Their toxicity has been also studied, showing that CDs are innocuous in oral administration. A review of the current legislation was also carried out, showing a general trend towards a wider acceptance of CDs as food additives. Suitable and cost-effective procedures for the manufacture of CDs have progressed, and nowadays it is possible to obtain realistic prices and used them in foods. Therefore, CDs have a promising future due to consumer demand for healthy and functional products.

Tanshinone IIA: A Review of its Anticancer Effects.

Tanshinone IIA (Tan IIA) is a pharmacologically lipophilic active constituent isolated from the roots and rhizomes of the Chinese medicinal herb Salvia miltiorrhiza Bunge (Danshen). Tan IIA is currently used in China and other neighboring countries to treat patients with cardiovascular system, diabetes, apoplexy, arthritis, sepsis, and other diseases. Recently, it was reported that tan IIA could have a wide range of antitumor effects on several human tumor cell lines, but the research of the mechanism of tan IIA is relatively scattered in cancer. This review aimed to summarize the recent advances in the anticancer effects of tan IIA and to provide a novel perspective on clinical use of tan IIA.

Comprehensive metabolic profiling of modified gegen qinlian decoction by ultra-high-performance liquid chromatography-diode array detection-Q-exactive-orbitrap-electrospray ionization-mass spectrometry/mass spectrometry and application of high-performance thin-layer chromatography for its fingerprint analysis

Objective: Gegen Qinlian decoction (GQD) is a classical traditional Chinese medicine formulation which has been used for almost 2000 years. At Guang'anmen Hospital, Beijing, a modified GQD version (mGQD) with seven instead of four herbal ingredients has been applied to treat Type 2 diabetes. Quality control is a crucial prerequisite for the therapeutic application of herbal medicines. For the identification of products derived from classical GQD, the Chinese Pharmacopeia requires the analysis of only three marker compounds. Because mGQD is a more complex mixture containing seven herbs and hundreds of constituents, the pharmacopoeia method for GQD is inadequate. Materials and Methods: A more comprehensive characterization of the formula's constituents has been developed using ultra-high-performance liquid chromatography-diode array detection (UHPLC-DAD)-Q-Exactive-mass spectrometry (MS) in electrospray ionization positive and negative mode. Moreover, a new method for the fingerprint analysis of mGQD via high-performance thin-layer chromatography (HPTLC) has been established. Results: Altogether, 91 compounds have been assigned to their originating plants and 84 substances were identified either by comparison with authentic references or with data from the literature. The HPTLC method is based on the application of two different mobile phases and is able to detect both lipophilic and hydrophilic constituents of mGQD. Conclusions: The modified GQD was extensively characterized by UHPLC combined with DAD and Q-Exactive Orbitrap high-resolution MS detection, leading to the assignment and identification of compounds present in the decoction. In addition, a new method for the fingerprint analysis of the mGQD using HPTLC was established, which allows fast and simple identification of the herbal ingredients in the mixture.

Gastric lipase can significantly increase lipolysis and carotenoid bioaccessibility from plant food matrices in the harmonized INFOGEST static in vitro digestion model.

Gastrointestinal digestion of carotenoids has received much attention, as these lipophilic compounds have been related to several health benefits. Most commonly, static digestion models such as the consensus INFOGEST model are employed to study their bioaccessibility from test matrices. However, an aspect that has been much neglected is the use of gastric lipase. Its inclusion to gastro-intestinal (GI) digestion is expected to foster emulsification of lipophilic constituents prior to their incorporation into mixed micelles. In this study, we compared the effect of various lipases from R. niveus, R. oryzae, and rabbit gastric extracts (RGE), at different concentrations (0, 30, and 60 U mL-1), on carotenoid bioaccessibility from several food matrices (tomato juice, spinach, and carrot juice). We also investigated whether co-digestion of pure proteins (whey and soy protein isolates) at 0, 25, and 50% of the equivalent recommended dietary allowance, would interact with carotenoid bioaccessibility in presence or absence of RGE. Lipolysis was also studied. Considering all matrices combined, lipases significantly improved the bioaccessibility of carotenoids (p < 0.001). Compared to other lipases, RGE consistently increased carotenoid bioaccessibility in all tested matrices, by up to 182% (p < 0.001), this effect was partly maintained in the presence of co-digested proteins. Unexpectedly, all 3 lipases improved gastric lipolysis in all matrices, by an average of 10-fold (p < 0.001). In conclusion, only RGE contributed significantly to improving both lipolysis extent and carotenoid bioaccessibility in all tested matrices, while the presence of proteins mitigated the positive effect of lipases on carotenoid bioaccessibility.

The potentiation of beta-lactam and anti-bacterial activities of lipophilic constituents from Mesua ferrae leaves against methicillin-resistant Staphylococcus aureus

Abstract Objectives Mesua ferrae, from the family of Calophyllaceae, is traditionally used for the treatment of piles, fever and renal disorders. The present study was aimed to examine the antibacterial compounds from the leaves of M. ferrae and their β-lactam antibiotic potentiate activities against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA). Methods Stigmasterol (1) and β-caryophyllene oxide (2) were isolated from the n-hexane fraction of the leaves of M. ferrae using a bioassay-guided fractionation approach. Results The isolated compounds displayed anti-Staphylococcus and anti-MRSA activities. It is worth to note that both compounds demonstrated synergism with β-lactam antibiotics against S. aureus and MRSA. Gas chromatography–mass spectrometry (GC–MS) analysis indicated the n-hexane fraction was dominated by triterpenes and sesquiterpenes, suggesting the total antibacterial activity exhibited by the fraction. Conclusion Based on the findings, it could conclude that M. ferrae is a promising natural source for the discovery of new anti-MRSA lead compounds.

Bioactive Components in Methyl tert-Butyl Ether Extract of Sea Buckthorn (Hippophae rhamnoides L.) Green Waste

Lipophilic constituents of sea buckthorn leafy shoots, which is a large-scale waste product of sea buckthorn oil production and rejuvenating pruning of cultivated sea buckthorn plants, is investigated. Methyl tert-butyl ether (MTBE) is used as an extraction solvent. It features all the advantages of diethyl ether while lacking its disadvantages: it is not prone to peroxide formation and is not a significant air pollutant due to its higher boiling point. The chemical composition of MTBE extract of sea buckthorn leafy shoots is studied by gas chromatography mass spectrometry and high-performance liquid chromatography. Seventy-four neutral and 38 acidic components, including polyprenols, dolichols, triterpene alcohols and acids, and sterols are identified. In comparison to hexane, MTBE, as an extraction solvent, provides higher yields of bioactive triterpene acids and diols, including uvaol, eritrodiol, and betulin. The presence of these compounds in the prepared extract may enhance its beneficial properties.

Wax-based delivery systems: Preparation, characterization, and food applications.

The development of lipid-based delivery systems has attracted much attention over the last years and a wide variety of strategies and formulations are currently available to encapsulate, protect, and target delivery of bioactive and functional lipophilic constituents within the food and pharmaceutical industries. Waxes are crystalline lipid material, consisting of a complex mixture of long-chain fatty acids and fatty alcohols, hydrocarbons, aldehydes, and ketones and show great promises as constituents of carrier systems. Most of waxes are classified under food-grade category and show high availability at a low cost. This review article has provided a comprehensive summary of research on major carriers containing wax as one of the main constituents, including solid lipid nanoparticles, nanostructured lipid carriers, oleogels, and Pickering emulsions, with a focus on their food applications. The physical and chemical nature of natural waxes are described in the first while the second part deals with the structure, formulation, main methods of preparation, characterization, and finally utilization of each type of wax-based delivery system for specific food applications.

Characterization of nutritionally important lipophilic constituents from brown kelp Ecklonia radiata (C. Ag.) J. Agardh

This research study presents information for the first time on the nutritionally relevant lipophilic compounds obtained from Ecklonia radiata, a poorly studied brown kelp. The major lipophilic compounds were analyzed utilizing liquid chromatography (LC)-tandem mass spectrometry (MS/MS) and gas chromatography (GC)-mass spectrometry (MS). The LC-MS/MS results revealed the presence of eight major lipophilic compounds, including sterols, carotenoids, vitamin E, and phylloquinone (vitamin K1). Quantitative analysis showed that fucosterol was the most predominant phytosterol in the fronds and stipes of E. radiata. The carotenoids (all-E)-fucoxanthin and (all-E)-β-carotene were present in higher yield. In terms of vitamin E, α-tocopherol was identified as the main tocol. The coenzyme, phylloquinone, important for protein synthesis, was also identified in E. radiata. GC–MS identified 13 fatty acids with palmitic (C16:0) and oleic acid (C18:1n9c) present in the highest quantities. To our knowledge, this is the first report on E. radiata, and the valuable data presented herein can be used as a baseline for developing novel nutraceuticals.

Identification of lipophilic components in Citri Reticulatae Pericarpium cultivars by supercritical CO2 fluid extraction with ultra-high-performance liquid chromatography-Q Exactive Orbitrap tandem mass spectrometry.

To systematically identify the lipophilic constituents of Citri Reticulatae Pericarpium from different cultivars, supercritical CO2 fluid extraction and ultra-high-performance liquid chromatography-Q Exactive Orbitrap tandem mass spectrometry were integrated for the component analysis of 18 batches of Citri Reticulatae Pericarpium from 12 cultivars for the first time. A total of 57 components from the supercritical CO2 fluid extracts were demonstrably or tentatively identified by the obtained parent peaks, fragment peaks, and retention times. In total, two flavonoids, six organic acids, nine coumarins, three aldehydes, seven esters, three terpenes, one limonoid, and five other compounds were detected for the first time; notably, coumarin components have not yet been reported in Citri Reticulatae Pericarpium. Furthermore, the extract constituents differed between cultivars. In particular, organic acids were more abundant in Citrus reticulata "Chachi" than in other cultivars, and pterostilbene was exclusively found in Citrus reticulata "Yichangju". The results showed that a greater variety of compounds in Citri Reticulatae Pericarpium could be extracted by supercritical CO2 fluid extraction and detected by ultra-high-performance liquid chromatography-Q Exactive Orbitrap tandem mass spectrometry. This study provides a more scientific basis for further analysis of the pharmacological activity and quality of Citri Reticulatae Pericarpium components from different cultivars.

Carotenoids, Fatty Acids, and Volatile Compounds in Apricot Cultivars from Romania-A Chemometric Approach.

Lipophilic constituents are important for the color and aroma of apricots, but also for their health benefits. In the present study, carotenoids, fatty acids, and volatiles were analyzed in 11 apricot cultivars, from which nine were obtained in Romania. High performance liquid chromatography coupled to a diode array detector with atmospheric pressure chemical ionization and mass spectrometry (HPLC-DAD-APCI-MS methodology applied on unsaponified carotenoid extracts allowed the identification and quantification of 19 compounds. The predominant carotenoids in all cultivars were all-trans-β-carotene and its cis isomers. Lutein was present exclusively in non-esterified form, while β-cryptoxanthin was predominantly esterified, mainly with oleic, palmitic, lauric, and stearic acid. Moreover, β-cryptoxanthin linoleate, linolenate, and stearate were detected for the first time in Harogem cultivar. Variation in carotenoid content and composition was observed, with the highest carotenoid content being recorded in Tudor, Harogem, and Mamaia cultivars. The predominant fatty acids determined by gas chromatography–mass spectrometry (GC-MS) were linoleic (up to 47%), palmitic (up to 32.7%), and linolenic (up to 17.16%), with small variations among cultivars. In-tube extraction technique (ITEX)/GC-MS was applied for profiling the volatiles in apricot fruits and 120 compounds were identified, with terpenoids and esters as the most abundant classes. Principal component analysis (PCA) revealed that the carotenoids and the fatty acids profile can be used for variety authentication and discrimination in apricots.

Isolation and determination of lipophilic mycochemicals from a New Zealand edible native mushroom Hericium novae-zealandiae

Hericium novae-zealandiae is a native mushroom consumed by indigenous Māori people in New Zealand. The lipophilic mycochemicals of the mushroom were isolated using a normal column chromatography combined with a preparative HPLC. Structural characterisation based on spectroscopic methods, namely UV, MS, NMR and single crystal XRD have identified three lipophilic compounds as hericene B (a compound unique to Hericium), ergosterol and ergosterol peroxide. Following this, an HPLC-DAD method was developed and validated to quantify the hericene B and ergosterol. The method showed excellent selectivity, linearity, precision, accuracy and robustness. The content of hericene B was determined as 28.53 mg/g by dry weight of H. novae-zealandiae (approximately 3%). This discovery indicates the potential utilisation of H. novae-zealandiae as a natural source of hericene B. Current research revealed for the first time, the lipophilic constituents of H. novae-zealandiae and the method development for quantification of hericene B in the above species.

LIPOPHILIC CONSTITUENTS OF PEDICULARIS STRIATA PALLAS И PEDICULARIS FLAVA PALLAS

Some species of Pedicularis have been widely applied in traditional Chinese medicine. Some plants of Pedicularis genus have a great potential for the discovery of new drugs because most of them have not been studied in detail. Extracts from the plants of genus Pedicularis have been found to possess antitumor, hepatoprotective, anti-oxidative, antihaemolysis, antibacterial activity, fatigue relief of skeletal muscles, nootropic effect and other activitiesin vivo and in vitro. A wide range of chemical components including iridoid glycosides, phenylpropanoid glycosides, lignans glycosides, flavonoids, alkaloids and other polar compounds have been isolated and identified from the genus Pedicularis. Information about lipophilic constituents is too poor. The results of our investigations of aliphatic and triterpenoic substances by GC-MS are identification of 25 neutral and 18 acid compounds including sterols, triterpenoids and unusual branched hydrocarbons. Main components of neutral fraction are β-sitosterol and phytol. Triterpenol fraction is found to content cycloartenol, 24-methylene-cycloartanol, obtusifoliol, stigma-7-en-3-ol and citrostadienol. Cycloartenol prevails in both types of row material. The main components of acids are palmitic, linoleic and linolnic acids. Pedicularis striata contains significant amount of melissic and montanic acids. All compounds were found for the first time in these row materials. Identified compounds could be potential chemotaxonomic markers for the Pedicularis species.

Fingerprinting of cerrado species based on cork lipophilic constituents

Cerrado is a savanna ecosystem of central and southeastern Brazil. Many woody species of cerrado have thick layers of cork. The present work aimed to characterize, by GC/MS analysis, the constituents of n-hexane extracts from the cork of common species from cerrado. Cork samples from 31 individuals, corresponding to 14 species and six families, were analyzed. Similarities and differences were noticed between cork and cuticular waxes regarding profiles of lipophilic constituents. The distribution of cork constituents was analyzed using the UPGMA clustering method and DICE coefficient. All clusters in the dendrogram obtained comprise individuals from a same species, suggesting that the distribution of lipophilic cork constituents is useful for species characterization and possibly also for species identification, resembling results commonly obtained with molecular markers. Seven samples of Bignoniaceae, corresponding to two genera and seven species, emerged in a common cluster, in an arrangement in accordance with the recent segregation of Tabebuia species to a new genus Handroanthus. The markers analyzed were not efficient regarding characterization of other families.

Lipophilic extractives of the wood and bark from Eucalyptus pellita F. Muell grown in Merauke, Indonesia

Lipophilic extractives in the dichloromethane extract from Eucalyptus pellita wood and bark were identified by GC-MS. Of the various morphological parts, the bark samples contain more extractives than wood samples. The largest extractive content was measured in the bark of the top part. Four groups (i.e., free fatty acids, sterols-steroids, triterpenoids, and other compounds) were detected in all parts. Palmitic, oleic, and linoleic acids as well as β-sitosterol dominated among of the lipophilic extractives. Long chain fatty acids were the major lipophilic groups in heartwood. Sterols-steroids were the major components in the bottom and top part of the bark. Conversely, triterpenoids and short chain-fatty acids were the minor components in bottom and top part of the bark. Very long-chain fatty acids (i.e., docosanoic, lignoceric, and hexacosanoic) were also observed. The concentration of certain lipophilic constituents, e.g., stigmast-4-en-3-one, squalene, glycerol, oleic, linoleic, hexacosanoic, and β-sitosterol, was affected by position within the tree in axial and radial directions. The higher concentration of long chain fatty acids in the heartwood as well as sterols-steroids in the bark seems to be clearly disadvantage as far as pitch formation is concerned.

Lipophilic compounds, but not fucoxanthin, mediate the genotoxic effect of photoautotrophic grown Phaeodactylum tricornutum in Caco-2 and HT-29 cells

Abstract Phaeodactylum tricornutum (P. triconutum) gained high interest for use in human nutrition. We aimed to investigate the effect of photoautotrophically cultivated Phaeodactylum tricornutum on genotoxicity, cytotoxicity, formation of reactive oxygen species and apoptosis in Caco-2 and HT-29 cells. Furthermore, the effect of processing (sonication, ball-milling) on these parameters was assessed. P. triconutum dose-dependently induced DNA strand breaks (2 µg dry matter/mL – 200 µg dry matter/mL) in both cell lines. These breaks can mostly be repaired after a recovery time of 2 h. Using processing methods did not affect the genotoxicity. We found that a lipophilic fraction, but not fucoxanthin, might be responsible for the DNA damage. Neither the P. triconutum preparations nor fucoxanthin affected cell proliferation or revealed an apoptotic or necrotic activity. In conclusion, P. triconutum showed a genotoxic potential in intestinal cells, mainly mediated by lipophilic constituents. Sonication and ball-milling might be considered as safe methods for processing of P. triconutum.

Modeling and optimization of supercritical carbon dioxide extraction for isolation of valuable lipophilic constituents from elderberry (Sambucus nigra L.) pomace

Supercritical carbon dioxide extraction (SFE-CO2) was employed and its parameters (pressure, temperature, time) were optimized in order to recover valuable non-polar constituents from elderberry juice processing by-products (pomace). Several other commonly applied hexane-utilizing fractionation techniques, namely pressurized liquid (PLE), ultrasound-assisted (UAE), Soxhlet and conventional solid-liquid (SLE, maceration) extractions, were used for comparison purposes. Under optimal SFE-CO2 conditions (53 °C, 35 MPa, 45 min), 14.05 g of the lipophilic fraction was recovered from 100 g of pomace, containing health beneficial polyunsaturated linoleic (42.0%) and α-linolenic (34.1%) fatty acids. In terms of extraction yields and time, the efficiency of SFE-CO2 was generally higher as compared to conventional Soxhlet and SLE, but lower than PLE and UAE. The cyanogenic glycoside sambunigrin content in elderberry pomace, all lipophilic extracts and extraction residues was very low (6.7–76.6 ng/100 g pomace) as compared to the EFSA’s acute reference dose for HCN (20 μg/kg BW). Generally, a small portion of antioxidants was recovered from elderberry pomace after lipophilic fractionation either with supercritical CO2 or hexane (TPC: 1.5–4.7 mg GAE/g, TEAC: 0.3–11.6 mg TE/g) and defatted elderberry pomace residues retained a considerable amount (>60%) of these bioactives.

Nanoparticulated Honokiol Mitigates Cisplatin-Induced Chronic Kidney Injury by Maintaining Mitochondria Antioxidant Capacity and Reducing Caspase 3-Associated Cellular Apoptosis.

Cisplatin is a potent anti-cancer drug, however, its accompanied organ-toxicity hampers its clinical applications. Cisplatin-associated kidney injury is known to result from its accumulation in the renal tubule with excessive generation of reactive oxygen species. In this study, we encapsulated honokiol, a natural lipophilic polyphenol constituent extracted from Magnolia officinalis into nano-sized liposomes (nanosome honokiol) and examined the in vivo countering effects on cisplatin-induced renal injury. We observed that 5 mg/kg body weight. nanosome honokiol was the lowest effective dosage to efficiently restore renal functions of cisplatin-treated animals. The improvement is likely due the maintenance of cellular localization of cytochrome c and thus preserves mitochondria integrity and their redox activity, which as a consequence, reduced cellular oxidative stress and caspase 3-associated apoptosis. These improvements at the cellular level are later reflected on the observed reduction of kidney inflammation and fibrosis. In agreement with our earlier in vitro study showing protective effects of honokiol on kidney cell lines, we demonstrated further in the current study, that nanosuspension-formulated honokiol provides protective effects against cisplatin-induced chronic kidney damages in vivo. Our findings not only benefit cisplatin-receiving patients with reduced renal side effects, but also provide potential alternative and synergic solutions to improve clinical safety and efficacy of cisplatin treatment on cancer patients.

Tanshinone IIA Exerts Anti-Inflammatory and Immune-Regulating Effects on Vulnerable Atherosclerotic Plaque Partially via the TLR4/MyD88/NF-κB Signal Pathway.

Background: Tanshinone IIA (Tan IIA), a lipophilic constituent from Salvia miltiorrhiza Bunge, has shown a promising cardioprotective effect including anti-atherosclerosis. This study aims at exploring Tan IIA’s anti-inflammatory and immune-regulating roles in stabilizing vulnerable atherosclerotic plaque in ApoE-deficient (ApoE−/−) mice. Methods: Male ApoE−/− mice (6 weeks) were fed with a high-fat diet for 13 weeks and then randomized to the model group (MOD) or Tan IIA groups [high dose: 90 mg/kg/day (HT), moderate dose: 30 mg/kg/day (MT), low dose: 10 mg/kg/day (LT)] or the atorvastatin group (5 mg/kg/day, ATO) for 13 weeks. Male C57BL/6 mice (6 weeks) were fed with ordinary rodent chow as control. The plaque stability was evaluated according to the morphology and composition of aortic atherosclerotic (AS) plaque in H&E staining and Movat staining sections by calculating the area of extracellular lipid, collagenous fiber, and foam cells to the plaque. The expression of the Toll-like receptor 4 (TLR4)/myeloid differentiation factor88 (MyD88)/nuclear factor-kappa B (NF-κB) signal pathway in aorta fractions was determined by immunohistochemistry. Serum levels of blood lipid were measured by turbidimetric inhibition immunoassay. The concentrations of tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCP-1) were detected by cytometric bead array. Results: Tan IIA stabilized aortic plaque with a striking reduction in the area of extracellular lipid (ATO: 13.15 ± 1.2%, HT: 12.2 ± 1.64%, MT: 13.93 ± 1.59%, MOD: 18.84 ± 1.46%, P < 0.05) or foam cells (ATO: 16.05 ± 1.26%, HT: 14.88 ± 1.79%, MT: 16.61 ± 1.47%, MOD: 22.08 ± 1.69%, P < 0.05) to the plaque, and an evident increase in content of collagenous fiber (ATO: 16.22 ± 1.91%, HT: 17.58 ± 1.33%, MT: 15.71 ± 2.26%, LT:14.92 ± 1.65%, MOD: 9.61 ± 0.7%, P < 0.05) to the plaque than that in the model group, concomitant with down-regulation of the protein expression of TLR4, MyD88, and NF-κB p65, and serum level of MCP-1 and TNF-α in a dose-dependent manner. There were no differences in serum TC, LDL, HDL, or TG levels between ApoE–/– mice and those treated with atorvastatin. Conclusions: These results suggest that Tan IIA could stabilize vulnerable AS plaque in ApoE−/− mice, and this anti-inflammatory and immune-regulating effect may be achieved via the TLR4/MyD88/NF-κB signaling pathway.

ALIPHATIC AND TRITERPENOIC PRODUCTS OF ETHER EXTRACTS SAPONIFICATION OF POPULUS NIGRA L.

Our investigation deals with extractives which content about 15% of whole biomass. The extracts obtained from poplar bark and foliage, and purified phytochemicals arising from these extracts, can find various applications, e.g., as biopharmaceuticals, food additives and nutraceuticals, biopesticides, and cosmetic products. Composition of neutral lipophylic compounds of poplar bark and foliage was studied as far as inflorescences of male and female plants and pollen. Tert-butyl methyl ether (TBME) was used as an extraction solvent for raw materials having all the advantages of diethyl ether, but devoid of its disadvantages. It does not form peroxides and does not create increased gas content due to the higher boiling point. TBME-extracts were investigated by gas chromatography–mass-spectrometry (GC-MS). We found more than 40 lipophilic neutral constituents from raw materials of poplar bark and foliage. 60 lipophilic neutral constituents from raw materials of florescences and pollen including alkanes, sterols, aliphatic and triterpenoic alcohols as far as 34 acidic compounds were identified by GC-MS. The content of inflorescences and pollen extracts differs from bark and foliage constituents.