Human Low-density Lipoprotein Oxidation Research Articles

Antioxidant effects of 14 Chinese traditional medicinal herbs against human low-density lipoprotein oxidation

The relationship between the antioxidant activities and inhibitory effect of 14 Chinese medicinal herbs against oxidized low-density lipoprotein (LDL) formation was evaluated. Prolongation of the lag phase of LDL oxidation depended on the concentration of the herbs. The concentration of each herb that was able to prolong the lag time by about two-fold was calculated and expressed as doubling-time concentration. The lower the doubling-time concentration, the stronger the inhibitory effect exhibited toward LDL oxidation. Among them, Chrysanthemi Flos (Chrysanthemum morifolium ramat; 甘菊花 gān jú huā), Crataegi Fructus (Crataegus pinnatifida Bge. var. major N.E.Br.; 山楂 shān zhā), and Roselle (Hibiscus sabdariffa Linn.; 洛神 luò shén) showed significant inhibitory effects. Correlation coefficients between doubling-time concentration and radical-scavenging activities were high; the total phenolic content was also high. In conclusion, phenolic compounds contributed not only to antioxidant activities, but also to the inhibitory effect against LDL oxidation. Chrysanthemi Flos, Crataegi Fructus, and H. sabdariffa, with lower doubling-time concentrations, could be potent phytochemical agents to reduce LDL oxidation and prevent the progression of atherosclerosis.

Phenolic Compounds of Cereals and Their Antioxidant Capacity

Phenolic compounds play an important role in health benefits because of their highly antioxidant capacity. In this review, total phenolic contents (TPCs), phenolic acid profile and antioxidant capacity of the extracted from wheat, corn, rice, barley, sorghum, rye, oat, and millet, which have been recently reported, are summarized. The review shows clearly that cereals contain a number of phytochemicals including phenolics, flavonoids, anthocyanins, etc. The phytochemicals of cereals significantly exhibit antioxidant activity as measured by trolox equivalent antioxidant capacity (TEAC), 2,2-diphenyl-1-picrylhydrazyl radical scavenging, reducing power, oxygen radical absorbance capacity (ORAC), inhibition of oxidation of human low-density lipoprotein (LDL) cholesterol and DNA, Rancimat, inhibition of photochemilumenescence (PCL), and iron(II) chelation activity. Thus, the consumption of whole grains is considered to have significantly health benefits in prevention from chronic diseases such as cardiovascular disease, diabetes, and cancer because of the contribution of phenolic compounds existed. In addition, the extracts from cereal brans are considered to be used as a source of natural antioxidants.

Antioxidant and Anti‐inflammatory Activities of Methanol Extracts of Tremella fuciformis and Its Major Phenolic Acids

Methanol extract subfractions of the edible white jelly mushroom (Tremella fuciformis), were assessed for the following antioxidant properties: ABTS(+) radical scavenging activity, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, and inhibitory activity of human low-density lipoprotein (LDL) oxidation. Among the subfractions tested, the chloroform subfraction exhibited the strongest antioxidant activity, with the highest total phenolic content (66.31 μg CAE/mg extract) and flavonoids content (5.12 μg QE/mg extract). The ABTS(+) radical scavenging activity of the chloroform subfraction was 7.89 μmol trolox/mg extract, which was the highest among all subfractions. This subfraction also showed the highest DPPH radical scavenging activity and inhibitory activity of LDL oxidation. In addition, the chloroform subfraction demonstrated anti-inflammatory activity through inhibition of nitric oxide production and inducible nitric oxide synthase expression in RAW 264.7 cells. Major phenolic acids from the mushroom extract were identified as 4-hydroxybenzoic acid (323 mg/kg dry weight of mushroom), gentisic acid (174 mg/kg dry weight of mushroom), and 4-coumaric acid (30 mg/kg dry weight of mushroom).

Inhibition of Human Low-Density Lipoprotein Oxidation In Vitro by Ginger Extracts

Oxidative modification of low-density lipoprotein (LDL) is thought to play a key role in atherosclerotic plaque formation. Currently, there is a renewed interest in ginger because of its antioxidants and cardioprotective properties. The effects of ethanol, methanol, ethyl acetate, and hexane solvent extracts of ginger and pure major ginger constituents on Cu(2+)-induced oxidation of human LDL in vitro were examined. The LDL oxidation inhibition by ethanol, methanol, ethyl acetate, and hexane extracts of ginger was 71%, 76%, 67%, and 67%, respectively, at their optimum extraction conditions. Inhibition of LDL oxidation by water extracts of ginger, which was prepared by ultrasonic-assisted extraction conditions of 52°C for 15 min, was about 43%. Phenolic bioactives of ginger-6-gingerols, 8-gingerols, 10-gingerols, and 6-shogaol-seem to be strong inhibitors of Cu(+2)-induced LDL oxidation. Overall, ginger extracts, including the water extract possess the antioxidant activities to inhibit human LDL oxidation in vitro.

Inhibitory effect on in vitro LDL oxidation and HMG Co-A reductase activity of the liquid-liquid partitioned fractions of Hericium erinaceus (Bull.) Persoon (lion's mane mushroom).

Oxidation of low-density lipoprotein (LDL) has been strongly suggested as the key factor in the pathogenesis of atherosclerosis. Mushrooms have been implicated in having preventive effects against chronic diseases due especially to their antioxidant properties. In this study, in vitro inhibitory effect of Hericium erinaceus on LDL oxidation and the activity of the cholesterol biosynthetic key enzyme, 3-hydroxy-3-methyl glutaryl coenzyme A (HMG Co-A) reductase, was evaluated using five liquid-liquid solvent fractions consisting of methanol : dichloromethane (M : DCM), hexane (HEX), dichloromethane (DCM), ethyl acetate (EA), and aqueous residue (AQ). The hexane fraction showed the highest inhibition of oxidation of human LDL as reflected by the increased lag time (100 mins) for the formation of conjugated diene (CD) at 1 µg/mL and decreased production (68.28%, IC50 0.73 mg/mL) of thiobarbituric acid reactive substances (TBARS) at 1 mg/mL. It also mostly inhibited (59.91%) the activity of the HMG Co-A reductase at 10 mg/mL. The GC-MS profiling of the hexane fraction identified the presence of myconutrients: inter alia, ergosterol and linoleic acid. Thus, hexane fraction of Hericium erinaceus was found to be the most potent in vitro inhibitor of both LDL oxidation and HMG Co-A reductase activity having therapeutic potential for the prevention of oxidative stress-mediated vascular diseases.

Phenolic Components of Dendrobium antennatum

The Chinese herbal medicine “shi-hu” (Dendrobii Caulis), prepared from the dried stem of Dendrobium species (Orchidaceae), has been used as a health tonic for fortification of the gastrointestinal system, the regulation of body fluid balance, as well as an antipyretic agent 1. Recent studies have further demonstrated its effect on smooth muscle cell proliferation, its use as an inhibitor of human low-density lipoprotein oxidation, in anti-angiogenesis, and in cytotoxicity [2, 3]. In the course of previous studies of chemical constituents of the Taiwanese Orchidaceae plants, including D. moniliforme [4, 5], D. clavatum var. aurantiacum [6], D. loddigesii [7], and D. sonia [8], phenanthraquinone, aromatics, long-chain fatty acids and esters, phytosterols, coumarin, bibenzyls, fluorenones, phenanthrenes, and sesquiterpenoids have been isolated from those plants. In our continuing research on the Dendrobium species to discovery the drug source of “shi-hu” cultivated in Taiwan [7, 8], we now report the phytochemical investigation of D. antennatum Lindl., an orchid cultivated in the garden.

Low-Density Lipoprotein (LDL) Antioxidants and Antiplatelet Compounds from Selected Malaysian Plants as Leads for the Development of Cardiovascular Protective Agents

Oxidative modification of low-density lipoprotein (LDL) plays a significant role in the onset and acceleration of atherosclerotic process and the development of cardiovascular diseases. The chronic antioxidant, hypolipidemic, antithrombotic and antiplatelet activities of many phytochemicals have important roles in prevention of lipoprotein oxidation and atherosclerotic lesion development. Phytochemicals of diverse chemical structures, including phenolic compounds, xanthones, prenylflavonoids, benzophenones, alkaloids and triterpenoids isolated from Malaysian plants were investigated for their ability to inhibit arachidonic acid (AA), collagen and adenosine diphosphate (ADP) induced platelet aggregation in human whole blood using an electrical impedance method. The compounds were also investigated for their ability to inhibit human LDL oxidation using thiobarbituric acid reactive substances (TBARS) assay. Among the compounds tested, 2-(3-methylbut-2-enyl)1,3,5,-trihydroxyxanthone, 2-(3-methylbut-2-enyl)-1,3,5,6-tetrahydroxyxanthone, rubraxanthone and 3,5,3′,5′-tetrahydroxy-4methoxybenzophenone isolated from Guttiferae species showed strong inhibition on platelet aggregation. Curcumin, cardamonin, pinocembrine, 5,6-dehydrokawain and 3-deacetylcrotepoxide were the strong antiplatelet compounds identified from Zingiberaceae species. Of the alkaloids tested, (-)-N-acetylnornuciferine, (-)-N-acetylanonaine and 1-(N-acetyl-Nmethylamino)ethyl-3,4,6-trimethoxy-7-hydroxyphenanthrene from Armodendron elegans showed strong inhibition on platelet aggregation caused by all three inducers. 3,5,3′,5′-Tetrahydroxy-4-methoxybenzophenone and 1, 7-dihydroxyxanthone showed strong inhibitory activity on LDL oxidation. Curcumin, demethoxycurcumin and bisdemethoxycurcumin exhibited stronger activity than the reference, probucol, as LDL antioxidants. Xanthorrhizol, zerumbone and ar-turmerone from the oils of Curcuma species showed relatively strong antioxidant activity. 3,5,3′,5′-Tetrahydroxy-4-methoxybenzophenone and 1, 7dihydroxyxanthone also showed strong inhibitory activity on LDL oxidation. Of the triterpenoids tested, garcihombronane B from Garcinia cantleyana showed moderate activity against LDL oxidation. The compounds are new classes of natural products that can be used as leads for development of new cardiovascular protective agents.

Inhibitory activity of Ecklonia stolonifera and its isolated phlorotannins against Cu2+-induced low-density lipoprotein oxidation

Since the oxidation of low-density lipoprotein (LDL) is one of the main causes of atherosclerosis, Cu2+-induced LDL oxidation and conjugated diene formation is currently being explored for the development of pharmaceutical drugs or functional foods for the treatment of atherosclerosis. The present work investigated the inhibitory effects on in vitro Cu2+-induced human LDL oxidation and conjugated diene formation of the methanol (MeOH) extract of the edible brown alga (Ecklonia stolonifera) and its different solvent-soluble fractions, as well as the phlorotannins isolated from them. The most active ethyl acetate fraction was selected for chromatographic separation to isolate six phlorotannins: phloroglucinol (1), dioxinodehydroeckol (2), eckol (3), phlorofucofuroeckol A (4), dieckol (5), and 7-phloroeckol (6). Compounds 3–6 showed potent inhibitory activity against Cu2+-induced LDL oxidation as compared with probucol, a well-known clinical therapeutic agent for hypercholesterolemia. Moreover, when compound 5 (at levels of 9 and 4.5 μM) was used in combination with probucol (4.5 μM), they additively inhibited Cu2+-induced LDL oxidation. In addition, 3–5 significantly prolonged the lag time of conjugated diene formation at 10 μM. These results suggest that the potent antiatherosclerotic effects of E. stolonifera and its isolated phlorotannins may be partly attributed to the inhibition of Cu2+-induced LDL oxidation and conjugated diene formation.

Benzophenones and xanthones from Garcinia cantleyana var. cantleyana and their inhibitory activities on human low-density lipoprotein oxidation and platelet aggregation

Three benzophenones, 2,6,3′,5′-tetrahydroxybenzophenone (1), 3,4,5,3′,5′-pentahydroxybenzophenone (3) and 3,5,3′,5′-tetrahydroxy-4-methoxybenzophenone (4), as well as a xanthone, 1,3,6-trihydroxy-5-methoxy-7-(3′-methyl-2′-oxo-but-3′-enyl)xanthone (9), were isolated from the twigs of Garcinia cantleyana var. cantleyana. Eight known compounds, 3,4,5,3′-tetrahydroxy benzophenone (2), 1,3,5-trihydroxyxanthone (5), 1,3,8-trihydroxyxanthone (6), 2,4,7-trihydroxyxanthone (7), 1,3,5,7-tetrahydroxyxanthone (8), quercetin, glutin-5-en-3β-ol and friedelin were also isolated. The structures of the compounds were elucidated by spectroscopic methods. The compounds were investigated for their ability to inhibit low-density lipoprotein (LDL) oxidation and platelet aggregation in human whole blood in vitro. Most of the compounds showed strong antioxidant activity with compound 8 showing the highest inhibition with an IC50 value of 0.5μM, comparable to that of probucol. Among the compounds tested, only compound 4 exhibited strong inhibitory activity against platelet aggregation induced by arachidonic acid (AA), adenosine diphosphate (ADP) and collagen. Compounds 3, 5 and 8 showed selective inhibitory activity on platelet aggregation induced by ADP.

The isolation and identification of two compounds with predominant radical scavenging activity in hempseed (seed of Cannabis sativa L.)

Forty samples were extracted from defatted kernels and hulls of two varieties of hempseed (Bama and Yunma No. 1) using 10 different polar solvent systems. The radical scavenging capacity of the extracts was evaluated using 2,2-diphenyl-1-pikrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays and the total phenolic content was determined by Folin-Ciocalteu's phenol reagent. The correlation analysis indicated that the antioxidants in hempseed belonged to phenolic and DPPH() assay was suitable for evaluating the radical scavenging activity. Two compounds, with predominant antiradical activity, were isolated in 60% ethanol extract of hempseed hull using macroporous resin absorption, LH-20 gel chromatography, and high performance liquid chromatography methods, which were identified as N-trans-caffeoyltyramine and cannabisin B by high-resolution mass spectra, nuclear magnetic resonance spectra, and ultraviolet data. The two compounds exhibited significant high DPPH() scavenging activity and protective effect against in vitro oxidation of human low-density lipoprotein compared with extracts from flaxseed, grape seed, and soybean. This suggests that hempseed hull extract is a potential source of natural antioxidants, which could be added to dietary supplements to help prevent oxidative stress.

Protective effect of fresh and processed Jalapeño and Serrano peppers against food lipid and human LDL cholesterol oxidation

The potential of fresh and processed Jalapeño and Serrano pepper extracts for the control of lipid oxidation in several systems was investigated. The assays used included β-carotene/linoleate model system, striped corn oil, ground pork meat and Cu(II)-induced oxidation of human low-density lipoprotein (LDL). The results showed that all extracts were effective in at least one system, although their potency varied. Extracts of smoked Jalapeño were particularly effective. The lipid-protective effect of the extracts was mainly due to the presence of phenolic compounds, but other phytochemicals, such as chlorophylls were also involved.

Differential Effects of Grape (Vitis vinifera) Skin Polyphenolics on Human Platelet Aggregation and Low-Density Lipoprotein Oxidation

Antioxidant and antiplatelet properties of grape products are thought to be responsible for observed antiatherosclerotic effects. Diverse classes of phenolics are derived from the seed and skin (GSK) of grapes. The relative contributions of the classes of phenolics to observed properties of grape products are unknown. In this paper, GSK fractions were used to examine effects on platelet aggregation, low-density lipoprotein (LDL) oxidation in vitro, and relative binding of phenolics to LDL. GSK was separated into six fractions (fractions 1-6), and primary phenolics were characterized using high-performance liquid chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Fractions 4, 5, and 6, enriched in polygalloyl polyflavan-3-ols (PGPFs) with 3-6, 4-8, and 6-15 degrees of polymerization, respectively, inhibited platelet aggregation. Fractions 1-3, containing various amounts of oligosaccharides, hydroxycinnamic acids, anthocyanins, flavanols, and low molecular weight PGPFs, significantly increased platelet aggregation. Fractions 4-6 were most effective in binding LDL and inhibiting LDL oxidation. Fractions 5 and 6 exhibited the greatest inhibition of platelet aggregation and LDL oxidation, suggesting that polymeric PGPFs are responsible for the beneficial effects of grape products. Conversely, phenolics in fractions 1-3 may reduce the net biological potency of the grape products and have undesirable effects on cardiovascular disease risk factors.

Effects of selected medicinal plants on human low-density lipoprotein oxidation, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radicals and human platelet aggregation

The effects of the methanol extracts of 20 selected medicinal plants on free radical scavenging capacity, human low-density lipoprotein (LDL) peroxidation and platelet aggregation were investigated. The antioxidant activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity and thiobarbituric acid reactive substances (TBARS) assay with LDL as the oxidation substrate. The antiplatelet activity in human whole blood was investigated using an electrical impedance method. The total phenolic contents (TPC) of the extracts were determined by the Folin-Ciocalteau method. Among the extracts, Phyllanthus amarus and Labisia pumila var. alata possessed potent radical scavenging activity with IC50 values of 3.4 and 5.7 µg/ml, respectively. The extracts ofZingiber officinale, Curcuma xanthorrhiza and Curcuma domestica showed strong inhibition of LDL peroxidation with IC50 values ranging from 0.1 to 0.9 µg/ml. The extract of Z. officinalewas the most effective sample against platelet aggregation caused by arachidonic acid (AA) and adenosine diphosphate (ADP) with IC50 values of 10.9 and 7.7 µg/ml, respectively. The Pearson correlation analysis indicated that the TPC of the extracts showed significant positive correlations with DPPH scavenging activity (r = 0.846) and LDL antioxidant activity (r= 0.639) and moderate positive correlations with antiplatelet activities (AA-induced, r = 0.51; ADP-induced, r = 0.40; collagen-induced, r = 0.44). The antioxidant and antiplatelet activities of the plant extracts could partly be due to their TPC. Key words: Medicinal plants, LDL antioxidant activity, antiplatelet activity, DPPH scavenging capacity, total phenolic contents.

Does lycopene offer human LDL any protection against myeloperoxidase activity?

Lycopene is a lipophilic antioxidant that is largely transported in human blood by Low Density Lipoproteins (LDL). One of the early events in the aetiology of atherosclerosis is thought to be the oxidation of LDL. Myeloperoxidase an enzyme secreted by neutrophils and macrophages is thought to oxidise human LDL particles. In this study, isolated human LDL was challenged with myeloperoxidase or copper, and the LDL was screened for lipoperoxidation and oxidation of apolipoprotein B100, depletion of lycopene and oxidation of cholesterol. Myeloperoxidase induced oxidation of LDL through direct interaction with apolipoprotein B100. No lipoperoxidation was observed following myeloperoxidase treatment; however, 7-ketocholesterol was detected indicating the products of myeloperoxidase interact with the surface of the LDL particles. Lycopene does react with the products of myeloperoxidase in solvent, but played no role in protecting against enzyme derived oxidation of human LDL.

The inhibiting activity of areca inflorescence extracts on human low density lipoprotein oxidation induced by cupric ion

The oxidative modification of human low density lipoprotein (LDL) plays a significant role in atherosclerosis. In this study, the inhibiting activity of areca inflorescence extracts (AIEs) on LDL oxidation was investigated by an in vitro study with Trolox as the standard antioxidant. The kinetics of LDL oxidation, thiobarbituric acid reactive substances assay, ferric-reducing antioxidant power assay and copper chelation assay were also evaluated to assess the antioxidant activities of AIEs, and the results revealed that AIEs could delay the lag time and inhibit the formation of malondialdehyde in the process of LDL peroxidation induced by Cu2+. The boiled water extract displayed the highest antioxidant activity compared with the ambient water extract and ethanol extract. The total phenolic contents and phenolic components of AIEs were also measured by high performance liquid chromatography method. Epicatechin, gallic acid and coumalic acid were the primary phenolic acids in AIEs.

Potential Health Implication of in Vitro Human Low-Density Lipoprotein–Vitamin E Oxidation Modulation by Polyphenols Derived from Côte d’Ivoire’s Oil Palm Species

Antioxidant activities of polyphenolic compounds extracted (PPEs) from ripe fruits of oil palms are investigated by studying their in vitro effects on human low-density lipoprotein (LDL) oxidation. Four oil palm species ( Elaeis guineensis ) are issued from the National Centre of Agronomic Research of Côte d'Ivoire, of which two are parental varieties (HP1 and HP2), while the other two are crossing varieties (HP3 and HP4). The main identified compounds were rutin (HP3 and HP4) and caffeic and chlorogenic (5-caffeoyl quinic) acids (HP1, HP3, and HP4). The highest total phenolic content was found for HP4, while it was significantly lower for HP2. Antioxidative effects were monitored by Cu(2+)- or 2,2'-azobis(2-amidinopropane) hydrochloride (AAPH)-induced generation of conjugated dienes (lag time and oxidation rate). The highest PPE specific antioxidant activity (SAA) values were obtained with crossing varieties (HP3 and HP4) in the copper-oxidation assay. In the AAPH-oxidation assay, SAA values were comparable for all four varieties. PPEs were effective at preventing LDL-vitamin E depletion in vitro. They could exert direct beneficial antioxidant effects on vitamin E and other antioxidants contained in food and beverages in vivo, within the gastrointestinal (GI) tract. These data could also be of particular importance for a healthier nutrition or the management of chronic diseases by a polyphenol-rich diet.

Composition of Flavonoids and Phenolic Acids in Lychee (Litchi Chinensis Sonn.) Flower Extracts and Their Antioxidant Capacities Estimated with Human LDL, Erythrocyte, and Blood Models

Lychee (Litchi chinensis Sonn.) flower is a major nectar source in Taiwan. Antioxidant activities of acetone, ethanol, and hot-water extracts of the flower were estimated through three biochemical models: inhibition of Cu(2+) -induced oxidation of human low-density lipoprotein, scavenging ability of oxygen radicals in human blood, and inhibition of human erythrocyte hemolysis induced by peroxyl radicals. Composition and content of flavonoids and phenolic acids in these extracts were also determined by high-performance liquid chromatography. Results showed that antioxidant effects of all test models as well as contents of flavonoids and phenolic acids for the lychee flower extracts were in the order: acetone extract > ethanol extract > hot-water extract. Gentistic acid and epicatechin were the major phenolic acid and flavonoid in the extracts, respectively.

The Inhibitory Effects of Fucoidans from <i>Laminaria japonica</i> on Oxidation of Human Low-Density Lipoproteins

Five Fucoidan fractions from Laminaria japonica with different sulfate content and molecular weight were obtained by anion-exchange chromatography and mild acid hydrolysis. Their antioxidant effects on azo radicals AAPH- and AMVN -induced oxidation of human low-density lipoprotein (LDL) were evaluated by monitoring cholesteryl ester hydroperoxides (CHE-OOH) formation kinetics by HPLC analysis. Two antioxidants probucol and Vc were used as comparison. Fucoidan F-C with low molecular weight 2,000-8,000 and sulfate content 24.3% had much stronger protective antioxidant effects than other four Fucoidan fractions on both hydrophilic radical AAPH and lipophilic radical AMVN induced LDL oxidation. In AMVN induced LDL oxidation system, the other four Fucoidan fractions with molecular weight of 200,000 and 20,000 were able to modify the kinetics of LDL oxidation with the similar efficiency. However, the highly sulfated Fucoidan fraction L-B with molecular weight 20,000 was completely ineffective in protecting LDL from AAPH induced oxidation, whilst it effectively suppressed the oxidation of LDL induced by AMVN. The different protective antioxidant effects of Fucoidans on AAPH and AMVN induced LDL oxidation were due to their various structures and properties, and their capacities to interact with the different sites Fucoidans reacted on LDL.

Effect of fermentation time on antioxidative activities of Ganoderma lucidum broth using leguminous plants as part of the liquid fermentation medium

Oxidative damage plays an important role in the pathology of human diseases. Ganoderma lucidum, a medicinal fungus, has been used for thousands of years in traditional Oriental medicine. It is reported to have antioxidant functions such as inhibition of lipid peroxidation. The objective of the present study was to investigate the effect of fermentation time on the antioxidative activities of G. lucidum broth filtrate using leguminous plants as part of the liquid fermentation medium. Inhibition of Cu2+-induced oxidation of human low-density lipoprotein (LDL), DPPH radical-scavenging activity, total phenolic compounds, isoflavones and protocatechuic acid were measured to evaluate the antioxidant activity of G. lucidum fermentation broth filtrate. Our results showed that black soybean and Astragalus membranaceus improved the antioxidant activity of the G. lucidum fermentation broth filtrate. Protocatechuic acid was identified by LC–MS as the antioxidant compounds whose relative potency of inhibiting LDL oxidation to Trolox is 1.55. Protocatechuic acid showed positive correlation with the antioxidant activity of the fermentation broth filtrate while isoflavones did not contribute to antioxidant activity.

The synergistic effect of daidzein and α-tocopherol or ascorbic acid on microsome and LDL oxidation

Isoflavone daidzein brings potential health benefits. Its antioxidant properties are considered to be responsible in part for its protective effects. We investigated the antioxidant effects of daidzein and its interactive effects with<br />α-tocopherol or ascorbic acid on Fe<sup>2+/</sup>ascorbate-induced oxidation of rat liver microsomes and copper-induced human low-density lipoprotein (LDL) oxidation. Although the inhibitory effect of daidzein on lipid peroxidation in microsome was weak, it effectively prevented LDL against oxidative modification by prolonging the lag time, decreasing the propagating rate, and suppressing malonaldehyde (MDA) and carbonyls formation. When daidzein was combined with α-tocopherol in microsomes oxidation and with ascorbic acid in LDL oxidation, the protection was significantly greater than the calculated additive effect of the two individual actions. Thus, daidzein can protect LDL from oxidative modification, and its combination with nutrients may be superior to the action of it alone. These results can help to get a better understanding of the interactions of different antioxidants in vivo.