Oxysterols Research Articles

Low LXA4, RvD1 and RvE1 levels may be an indicator of the development of hypertension

Vascular structure and integrity are at the forefront of blood pressure regulation. However, there are many factors that affect the responses of the vessels. One of these is the inflammatory processes associated with high cholesterol and its modification. 15-lipoxygenase (15-LOX) is the critical enzyme in cholesterol oxidation, but this enzyme is also responsible for the synthesis of specialized proresoving lipid mediators (SPMs) called Lipoxin (Lxs) and Resolvin (Rvs). In this study, we determined serum LXA4, RvD1 and RvE1 levels in newly diagnosed hypertension (HT) and normotension (NT) cases. We evaluated how the presence of hypercholesterolemia (HC) in the follow-up changes the levels of these SPMs. We found that the three SPMs we measured decreased significantly in the presence of HC. In addition, we found a negative and significant correlation with systolic blood pressure and total cholesterol levels for the three SPMs. In conclusion, HT and HC are independent risk factors for cardiovascular death. However, the presence of HC may be an important factor for the development of HT. Increasing cholesterol levels may cause 15-LOX to shift towards LDL oxidation, thus leading to inflammation. This situation may negatively affect the vascular functions in the regulation of blood pressure. Serum LXA4, RvD1 and RvE1 measurements may provide clues that represent a shift of 15-LOX enzyme activity towards cholesterol.

Oxysterols in stored powders as potential health hazards

Cholesterol oxidation products (COPs) have greater biological activity than cholesterol itself. Oxysterols reduce the nutritional value of foods and exhibit a wide range of biological activity, including pro-oxidant, carcinogenic, and cytotoxic properties. The most commonly detected oxysterols in foods are 7α-HC, 7β-HC, a product of their dehydrogenation 7-KC and α-CE, β-CE. The main dietary sources of oxysterols are eggs and egg-derived products, thermally processed milk and milk-based products, fried meat. This study aimed to measure the amount of cholesterol oxidation products in milk powder, egg powder and milk–egg powder during 24 months of storage. The changes in the selected oxysterols (determined by gas chromatography) were recorded. In milk powder, after the production process, the amount of cholesterol was 0.2 g 100 g−1 fat and in egg powder it was 3.4 g 100 g−1. After 6 months of storage, the dominant oxysterol in milk and egg powder was 7α-HC and in milk–egg powder it was 7-KC. After the storage period, oxysterols in powdered milk reached 1.81% of total cholesterol. The most stable cholesterol was in the milk–egg mixture and its oxidation was the slowest. This study showed the presence of COPs in milk powder, egg powder and milk–egg powder and the effect of storage on cholesterol oxidation.

Site of Cholesterol Oxidation Impacts Its Localization and Domain Formation in the Neuronal Plasma Membrane.

Cholesterol is integral to the structure of mammalian cell membranes. Oxidation of cholesterol alters how it behaves in the membrane and influences the membrane biophysical properties. Elevated levels of oxidized cholesterol are associated with neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, and Huntington's disease. Previous work has investigated the impact of oxidized cholesterol in the context of simple model membrane systems. However, there is a growing body of literature that shows that complex membranes possessing physiological phospholipid distributions have different properties from those of binary or trinary model membranes. In the current work, the impact of oxidized cholesterol on the biophysical properties of a complex neuronal plasma membrane is investigated using coarse-grained Martini molecular dynamics simulations. Comparison of the native neuronal membrane to neuronal membranes containing 10% tail-oxidized or 10% head-oxidized cholesterol shows that the site of oxidization changes the behavior of the oxidized cholesterol in the membrane. Furthermore, species-specific domain formation is observed between each oxidized cholesterol and minor lipid classes. Although both tail-oxidized and head-oxidized cholesterols modulate the biophysical properties of the membrane, smaller changes are observed in the complex neuronal membrane than seen in the previous work on simple binary or trinary model membranes. This work highlights the presence of compensatory effects of lipid diversity in the complex neuronal membrane. Overall, this study improves our molecular-level understanding of the effects of oxidized cholesterol on the properties of neuronal tissue and emphasizes the importance of studying membranes with realistic lipid compositions.

Back Cover: Dietary Sterols and Sterol Oxidation Products on Atherosclerosis: An Insight Provided by Liver Proteomic and Lipidomic

Mol. Nutr. Food Res. 2021, 65, 202100516 DOI: 10.1002/mnfr.202100516 Dietary phytosterols and sterol oxidation products exhibit significant different effects on atherosclerosis, and changes in liver lipid metabolism may provide another insight for their effects. As revealed by lipidomes and proteome in this report, besides cholesterol, phytosterols and sterol oxidation products also greatly affected glycerophospholipids & glycerolipids metabolism and sphingolipids metabolism. This is reported by Xuan Yang, Baiyi Lu, and co-workers in article number 2100516.

Sterols and Sterol Oxidation Products: Effect of Dietary Intake on Tissue Distribution in ApoE-Deficient Mice.

Sterols and sterol oxidation products (SOPs) are well-known dietary factors influencing atherosclerosis; however, their distribution in vivo after dietary sterol/SOP intake is still unclear. Here, we investigated the tissue distribution of sterols and SOPs in ApoE-/- mice after dietary exposure to diets supplemented with phytosterols (PS), phytosterol oxidation products (POPs), or cholesterol oxidation products (COPs). The results showed that PS intake reduced cholesterol in serum and the liver but increased cholesterol in the brain. PS intake increased the levels of PS in vivo and the levels of 7-keto- and triol-POPs in serum and the liver. COP intake elevated the level of all COPs in serum but did not change the 7-keto-cholesterol level in the liver and brain. All POPs in serum and parts of POPs in the liver and brain increased after dietary POP exposure. Our study indicated that dietary PS and SOPs accumulated in vivo with varying degrees and influenced cerebral cholesterol metabolism.

Dietary Sterols and Sterol Oxidation Products on Atherosclerosis: An Insight Provided by Liver Proteomic and Lipidomic

The development of atherosclerosis is closely associated with disorder of lipid metabolism. Dietary sterols and their oxidation products play a role in the pathogenesis of atherosclerosis. However, their effects on liver lipid metabolism during the atherosclerosis remain unknown. Here, we apply lipidomic and proteomic analysis on liver of ApoE-/- mice feed with phytosterols, cholesterol oxidation products (COPs), or phytosterol oxidation products (POPs) to profile lipid species and reveal the underlying mechanism. Dietary exposure of phytosterols, COPs, and POPs all reduce the accumulation of liver triglyceride (TG), but COPs and POPs accelerate the fibrosis of liver. Lipidomic analysis reveals that phytosterols mainly decrease the levels of phosphatidylinositol (PI), while COPs and POPs both increase the level of digalactosyldiacylglycerol (DGDG) and reduce TG with long-chain polyunsaturated fatty acids. Besides, COPs up-regulated levels of lipids associate with atherosclerosis risk, such as phosphatidylcholines (PC), phosphatidylethanolamine (PE) and ceramides (Cer). POPs down-regulate the level of acyl carnitine (AcCa). Furthermore, proteomic analysis shows that COPs promote oxidative phosphorylation and POPs inhibit the beta oxidation of fatty acids. This study reveals that phytosterols, COPs, and POPs differently change the composition and metabolism of glycerophospholipids, sphingolipids, and glycerolipids in liver of ApoE-/- mice.

Sensitive electrochemical detection of cholesterol using a portable paper sensor based on the synergistic effect of cholesterol oxidase and nanoporous gold

As a crucial biomarker for some diseases, the determination of cholesterol in human serum is of great significance for the diagnosis and prevention of these diseases. Hence, a portable cholesterol detection method is necessary for clinical and domestic applications. Here, a portable paper sensor was designed for cholesterol detection by modifying screen-printed electrode (SPE) with nanoporous gold (NPG). To achieve the reliable cholesterol detection, a synergistic strategy was proposed based on the oxidation of cholesterol by cholesterol oxidase (ChOx) and the reduction of oxidation product (H2O2) by NPG. Compared to existing electrochemical sensors, the resulting paper sensor exhibited a wider linear response in a range from 50 μM to 6 mM as well as a higher sensitivity of 32.68 μA mM−1 cm−2 with a lower detection limit of 8.36 μM. Moreover, the portable paper sensor presented strong anti-interference capability and stability in the detection of cholesterol in human serum, and the data detected by the portable paper sensor were consistent with that obtained by an automatic biochemical analyzer. These unique performances confirmed that the proposed paper sensor was a sensitive, reliable, and portable cholesterol detection method, making it a good choice for cholesterol detection.

Effect of CLA supplementation on factors related to vascular dysfunction in arteries of orchidectomized rats

The vascular endothelium is a monolayer of flat epithelial cells located between the circulating blood and the underlying connective tissue. It conveys key functions that when impaired, lead to endothelial dysfunction. This condition is responsible for the pathogenesis of vascular diseases. The cardioprotective effect of sex hormones is widely known; hence, a murine orchidectomized model has been employed to study the effects caused by their deficiency. In the search for approaches to maintain vascular health, the effect of dietary fatty acids as CLA on cardiovascular diseases has been studied. Some proven beneficial properties of CLA are antioxidant, antiatherogenic and anti-inflammatory. Our objective was to evaluate the effect of a diet supplemented with 1.8 % (w/w) of CLA, administered during eight weeks, on the amount of cholesterol oxidation products (COPs) produced by orchidectomy and on factors related to vascular dysfunction in the aorta and the mesenteric arteries. The diet with CLA prevented the increase in prostanoids formation and maintained the normal physiological conditions of NO and antioxidant activity. In addition, it prevented the increase in cholesterol and COPs at the vascular wall. CLA-supplemented diet prevented the orchidectomy-induced alterations on prostanoids, NO and COPs and also improved the antioxidant activity. These findings could contribute to understand the mechanisms of actions of CLA involved in the prevention of cardiovascular diseases.

The use of lemon juice and its role on polyunsaturated fatty acids and cholesterol oxides formation in thermally prepared sardines

Lemon juice is used to enhance the flavor of dishes prepared with fish, which is more prone to oxidation than other flesh food. This study evaluated the impact of treating sardine fillets (Sardina pilchardus) with Tahiti lemon juice and submit them to different domestic cooking methods (air fryer, grill, and deep-fat fryer), regarding the oxidation of unsaturated fatty acids and cholesterol. The juice extract was characterized by its antioxidant capacity and bioactive constituents by UHPLC-MS. Cooking caused the degradation of polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic (EPA, C20:5) and docosahexaenoic acids (DHA, C22:6), and increased the cholesterol oxides level. However, for most treatments, degradation of PUFAs and formation of cholesterol oxides were more pronounced in samples treated with juice. Lemon juice induced cholesterol oxidation, mainly in air-fried sardines where the total content of cholesterol oxides increased from 700 ± 4 (control) to 1333 ± 5 μg/g (10 % lemon juice, in dry basis). Thus, the overall data suggest that potential antioxidant compounds may have degraded during heating and the acid environment may have induced oxidative reactions.

Multiple Targets for Oxysterols in Their Regulation of the Immune System.

Oxysterols, or cholesterol oxidation products, are naturally occurring lipids which regulate the physiology of cells, including those of the immune system. In contrast to effects that are mediated through nuclear receptors or by epigenetic mechanism, which take tens of minutes to occur, changes in the activities of cell-surface receptors caused by oxysterols can be extremely rapid, often taking place within subsecond timescales. Such cell-surface receptor effects of oxysterols allow for the regulation of fast cellular processes, such as motility, secretion and endocytosis. These cellular processes play critical roles in both the innate and adaptive immune systems. This review will survey the two broad classes of cell-surface receptors for oxysterols (G-protein coupled receptors (GPCRs) and ion channels), the mechanisms by which cholesterol oxidation products act on them, and their presence and functions in the different cell types of the immune system. Overall, this review will highlight the potential of oxysterols, synthetic derivatives and their receptors for physiological and therapeutic modulation of the immune system.

Cyclodextrin dimers: A pharmaceutical engineering approach to the therapeutic extraction of toxic oxysterols

Background and Aims: Underdog Pharmaceuticals is dedicated to reversing atherosclerosis by removing 7-ketocholesterol (7KC) from aged cells and tissues. Underdog believes that the next frontier in medicine lies in attacking the aspects of aging that drive age-related disease such as atherosclerosis. 7KC is a toxic oxysterol produced by the non-enzymatic oxidation of cholesterol that accumulates in cells and tissues causing dysfunction leading to disease. Cyclodextrins (CDs) are cyclic carbohydrates typically used to solubilize small, hydrophobic molecules in drug delivery.

Reductive Power Generated by Mycobacterium leprae Through Cholesterol Oxidation Contributes to Lipid and ATP Synthesis.

Upon infection, Mycobacterium leprae, an obligate intracellular bacillus, induces accumulation of cholesterol-enriched lipid droplets (LDs) in Schwann cells (SCs). LDs are promptly recruited to M. leprae-containing phagosomes, and inhibition of this process decreases bacterial survival, suggesting that LD recruitment constitutes a mechanism by which host-derived lipids are delivered to intracellular M. leprae. We previously demonstrated that M. leprae has preserved only the capacity to oxidize cholesterol to cholestenone, the first step of the normal cholesterol catabolic pathway. In this study we investigated the biochemical relevance of cholesterol oxidation on bacterial pathogenesis in SCs. Firstly, we showed that M. leprae increases the uptake of LDL-cholesterol by infected SCs. Moreover, fluorescence microscopy analysis revealed a close association between M. leprae and the internalized LDL-cholesterol within the host cell. By using Mycobacterium smegmatis mutant strains complemented with M. leprae genes, we demonstrated that ml1942 coding for 3β-hydroxysteroid dehydrogenase (3β-HSD), but not ml0389 originally annotated as cholesterol oxidase (ChoD), was responsible for the cholesterol oxidation activity detected in M. leprae. The 3β-HSD activity generates the electron donors NADH and NADPH that, respectively, fuel the M. leprae respiratory chain and provide reductive power for the biosynthesis of the dominant bacterial cell wall lipids phthiocerol dimycocerosate (PDIM) and phenolic glycolipid (PGL)-I. Inhibition of M. leprae 3β-HSD activity with the 17β-[N-(2,5-di-t-butylphenyl)carbamoyl]-6-azaandrost-4-en-3one (compound 1), decreased bacterial intracellular survival in SCs. In conclusion, our findings confirm the accumulation of cholesterol in infected SCs and its potential delivery to the intracellular bacterium. Furthermore, we provide strong evidence that cholesterol oxidation is an essential catabolic pathway for M. leprae pathogenicity and point to 3β-HSD as a prime drug target that may be used in combination with current multidrug regimens to shorten leprosy treatment and ameliorate nerve damage.

Evaluation of the subcapsular technique for primary closure castration in donkeys (Equus asinus)

This study described the subcapsular technique for primary closure castration in donkeys with special regard to its efficiency and welfare impacts. The study was conducted on twelve adult male donkeys, allocated randomly into two groups; subcapsular castration (SC) and open castration (OC) groups, whether the donkeys were subjected to surgical castration either by subcapsular or open castration techniques, respectively. Testosterone, cortisol, lactate, glucose, total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), triglyceride (TG), and nitric oxide (NO) were measured before and after castration. Pain-associated behavioral activities were recorded post-castration. The SC was successfully performed in donkeys through a single paramedian scrotal incision. The SC was efficient as OC in reducing testosterone levels. The pain score decreased in the SC compared to the OC over time. The SC was an efficient and reliable technique for primary closure castration in donkeys with minimal postoperative complications and care and good cosmetic, physiological, and behavioral outcomes. It can be an alternative to other castration techniques in equines.

Atherogenic oxoaldehyde of cholesterol induces innate immune response in monocytes and macrophages.

Cholesterol oxidation product, 3β-hydroxy-5-oxo-5,6-secocholestan-6-al (cholesterol 5,6-secosterol, ChSeco or Atheronal-A), formed at inflammatory sites, has been shown to promote monocyte differentiation into macrophages and cause elevated expression of macrophage scavenger receptors. Since inflammation is a key event at all stages of atherosclerotic plaque formation, the pro-inflammatory actions of ChSeco in human THP-1 monocytes and mouse J774 macrophages were investigated in the present study by employing ELISA, qRT-PCR, and functional assays. An increase in the secretion of interleukin-8 and platelet-derived growth factor (PDGF) isoform AA and, to a limited extent, PDGF isoform BB was observed into the culture medium of THP-1 monocytes exposed to ChSeco. However, no changes were seen in the secretion of tumor necrosis factor-alpha. In J774 macrophages treated with ChSeco, there was an upregulation of gene expression of several pro-inflammatory cytokines and their receptors. Concomitantly, there was down-regulation of gene expression of interleukin-1ß, interleukin-10, and lymphotoxin-beta. An increase in the release of interleukin-18 and chemokine (C-C motif) ligand-20 from J774 macrophages (which corroborated well with their gene expression profiles) and increased binding of THP-1 monocytes to ChSeco-exposed human aortic endothelial cells were observed. The results of the present study, for the first time, demonstrate the pro-inflammatory action of ChSeco and suggest the underlying pro-atherogenic mechanisms. These could be mediated through enhanced monocyte recruitment into the sub-endothelial space and subsequent proliferation of smooth muscle cells under the influence of monocyte-derived PDGF.

7-Ketocholesterol: Effects on viral infections and hypothetical contribution in COVID-19

7-Ketocholesterol, which is one of the earliest cholesterol oxidization products identified, is essentially formed by the auto-oxidation of cholesterol. In the body, 7-ketocholesterol is both provided by food and produced endogenously. This pro-oxidant and pro-inflammatory molecule, which can activate apoptosis and autophagy at high concentrations, is an abundant component of oxidized Low Density Lipoproteins. 7-Ketocholesterol appears to significantly contribute to the development of age-related diseases (cardiovascular diseases, age-related macular degeneration, and Alzheimer's disease), chronic inflammatory bowel diseases and to certain cancers. Recent studies have also shown that 7-ketocholesterol has anti-viral activities, including on SARS-CoV-2, which are, however, lower than those of oxysterols resulting from the oxidation of cholesterol on the side chain. Furthermore, 7-ketocholesterol is increased in the serum of moderately and severely affected COVID-19 patients. In the case of COVID-19, it can be assumed that the antiviral activity of 7-ketocholesterol could be counterbalanced by its toxic effects, including pro-oxidant, pro-inflammatory and pro-coagulant activities that might promote the induction of cell death in alveolar cells. It is therefore suggested that this oxysterol might be involved in the pathophysiology of COVID-19 by contributing to the acute respiratory distress syndrome and promoting a deleterious, even fatal outcome. Thus, 7-ketocholesterol could possibly constitute a lipid biomarker of COVID-19 outcome and counteracting its toxic effects with adjuvant therapies might have beneficial effects in COVID-19 patients.

Mestranol moieties clicked to Zn(II)phthalocyanine for controllable photosensitized oxidation of cholesterol

Four mestranol moieties were chemically linked to Zn(II) phthalocyanine (4) by cycloaddition “Click” reaction using a tetra-azidoethoxy substituted Zn(II)-phthalocyanine (3). The alkyl-azido coupling reaction was realized between azido groups of 3 and alkyl group of mestranol. The alkylation reaction was carried out to obtain cationic Zn(II) phthalocyanine derivative (5). The new compounds were chemically characterized by the known analytical methods. The absorption and fluorescence properties were studied in comparison. The absorption maxima of phthalocyanines 3, 4 and 5 were recorded at approx. shifts of 8 - 12 nm in the far- red region (680 - 684 nm) and the fluorescence maxima (692 - 693 nm) as compared to unsubstituted ZnPc (672 nm, 680 nm) in DMSO. The studies of singlet oxygen generation of 3, 4 and 5 showed relatively high values such as 0.52 for 3; 0.51 for 4 and 0.46 for 5. The fluorescence lifetime of 3.15 ns (3), 3.25 ns (4) and 3.46 ns (5) were determined with lower than the value than for the used standard ZnPc (3.99 ns). The high photo stability was observed for compounds 3, 4 and 5. In addition, the photosensitized oxidation of cholesterol was compared for 3 and 4 with much lower values of oxidation potential than for unsubstituted ZnPc which suggests that the substitution groups influenced on the photooxidation index of the target molecule.

The Controversial Role of 24-S-Hydroxycholesterol in Alzheimer's Disease.

The development of Alzheimer’s disease (AD) is influenced by several events, among which the dysregulation of cholesterol metabolism in the brain plays a major role. Maintenance of brain cholesterol homeostasis is essential for neuronal functioning and brain development. To maintain the steady-state level, excess brain cholesterol is converted into the more hydrophilic metabolite 24-S-hydroxycholesterol (24-OHC), also called cerebrosterol, by the neuron-specific enzyme CYP46A1. A growing bulk of evidence suggests that cholesterol oxidation products, named oxysterols, are the link connecting altered cholesterol metabolism to AD. It has been shown that the levels of some oxysterols, including 27-hydroxycholesterol, 7β-hydroxycholesterol and 7-ketocholesterol, significantly increase in AD brains contributing to disease progression. In contrast, 24-OHC levels decrease, likely due to neuronal loss. Among the different brain oxysterols, 24-OHC is certainly the one whose role is most controversial. It is the dominant oxysterol in the brain and evidence shows that it represents a signaling molecule of great importance for brain function. However, numerous studies highlighted the potential role of 24-OHC in favoring AD development, since it promotes neuroinflammation, amyloid β (Aβ) peptide production, oxidative stress and cell death. In parallel, 24-OHC has been shown to exert several beneficial effects against AD progression, such as preventing tau hyperphosphorylation and Aβ production. In this review we focus on the current knowledge of the controversial role of 24-OHC in AD pathogenesis, reporting a detailed overview of the findings about its levels in different AD biological samples and its noxious or neuroprotective effects in the brain. Given the relevant role of 24-OHC in AD pathophysiology, its targeting could be useful for disease prevention or slowing down its progression.

Zinc Affects Cholesterol Oxidation Products and Fatty Acids Composition in Rats' Serum.

The purpose of this work was to evaluate the effect of the nanosized or microsized zinc (Zn) particles on fatty acid profile, enzyme activity and the level of cholesterol, squalene and oxysterols in rats with breast cancer. Rats (female, n = 24) were divided into the following groups: control, and two test groups, whose diets were enriched with either Zn microparticles (342 nm) or Zn nanoparticles (99 nm). All rats were treated twice with the carcinogenic agent; 7,12-dimethylbenz[a]anthracene. In rats whose diet was enriched with zinc (especially in the form of nanoparticles), the number and sizes of tumors were lower. Diet supplementation also significantly reduced the cholesterol (p = 0.027) and COPs (cholesterol oxidation products) levels (p = 0.011) in rats serum. Enriching the diet with Zn microparticles decreased the Δ6-desaturase activity (p < 0.001). Zn influences fatty acids’ profile in rats’ serum as well as inhibiting desaturating enzymes. A reduced amount of pro-inflammatory arachidonic acid derivatives may be the expected effect.

Cholesterol-Induced M4-Like Macrophages Recruit Neutrophils and Induce NETosis.

The liver is the central organ for cholesterol synthesis and homeostasis. The effects of dietary cholesterol on hepatic injury, mainly of oxidized low-density lipoproteins (OxLDL), are not fully understood. Here, we show that the degree of cholesterol oxidation had different impacts on the global gene expression of human M2-like macrophages, with highly oxidized LDL causing the most dramatic changes. M2-like macrophages and Kupffer cells undergo M4-like polarization, decreasing the expression of important markers, such as IL10, MRC1, and CD163. These cells also displayed functional changes, with reduced phagocytic capacity, increased neutrophil recruitment, and more effective neutrophil extracellular traps (NETs) induction. Our findings provide a link between LDL oxidation and modification of peripheral and liver macrophage function.

Effects of a Combination of Extracts from Olive Fruit and Almonds Skin on Oxidative and Inflammation Markers in Hypercholesterolemic Subjects: A Randomized Controlled Trial.

Hydroxytyrosol (HT) from olives and polyphenols from almond skin (ASPs) possess cardioprotective properties. This pilot study evaluates the effect of supplementation with a combination of olive fruit and almond skin extracts on low-density lipoprotein (LDL) cholesterol oxidation, lipid homeostasis, and inflammatory parameters in adults with moderate hypercholesterolemia. A randomized, parallel, double-blind, placebo-controlled pilot study of 8 weeks was performed. The extract group (EG) received the supplement with 7.5 mg HT +210 mg ASPs, and the control group (CG) received a placebo composed of maltodextrin. Oxidized LDL (oxLDL) levels and the oxLDL/LDL ratio were lower in the EG than in the CG after 8 weeks of treatment (18.76 ± 3.91 vs. 10.34 ± 4.22, P < .001 and 0.151 ± 0.025 vs. 0.08 ± 0.023, P < .001, respectively). Interleukin-1β levels were significantly higher in the CG than in the EG at week 4 (P = .004), IL-6 was significantly higher in the CG than in the EG at week 4 (P = .049), and IL-10 was significantly increased at week 4 in both groups (P = .002 for CG and P = .001 for EG). In conclusion, daily consumption of a combination of an olive fruit extract and an almond skin extract for 8 weeks seems to protect LDL from oxidation and to prevent inflammatory status in moderately hypercholesterolemic subjects.