Comparative study between n-6, trans and n-3 fatty acids on repeated amphetamine exposure: a possible factor for the development of mania

In the last decades, foods rich in omega-3 (ω-3) fatty acids (FA) have been replaced by omega-6 (ω-6) and trans FA, which are found in processed foods. The influence of ω-6 (soybean oil - SO), trans (hydrogenated vegetable fat - HVF) and ω-3 (fish oil - FO) fatty acids on locomotor and oxidative stress (OS) parameters were studied in an animal model of mania. Rats orally fed with SO, HVF and FO for 8 weeks received daily injections of amphetamine (AMPH — 4 mg/kg/mL-ip) for the last week of oral supplementation. HVF induced hyperactivity, increased the protein carbonyl levels in the cortex and decreased the mitochondrial viability in cortex and striatum. AMPH-treatment increased the locomotion and decreased the mitochondrial viability in all groups, but its neurotoxicity was higher in the HVF group. Similarly, AMPH administration increased the protein carbonyl levels in striatum and cortex of HVF-supplemented rats. AMPH reduced the vitamin-C plasmatic levels of SO and HVF-fed rats, whereas no change was observed in the FO group. Our findings suggest that trans fatty acids increased the oxidative damage per se and exacerbated the AMPH-induced effects. The impact of trans fatty acids consumption on neuronal diseases and its consequences in brain functions must be further evaluated.     

Differential modulation by simvastatin of the metabolic pathways in the n-9, n-6 and n-3 fatty acid series, in human monocytic and hepatocytic cell lines

Statins affect the production of long chain polyunsaturated fatty acids (PUFA), both in vitro and in vivo. Various studies have shown the effects of statins on the pattern of n-6 fatty acids (FA), but limited attention has been paid to the n-3 FA. We investigated, in THP-1 and in HepG2 cells, the effects of simvastatin on the conversion of the 18C FA precursors in the n-3 and n-6 series, [1-(14)C] alpha-linolenic acid (alpha-LNA) and [1-(14)C] linoleic acid (LA) respectively, and on the metabolism of [1-(14)C] stearic acid (SA). THP-1 cells, as in the case of LA, actively converted alpha-LNA to its products, and after simvastatin treatment, the total conversion was significantly increased (from 57.2+/-7.2 to 74.3+/-8.5%, p<0.05). HepG2 cells also converted LA and alpha-LNA, but simvastatin increased significantly only the conversion of LA (9.5+/-1.9% versus 23.8+/-5.1%, p<0.02). SA conversion was similar in untreated cells (about 50%), while statin increased the production of oleic acid in HepG2, but in THP-1 cells there was a decrease. In conclusion, LA, alpha-LNA and SA are differentially metabolized in THP-1 and in HepG2 cells and their increased conversion by simvastatin is lower in HepG2 than in THP-1. These differences may reflect the distinct features of the two cell lines: monocytes, precursors of phagocytic cells, versus hepatocytes with mainly metabolic functions. Substantial differences concern also cellular FA pools: structural in THP-1 cells, and also depot, resulting in sequestering of the substrates, in HepG2. The greater n-3 FA metabolism in THP-1 cells may have favourable functional effects.     

Polyunsaturated fatty acid-based targeted nanotherapeutics to enhance the therapeutic efficacy of docetaxel

Since breast cancer is one of the most lethal malignancies, targeted strategies are urgently needed. In this study, we report the enhanced therapeutic efficacy of docetaxel (DTX) when combined with polyunsaturated fatty acids (PUFA) for effective treatment of multi-resistant breast cancers. Folic acid (FA)-conjugated PUFA-based lipid nanoparticles (FA-PLN/DTX) was developed. The physicochemical properties, in vitro uptake, in vitro cytotoxicity, and in vivo anticancer activity of FA-PLN/DTX were evaluated. FA-PLN/DTX could efficiently target and treat human breast tumor xenografts in vivo. They showed high payload carrying capacity with controlled release characteristics and selective endocytic uptake in folate receptor-overexpressing MCF-7 and MDA-MB-231 cells. PUFA synergistically improved the anticancer efficacy of DTX in both tested cancer cell lines by inducing a G2/M phase arrest and cell apoptosis. Combination of PUFA and DTX remarkably downregulated the expression levels of pro-apoptotic and anti-apoptotic markers, and blocked the phosphorylation of AKT signaling pathways. Compared to DTX alone, FA-PLN/DTX showed superior antitumor efficacy, with no signs of toxic effects in cancer xenograft animal models. We propose that PUFA could improve the therapeutic efficacy of anticancer agents in cancer therapy. Further studies are necessary to fully understand these findings and achieve clinical translation.     

Modulation of Fear Memory by Dietary Polyunsaturated Fatty Acids via Cannabinoid Receptors

Although the underlying mechanism remains unknown, several studies have suggested benefits of n-3 long-chain polyunsaturated fatty acid (PUFA) for patients with anxiety disorders. Elevated fear is thought to contribute to the pathogenesis of particular anxiety disorders. The aim of the present study was to evaluate whether the dietary n-3 to n-6 PUFA (3:6) ratio influences fear memory. For this purpose, the effects of various dietary 3:6 ratios on fear memory were examined in mice using contextual fear conditioning, and the effects of these diets on central synaptic transmission were examined to elucidate the mechanism of action of PUFA. We found that fear memory correlated negatively with dietary, serum, and brain 3:6 ratios in mice. The low fear memory in mice fed a high 3:6 ratio diet was increased by the cannabinoid CB₁ receptor antagonist rimonabant, reaching a level seen in mice fed a low 3:6 ratio diet. The agonist sensitivity of CB₁ receptor was enhanced in the basolateral nucleus of the amygdala (BLA) of mice fed a high 3:6 ratio diet, compared with that of mice fed a low 3:6 ratio diet. Similar enhancement was induced by pharmacological expulsion of cholesterol in the neuronal membrane of brain slices from mice fed a low 3:6 ratio diet. CB₁ receptor-mediated short-term synaptic plasticity was facilitated in pyramidal neurons of the BLA in mice fed a high 3:6 ratio diet. These results suggest that the ratio of n-3 to n-6 PUFA is a factor regulating fear memory via cannabinoid CB₁ receptors.     

北沙参的脂肪酸特征及产地差异性分析

目的 研究北沙参的脂肪酸特征,比较山东莱阳、河北安国、内蒙古赤峰等三个主产地的北沙参脂肪酸差异。方法 Bligh and Dyer法提取总脂;甲酯化后,利用气相色谱-质谱联用仪（GC-MS）分析;通过与脂肪酸标准品及NIST 11.0质谱数据库比对鉴定脂肪酸种类,采用面积归一法分别计算各成分的相对质量分数。结果 分别从山东莱阳、河北安国和内蒙古赤峰的北沙参药材中鉴定了17、17和18种脂肪酸;三个产地药材的优势脂肪酸种类一致,依次为C18:2 n-6c（亚油酸,49.22~63.96%）、C16:0（棕榈酸,17.43~25.33%）和C18:1 n-9c（油酸,13.85~19.44%）,均未检测到n-3型多不饱和脂肪酸（PUFA）。山东莱阳药材的多不饱和脂肪酸总量（PUFA）低于河北安国及内蒙古赤峰,但饱和脂肪酸（SFA）、 单不饱和脂肪酸（MUFA）和多不饱和脂肪酸（PUFA）三者比值最接近1:1:1。结论 不同产地北沙参的脂肪酸种类相近。亚油酸是北沙参含量优势脂肪酸,提示可以作为北沙参防治胆固醇代谢相关疾病的质量标志物候选分子。     

In vitro response of ATPase activities in tissue subcellular particle preparations to a series of mono-unsaturated C18 fatty acids

The positions of the double bond and the cis/trans configurations of six mono-unsaturated C18 fatty acids (FA) showed selectivity for inhibition and stimulation of ATPase activities of tissue homogenate fractions. The 13,000 g and 100,000 g sediments (fractions B and C respectively) of tissues homogenates were used as sources of Na+-K+ ATPase and of oligomycin-sensitive (OS) and -insensitive (OIS) Mg2+ ATPase activities. Tissue source included bovine brain and rat brain, kidney, heart and liver. Cis mono-unsaturated C18 FA caused an apparent uncoupling of mitochondrial coupling factor F1 (Mg2+ ATPase). This was indicated by the loss of oligomycin and 1,1,1-trichloro-2,2 bis (p-chlorophenyl) ethane (DDT) sensitivity in the presence of 50 microM FA (12-C18:1). Thus, a precipitous decrease in OS-Mg2+ AtPase activity of mitochondria was accompanied by an equally steep increase in OIS-Mg2+ ATPase activity. This was especially apparent in the heart tissue preparation. The uncoupling action of the FA was not observed with the trans mono-unsaturated C18 FA, Na+-K+ ATPase activity from a bovine brain nerve ending particle (B) fraction was also sensitive to 12-C18:1 FA. However, inhibitory response of Na+-K+ ATPase activity were different for OS-Mg2+ ATPase activity. The latter (OS) was not sensitive to the trans 12-C18:1, while the former (Na+-K+) was sensitive to both cis and trans forms of 12-C18:1 and inhibition appeared to be dependent on the position of the double bond.     

Lipidomics reveals the dynamics of lipid profile altered by omega-3 polyunsaturated fatty acid supplementation in healthy people

Glycerophospholipids (GPs) and sphingolipids (SPs) are important lipid components in the body and play biological functions. Omega-3 polyunsaturated fatty acids (n-3 PUFAs) are important nutrients, and their supplements are commonly used for preventing some diseases. However, the effect of n-3 PUFAs on the human glycerophospholipidome and sphingolipidome is unclear. We used targeted lipidomics to study the GP and SP profile of healthy individuals after supplementation with n-3 PUFAs for 3, 7, 14 and 21 days. Fuzzy c-means clustering was used to cluster the lipid species into six classes reflecting different changed-content patterns after n-3 PUFA supplementation. Among the species with significantly changed content, lysophospholipids were the most sensitive; their content started to increase on day 3. The content of phosphatidylserines increased at a later stage. The content of most of the phosphatidylcholines and alkylphosphatidylcholines decreased on day 21. A correlation network analysis of lipid species suggested that some enzymes involved in the metabolism of lysophospholipids and phosphatidylserines were regulated by n-3 PUFAs. Levels of creatine kinase-MB (CK-MB), urea, glucose, triglycerides and total bilirubin were altered by n-3 PUFA at 21 days. Correlation analysis revealed that the level of CK-MB was negatively correlated with those of species in lysophosphatidic acid, lysophosphatidylcholine, lysophosphatidylethanolamine and phosphatidylserine classes, which were increased by n-3 PUFA supplementation. With the analysis in this work, we demonstrated the regular pattern of n-3 PUFAs on GP and SP metabolism, which provides a pharmacological basis for n-3 PUFAs for clinical application.     

n-3多不饱和脂肪酸类药物在心血管疾病中的临床应用进展

许多临床试验表明n-3多不饱和脂肪酸(n-3 PUFAs)对于冠心病、血脂异常和心力衰竭等人群均具有保护作用,而且已有建议推荐心肌梗死后患者和高甘油三酯血症人群口服此类提纯药物。但在应用过程中,仍有一些值得临床关注的问题,如药物安全性、药物成分和用量等。笔者认为,随着对n-3 PUFAs药物的深入研究,其将有更广阔的应用前景。     

Chronic dietary changes in n-6/n-3 polyunsaturated fatty acid ratios cause developmental delay and reduce social interest in mice

Polyunsaturated fatty acids (PUFAs) are one of the main cellular building blocks, and dietary changes in PUFA composition are proposed as a potential route to influence brain development. For example, initial studies indicated that there is a relation between blood omega-6(n-6)/omega-3(n-3) PUFA ratios and neurodevelopmental disease diagnosis. To study the consequences of dietary n-6/n-3 PUFA ratio changes, we investigated the impact of a n-3 supplemented and n-3 deficient diet in developing BTBR T?+?Itpr3tf/J (BTBR) – a mouse inbred strain displaying Autism Spectrum Disorder (ASD)-like symptomatology - and control C57BL/6J mice. This study showed that pre- and postnatal changed dietary n-6/n-3 ratio intake has a major impact on blood and brain PUFA composition, and led to delayed physical development and puberty onset in both strains. The PUFA induced developmental delay did not impact adult cognitive performance, but resulted in reduced social interest, a main ASD behavioral feature. Thus, both chronic dietary n-3 PUFA supplementation and depletion may not be beneficial.     

Dietary n-6 and n-3 polyunsaturated fatty acids and colorectal carcinogenesis: results from cultured colon cells, animal models and human studies

During the past few decades, many studies have been conducted to evaluate the effects of n-6 and n-3 polyunsaturated fatty acids (PUFAs) on colorectal carcinogenesis. This report provides a brief overview of the recent studies that have been performed in cultured colon cells, animal models as well as of the population-based and short-term biomarker studies with humans. No differential effect between n-6 and n-3 PUFAs has been observed in vitro. Results from animal models indicate that n-6 PUFAs have a tumor enhancing effect, predominantly during the post-initiation phase. n-3 PUFAs may protect against colorectal carcinogenesis during both the initiation and post-initiation phase. Population-based human studies show little or no associations between n-6 or n-3 PUFA intake and colorectal cancer. Short-term biomarker studies in humans suggest though that fish oil (FO) supplementation with high amounts of n-3 PUFAs may protect against colorectal carcinogenesis and that n-6 PUFA supplementation may increase the risk.     

Dietary n-6 and n-3 polyunsaturated fatty acids and colorectal carcinogenesis: results from cultured colon cells, animal models and human studies

During the past few decades, many studies have been conducted to evaluate the effects of n-6 and n-3 polyunsaturated fatty acids (PUFAs) on colorectal carcinogenesis. This report provides a brief overview of the recent studies that have been performed in cultured colon cells, animal models as well as of the population-based and short-term biomarker studies with humans. No differential effect between n-6 and n-3 PUFAs has been observed in vitro. Results from animal models indicate that n-6 PUFAs have a tumor enhancing effect, predominantly during the post-initiation phase. n-3 PUFAs may protect against colorectal carcinogenesis during both the initiation and post-initiation phase. Population-based human studies show little or no associations between n-6 or n-3 PUFA intake and colorectal cancer. Short-term biomarker studies in humans suggest though that fish oil (FO) supplementation with high amounts of n-3 PUFAs may protect against colorectal carcinogenesis and that n-6 PUFA supplementation may increase the risk.     

Use of Fatty Acids to Explain Variability of Organochlorine Concentrations in Eggs and Plasma of Common Terns (Sterna hirundo)

We have studied the breeding parameters, organochlorine compounds (OCs) concentrations, and fatty acid (FA) composition of egg yolks (n = 47) and plasma (n = 90) of common terns (Sterna hirundo) from two colonies (Banya and Fangar) in the Ebro delta, NE Spain. Terns from Banya tend to have smaller clutch size and lower hatching success than terns from Fangar. p,p'-DDE and polychlorinated biphenyls (PCBs) concentrations were almost 2-fold higher in yolks from Banya in 1998 than from Fangar in 1999, and the percentage of n-6 PUFA was positively correlated with these contaminants. Differences between samplings in OCs concentrations in plasma were less evident, and were affected by breeding chronology. The highest OCs concentrations in yolks from Banya may be explained by two processes involving the increased deposition of n-6 PUFA: (1) higher mobilization of endogenous fat due to lower food resources, or (2) differences in the diet between colonies. Birds from Banya may have been feeding at a higher degree on discards of trawling fisheries composed of demersal and benthic fish species that are more exposed to contaminants from sediment and have lower n-3/n-6 ratio, whereas birds from Fangar would feed mostly on pelagic species of small clupeiformes that are less polluted and have higher n-3/n-6 ratios.     

Neoplastic and preneoplastic lesions induced by melamine in rat urothelium are modulated by dietary polyunsaturated fatty acids

The modulatory effects of polyunsaturated fatty acids (PUFA) on urinary tract tumorigenesis of 275 Wistar rats were evaluated by treating animals with the tumorigenic agent melamine. Rats were fed with formulae containing 6% of 4 varieties of fats: fish oil enriched in n-3 PUFA (FO), corn oil enriched in n-6 (CO), olein containing mainly n-9 oleic acid (O), and 98% stearic acid (SA), the latter two being essential (EFA)-deficient inducers. Two commercially fed control groups with (CM) and without (C) melamine were used. Animals were autopsied at 22–25 and at 36–40 weeks. Hepatic fatty acids showed that O and SA groups were EFA-deficient. Simple well differentiated hyperplasias were significantly higher in the FO lot, whereas dysplasia was increased in the CO, O and SA lots. Most of the animals fed for 36–40 weeks with the three latter formulae developed the more severe lesions. Increased urothelial proliferation was more frequent in EFA-deficient rats. The apoptosis/mitosis ratio was higher in O, SA and CO fed animals with respect to FO and chow ones. Results show that dietary PUFA modulate differentially both normal and pre-neoplastic urothelial proliferation induced by melamine. FO, rich in n-3 fatty acids, showed a strong protective effect.     

A comparative study on the effect of algal and fish oil on viability and cell proliferation of Caco-2 cells.

Polyunsaturated fatty acid (PUFA) rich micro-algal oil was tested in vitro and compared with fish oil for antiproliferative properties on cancer cells in vitro. Oils derived from Crypthecodinium cohnii, Schizochytrium sp. and Nitzschia laevis, three commercial algal oil capsules, and menhaden fish oil were used in cell viability and proliferation tests with human colon adenocarcinoma Caco-2 cells. With these tests no difference was found between algal oil and fish oil. The nonhydrolysed algal oils and fish oil showed a much lower toxic effect on cell viability, and cell proliferation in Caco-2 cells than the hydrolysed oils and the free fatty acids (FFAs). Eicosapentaenoic acid (EPA; C20:5n-3) and docosahexaenoic acid (DHA; C22:6n-3) were used as samples for comparison with the tested hydrolysed and nonhydrolysed oils. The hydrolysed samples showed comparative toxicity as the free fatty acids and no difference between algal and fish oil. Oxidative stress was shown to play a role in the antiproliferative properties of EPA and DHA, as alpha-tocopherol could partially reverse the EPA/DHA-induced effects. The results of the present study support a similar mode of action of algal oil and fish oil on cancer cells in vitro, in spite of their different PUFA content.     

Fish oil concentrate delays sensitivity to thermal nociception in mice

Fish oil has been used to alleviate pain associated with inflammatory conditions such as rheumatoid arthritis. The anti-inflammatory property of fish oil is attributed to the n-3 fatty acids docosahexaenoic acid and eicosapentaenoic acid. Contrarily, vegetable oils such as safflower oil are rich in n-6 fatty acids which are considered to be mediators of inflammation. This study investigates the effect of n-3 and n-6 fatty acids rich oils as dietary supplements on the thermally induced pain sensitivity in healthy mice. C57Bl/6J mice were fed diet containing regular fish oil, concentrated fish oil formulation (CFO) and safflower oil (SO) for 6 months. Pain sensitivity was measured by Plantar test and was correlated to the expression of acid sensing ion channels (ASICs), transient receptor potential vanilloid 1 (TRPV1) and c-fos in dorsal root ganglion cells. Significant delay in sensitivity to thermal nociception was observed in mice fed CFO compared to mice fed SO (p < 0.05). A significant diminution in expression of ion channels such as ASIC1a (64%), ASIC13 (37%) and TRPV1 (56%) coupled with reduced expression of c-fos, a marker of neuronal activation, was observed in the dorsal root ganglion cells of mice fed CFO compared to that fed SO. In conclusion, we describe here the potential of fish oil supplement in reducing sensitivity to thermal nociception in normal mice.     

Fish oil prophylaxis attenuates rotenone-induced oxidative impairments and mitochondrial dysfunctions in rat brain

Studies describing the potential of fish oil (FO) prophylaxis to render neuroprotection in experimental model/s are limited. In the present study, we have examined the propensity of FO supplements to modulate endogenous markers of oxidative stress and attenuate neurotoxicant-induced oxidative stress and mitochondrial dysfunctions in rat brain. Prepubertal male rats given FO supplements (oral, 2 and 4 mL/kg bw/day, for 30 days) showed no alterations in malondialdehyde and reactive oxygen species in brain regions. However, FO supplements significantly enhanced the GSH levels in all brain regions examined, while differential effect was discernible in the activity levels of antioxidant enzymes. Further, we investigated whether FO prophylaxis could offset Rotenone (ROT, a well known neurotoxicant) induced early oxidative stress and mitochondrial dysfunctions. While ROT elicited marked oxidative stress as evidenced by the elevated levels of malondialdehyde, hydroperoxides and protein carbonyls in the cerebellum, FO prophylaxis significantly attenuated the effect. FO prophylaxis offered varying degree of protection against ROT-induced mitochondrial dysfunctions. Collectively, our findings in the ROT model allow us to hypothesize that the prophylactic protection offered by FO may be due to its ability to enhance GSH levels, antioxidant machinery, offset protein oxidation and specific modulatory effects on brain mitochondria.     

Ozone toxicity: Effect of dietary vitamin E and polyunsaturated fatty acids

The roles of vitamin E and dietary polyunsaturated fatty acids (PUFA) in the acute toxicity of ozone have been investigated. After equilibration on test diets, the lung lipids of male CD-1 mice had an altered peroxidizability index (PI) which was effected by the content of dietary PUFA but not by vitamin E acetate. Mice fed 0 or 10.5 vitamin E acetate had the same mortality (LT50 of 29–32 days) on exposure to 1 ppm ozone, regardless of the significant differences in lung PI between groups fed high or low PUFA diets. High supplemental levels (105.0 ) of vitamin E acetate were protective and delayed the LT50 to 1 ppm ozone by an average of 15 days. The mode of action of vitamin E and the interaction with high levels of PUFA and ozone appear complex.     

Effects of n-3 polyunsaturated fatty acids on depressive symptoms,anxiety and emotional state in patients with acute myocardial infarction

BackgroundOur aim was to assess whether an early introduced n-3 polyunsaturated fatty acids (n-3 PUFA) supplementation affects depression symptoms, anxiety and emotional state in patients with acute myocardial infarction (AMI) and no history of mental disorders.MethodsFifty two patients with AMI were enrolled into the study and randomized to the study group (group P; n = 26; standard therapy + n-3 PUFA1 g daily) or the control group (group C; n = 26; standard therapy). The following psychological tests were used at the baseline (3rd day of AMI) and after one month (30 ± 1 days): Beck Depression Inventory (BDI), State-Trait Anxiety Inventory in a specific situation (STAI-S) and as a general trait (STAI-T), Emotional State Questionnaire (ESQ).ResultsThe baseline characteristics, pharmacotherapy and BDI, STAI-S/T and ESQ were similar between both groups. The mean test scores assessed for all patients (group P and C) during the one-month observation were significantly lower for BDI (p = 0.04), STAI-T (p = 0.03), STAI-S (p = 0.01) and harm/loss emotions (p = 0.005). After adjusting for age, sex, body mass index, coronary artery disease severity, ejection fraction, serum troponin level and the baseline tests results, n-3 PUFAintervention revealed additional significant decrease in BDI (p = 0.046), STAI-S (p = 0.03) and harm/loss emotions (p = 0.04).ConclusionsOur study provides novel and preliminary observations – n-3 PUFAsupplementation reveals additional decreasing effects on depressive and anxiety symptoms in early post-MI patients.     

Early modifications of fatty acid composition in plasma phospholipids,platelets and mononucleates of healthy volunteers after low doses of n-3 polyunsaturated fatty acids

Objectives Although previous data suggested that only doses of 4 g/day or higher of n-3 polyunsaturated fatty acids (PUFA) have had a beneficial effect in the prevention of atherosclerosis and cardiovascular diseases, the GISSI-Prevenzione Study in a 3-year trial showed that 1 g/day reduced total and cardiovascular mortality in over 11,000 post-infarction patients. The aim of this study was to investigate the time course and the extent of incorporation of n-3 fatty acids in plasma and blood cells after 1 g/day of n-3 PUFA, the dose effective in the GISSI-Prevenzione in comparison with higher doses.Methods Thirty-six healthy volunteers were given 1, 2 and 4 g/day of n-3 PUFA ethyl esters for 12 weeks, followed by a 4-week washout. Blood was collected at weeks 0, 1, 2, 4, 8, 12 and 16 and used for lipid profile analysis and measurement of fatty acid composition in plasma phospholipids, platelets and mononucleates.Results Total n-3 PUFA increased by 2.0-, 2.2- and 2.9-fold versus baseline after 12-week treatment with 1, 2 and 4 g respectively. A statistically significant raise of total n-3 PUFA was seen in platelets and mononucleates. Among individual n-3 PUFA, 22:5 n-3 was enriched early and dose dependently in plasma phospholipids, platelets and mononucleates; the raise of 22:6 n-3 was less marked especially in platelets and mononucleates.Conclusions One gram per day of n-3 PUFA induces fast (within 1 week) and striking changes in blood composition of PUFA that may well explain their beneficial effects against cardiovascular diseases.Delia Di Stasi and Roberto Bernasconi contributed equally to this work.