Cinta Bladé

Grape seed procyanidins improve atherosclerotic risk index and induce liver CYP7A1 and SHP expression in healthy rats

Moderate consumption of red wine reduces risk of death from cardiovascular disease. The polyphenols in red wine are ultimately responsible for this effect, exerting antiatherogenic actions through their antioxidant capacities and modulating intracellular signaling pathways and transcriptional activities. Lipoprotein metabolism is crucial in atherogenesis, and liver is the principal organ controlling lipoprotein homeostasis. This study was intended to identify the primary effects of procyanidins, the most abundant polyphenols in red wine, on both plasma lipoprotein profile and the expression of genes controlling lipoprotein homeostasis in the liver. We show that procyanidins lowered plasma triglyceride, free fatty acids, apolipoprotein B (apoB), LDL‐cholesterol and nonHDL:nonLDL‐cholesterol levels and slightly increased HDL‐cholesterol. Liver mRNA levels of small heterodimer partner (SHP), cholesterol 7α‐hydroxylase (CYP7A1), and cholesterol biosynthetic enzymes increased, whereas those of apoAII, apoCI, and apoCIII decreased. Lipoprotein lipase (LPL) mRNA levels increased in muscle and decreased in adipose tissue. In conclusion, procyanidins improve the atherosclerotic risk index in the postprandial state, inducing in the liver the overexpression of CYP7A1 (suggesting an increase of cholesterol elimination via bile acids) and SHP, a nuclear receptor emerging as a key regulator of lipid homeostasis at the transcriptional level. These results could explain, at least in part, the beneficial long‐term effects associated with moderate red wine consumption.

Hypolipidemic effects of proanthocyanidins and their underlying biochemical and molecular mechanisms.

Proanthocyanidins are the most abundant polyphenols in human diets. Epidemiological studies strongly suggest that proanthocyanidins protect against cardiovascular diseases. Despite the antioxidant and anti-inflammatory properties of these flavonoids, one of the mechanisms by which proanthocyanidins exert their cardiovascular protection is improving lipid homeostasis. Animal studies demonstrate that proanthocyanidins reduce the plasma levels of atherogenic apolipoprotein B-triglyceride-rich lipoproteins and LDL-cholesterol but increase antiatherogenic HDL-cholesterol. The results in humans, however, are less clear. This review summarizes the results that have been published on plasma triglyceride, apolipoprotein B, HDL-cholesterol and LDL-cholesterol levels in humans and animal models in response to proanthocyanidin extracts and proanthocyanidin-rich foods. The physiological processes and biochemical pathways that are related to lipid homeostasis and affected by proanthocyanidin consumption are also discussed. Intestinal lipid absorption, chylomicron secretion by the intestine and VLDL secretion by the liver are the processes that are most repressed by proanthocyanidins, which, therefore, induce hypolipidemic effects.

Bioavailability of procyanidin dimers and trimers and matrix food effects in in vitro and in vivo models

Among procyanidins (PC), monomers, such as catechin and epicatechin, have been widely studied, whereas dimer and trimer oligomers have received much less attention, despite their abundance in our diet. Recent studies have showed that as dimers and trimers could be important in determining the biological effects of procyanidin-rich food, understanding their bioavailability and metabolism is fundamental. The purpose of the present work is to study the stability of PC under digestion conditions, the metabolism and the bioavailability by using a combination of in vitro and in vivo models. Simultaneously, the matrix effect of a carbohydrate-rich food on the digestibility and bioavailability of PC is investigated. The results show a high level of stability of PC under gastric and duodenal digestion conditions. However, the pharmacokinetic study revealed limited absorption. Free forms of dimers and trimers have been detected in rat plasma, reaching the maximum concentration 1 h after oral intake of a grape seed extract.

Modulatory effect of grape-seed procyanidins on local and systemic inflammation in diet-induced obesity rats

Chronic low-grade inflammation in obesity is characterized by macrophage accumulation in white adipose tissue (WAT) and abnormal cytokine production. We tested the hypothesis that grape-seed procyanidin extract (PE), with known anti-inflammatory and antioxidant effects, would improve local and systemic inflammation in diet-induced obesity rats. First, we analyzed the preventive effects of procyanidins (30 mg/kg per day) on rats fed a 60% kcal fat diet for 19 weeks. Second, we induced cafeteria diet obesity for 13 weeks to investigate the corrective effects of two PE doses (25 and 50 mg/kg per day) for 10 and 30 days. In the preventive model, PE group had reduced not only body weight but also plasmatic systemic markers of inflammation tumor necrosis factor-α (TNF-α) and C-reactive protein (CRP). The PE preventive treatment significantly showed an increased adiponectin expression and decreased TNF-α, interleukin-6 and CRP expression in mesenteric WAT and muscle TNF-α. A reduced NF-κB activity in liver is also observed which can be related to low expression rates of hepatic inflammatory markers found in PE group. Finally, PE dietary supplementation is linked to a reduced expression of Emr1 (specific marker of macrophage F4/80), which suggests a reduced macrophage infiltration of WAT. In the corrective model, however, only the high dose of PE reduced CRP plasma levels in the short treatment without changes in plasmatic TNF-α. In conclusion, orally ingested PE helps preventing imbalanced obesity cytokine pattern, but its corrective effects need to be further investigated. The dietary regular intake of food or drinks containing procyanidins might help prevent low-grade inflammatory-related diseases.

Metabolomic assessment of the effect of dietary cholesterol in the progressive development of fatty liver disease.

Nonalcoholic fatty liver disease is considered to be the hepatic manifestation of metabolic syndrome and is usually related to high-fat, high-cholesterol diets. With the rationale that the identification and quantification of metabolites in different metabolic pathways may facilitate the discovery of clinically accessible biomarkers, we report the use of (1)H NMR metabolomics for quantitative profiling of liver extracts from LDLr(-/-) mice, a well-documented mouse model of fatty liver disease. A total of 55 metabolites were identified, and multivariate analyses in a diet- and time-comparative strategy were performed. Dietary cholesterol increased the hepatic concentrations of cholesterol, triglycerides, and oleic acid but also decreased the [PUFA/MUFA] ratio as well as the relative amount of long-chain polyunsaturated fatty acids in the liver. This was also accompanied by variations of the hepatic concentration of taurine, glutathione, methionine, and carnitine. Heat-map correlation analyses demonstrated that hepatic inflammation and development of steatosis correlated with cholesterol and triglyceride NMR derived signals, respectively. We conclude that dietary cholesterol is a causal factor in the development of both liver steatosis and hepatic inflammation.

Effects of a grapeseed procyanidin extract (GSPE) on insulin resistance

Flavonoids are beneficial compounds against risk factors for metabolic syndrome, but their effects and the mechanisms on glucose homeostasis modulation are not well defined. In the present study, we first checked the efficacy of grapeseed procyanidin extract (GSPE) for stimulating glucose uptake in insulin-resistant 3T3-L1 adipocytes. Results show that when resistance is induced with chronic insulin treatment, GSPE maintain a higher stimulating capacity than insulin. In contrast, when dexamethasone is used as the resistance-inducing agent, GSPE is less effective. Next we evaluated how effective different GSPE treatments are at improving glucose metabolism in hyperinsulinemic animals (fed a cafeteria diet). GSPE reduced plasma insulin levels. The lower dose (25 mg GSPE/kg body weight per day) administered for 30 days improved the HOmeostasis Model Assessment-insulin resistance index. This was accompanied by down-regulation of Pparg2, Glut4 and Irs1 in mesenteric white adipose tissue. Similarly, a chronic GSPE treatment of insulin-resistant 3T3-L1 adipocytes down-regulated the mRNA levels of those adipocyte markers, although cells were still able to respond to the acute stimulation of glucose uptake. In summary, 25 mg/kg body weight per day of GSPE has a positive long-term effect on glucose homeostasis, and GSPE could be targeted at adipose tissue, where it might directly stimulate glucose uptake. This work also highlights the need to carefully consider the bioactive dose, since a higher dose does not necessarily correlate to a greater positive effect.

Proanthocyanidins Modulate MicroRNA Expression in Human HepG2 Cells

Mi(cro)RNAs are small non-coding RNAs of 18-25 nucleotides in length that modulate gene expression at the post-transcriptional level. These RNAs have been shown to be involved in a several biological processes, human diseases and metabolic disorders. Proanthocyanidins, which are the most abundant polyphenol class in the human diet, have positive health effects on a variety of metabolic disorders such as inflammation, obesity, diabetes and insulin resistance. The present study aimed to evaluate whether proanthocyanidin-rich natural extracts modulate miRNA expression. Using microarray analysis and Q-PCR, we investigated miRNA expression in HepG2 cells treated with proanthocyanidins. Our results showed that when HepG2 cells were treated with grape seed proanthocyanidin extract (GSPE), cocoa proanthocyanidin extract (CPE) or pure epigallocatechin gallate isolated from green tea (EGCG), fifteen, six and five differentially expressed miRNAs, respectively, were identified out of 904 mRNAs. Specifically, miR-30b* was downregulated by the three treatments, and treatment with GSPE or CPE upregulated miR-1224-3p, miR-197 and miR-532-3p. Therefore, these results provide evidence of the capacity of dietary proanthocyanidins to influence microRNA expression, suggesting a new mechanism of action of proanthocyanidins.

Effects of copper exposure upon nitrogen metabolism in tissue cultured Vitis vinifera.

The present study analyses the effects of copper treatment on nitrogen metabolism in a closed system. Sauvignon grapevines were cultured in agar and exposed to copper levels ranging from 0.07 (control) to 5 µg Cu g(-1) medium. Ammonium, nitrate, nitrite, individual and total amino acids and protein contents were determined in root, and leaves. Enzyme activities of nitrate and nitrite reductases, glutamine synthetase, glutamate synthase and glutamate dehydrogenase were also determined. Copper exposure produces a dramatic change in nitrogen metabolism, with a reduction of total nitrogen, which reflects the reduction on nitrate and free amino acid contents in both root and leaves. The assimilation of nitrate the main nitrogen source in the medium, requires nitrate reductase, which is reduced to negligible activity as response to copper exposure. Primary nitrogen metabolism is also reduced in leaves, although to a lesser extent than in roots, which may explain the differences between the two organs in response to copper exposure. An alternative system for assimilation of nitrogen through glutamate dehydrogenase in roots is proposed, while higher levels of ammonium and glutamine may fullfil the needs of organic nitrogen in the leaves.

Changes in lipolysis and hormone-sensitive lipase expression caused by procyanidins in 3T3-L1 adipocytes

OBJECTIVE: To find out whether lipid stores are influenced by phenolic compounds in wine.DESIGN: Differentiated 3T3-L1 cells were treated with catechin, epicatechin or procyanidin extracts with different degrees of polymerization at 150 μM for different periods of time (0.5–24 h).SUBJECTS: Cell line 3T3-L1.MEASUREMENTS: Cellular viability, glycerol-3-phosphate dehydrogenase activity, glycerol release in the medium, HSL mRNA levels, triacylglycerols and protein.RESULTS: Catechin, epicatechin and procyanidin extracts were not toxic for the 3T3-L1 cells in the conditions assayed. Glycerol-3-phosphate dehydrogenase activity was markedly decreased by 150 μM procyanidin extracts. The release of glycerol into the medium was increased in 150 μM procyanidin extract-treated cells and reached a plateau after 15 h exposure. Procyanidins caused a time-dependent reduction in the HSL mRNA levels.CONCLUSIONS: These results suggest that procyanidins from grape and wine affect lipid metabolism whilst their monomers (catechin and epicatechin) do not. This effect is more pronounced when the degree of polymerization is higher. Procyanidin extracts cause a time-dependent reduction in the HSL mRNA levels, inhibit triacylglycerol synthesis and also favour triacylglycerol hydrolysis until the HSL mRNA had reached very low levels.

Lipogenesis is decreased by grape seed proanthocyanidins according to liver proteomics of rats fed a high fat diet

Bioactive proanthocyanidins have been reported to have several beneficial effects on health in relation to metabolic syndrome, type 2 diabetes, and cardiovascular disease. We studied the effect of grape seed proanthocyanidin extract (GSPE) in rats fed a high fat diet (HFD). This is the first study of the effects of flavonoids on the liver proteome of rats suffering from metabolic syndrome. Three groups of rats were fed over a period of 13 weeks either a chow diet (control), an HFD, or a high fat diet supplemented for the last 10 days with GSPE (HFD + GSPE). The liver proteome was fractionated, using a Triton X-114-based two-phase separation, into soluble and membrane protein fractions so that total proteome coverage was considerably improved. The data from isobaric tag for relative and absolute quantitation (iTRAQ)-based nano-LC-MS/MS analysis revealed 90 proteins with a significant (p < 0.05) minimal expression difference of 20% due to metabolic syndrome (HFD versus control) and 75 proteins due to GSPE treatment (HFD + GSPE versus HFD). The same animals have previously been studied (Quesada, H., del Bas, J. M., Pajuelo, D., Díaz, S., Fernandez-Larrea, J., Pinent, M., Arola, L., Salvadó, M. J., and Bladé, C. (2009) Grape seed proanthocyanidins correct dyslipidemia associated with a high-fat diet in rats and repress genes controlling lipogenesis and VLDL assembling in liver. Int. J. Obes. 33, 1007–1012), and GSPE was shown to correct dyslipidemia observed in HFD-fed rats probably through the repression of hepatic lipogenesis. Our data corroborate those findings with an extensive list of proteins describing the induction of hepatic glycogenesis, glycolysis, and fatty acid and triglyceride synthesis in HFD, whereas the opposite pattern was observed to a large extent in GSPE-treated animals. GSPE was shown to have a wider effect than previously thought, and putative targets of GSPE involved in the reversal of the symptoms of metabolic syndrome were revealed. Some of these novel candidate proteins such as GFPT1, CD36, PLAA (phospholipase A2-activating protein), METTL7B, SLC30A1, several G signaling proteins, and the sulfide-metabolizing ETHE1 and SQRDL (sulfide-quinone reductase-like) might be considered as drug targets for the treatment of metabolic syndrome.