Patrícia de Oliveira Carvalho

Phenolic Antioxidants Identified by ESI-MS from Yerba Maté (Ilex paraguariensis) and Green Tea (Camelia sinensis) Extracts

Aqueous extracts of green yerba maté (Ilex paraguariensis) and green tea (Camellia sinensis) are good sources of phenolic antioxidants, as already described in the literature. The subject of this study were organic extracts from yerba maté, both green and roasted, and from green tea. Their phenolic profiles were characterized by direct infusion electrospray insertion mass spectrometry (ESI-MS) and their free radical scavenging activity was determined by the DPPH assay. Organic extracts containing phenolic antioxidants might be used as natural antioxidants by the food industry, replacing the synthetic phenolic additives used nowadays. Ethanolic and aqueous extracts from green yerba maté, roasted yerba maté and green tea showed excellent DPPH scavenging activity (>89%). The ether extracts from green and roasted yerba maté displayed a weak scavenging activity, different from the behavior observed for the green tea ether extract. The main phenolic compounds identified in green yerba maté water and ethanolic extracts were: caffeic acid, quinic acid, caffeoyl glucose, caffeoylquinic acid, feruloylquinic acid, dicaffeoylquinic acid and rutin. After the roasting process two new compounds were formed: caffeoylshikimic acid and dicaffeoylshikimic acid. The ethanolic extracts from yerba maté, both roasted and green, with lower content of phenolic compounds (3.80 and 2.83 mg/mL) presented high antioxidant activity and even at very low phenolic concentrations, ether extract from GT (0.07 mg/mL) inhibited DPPH over 90%.

Antiobesity effects of yerba maté extract (Ilex paraguariensis) in high-fat diet-induced obese mice.

Because the potential of yerba maté (Ilex paraguariensis) has been suggested in the management of obesity, the aim of the present study was to evaluate the effects of yerba maté extract on weight loss, obesity‐related biochemical parameters, and the regulation of adipose tissue gene expression in high‐fat diet–induced obesity in mice. Thirty animals were randomly assigned to three groups. The mice were introduced to standard or high‐fat diets. After 12 weeks on a high‐fat diet, mice were randomly assigned according to the treatment (water or yerba maté extract 1.0 g/kg). After treatment intervention, plasma concentrations of total cholesterol, high‐density lipoprotein cholesterol, triglyceride, low‐density lipoprotein (LDL) cholesterol, and glucose were evaluated. Adipose tissue was examined to determine the mRNA levels of several genes such as tumor necrosis factor‐α (TNF‐α), leptin, interleukin‐6 (IL‐6), C‐C motif chemokine ligand‐2 (CCL2), CCL receptor‐2 (CCR2), angiotensinogen, plasminogen activator inhibitor‐1 (PAI‐1), adiponectin, resistin, peroxisome proliferator‐activated receptor‐γ2 (PPAR‐γ2), uncoupling protein‐1 (UCP1), and PPAR‐γ coactivator‐1α (PGC‐1α). The F4/80 levels were determined by immunoblotting. We found that obese mice treated with yerba maté exhibited marked attenuation of weight gain, adiposity, a decrease in epididymal fat‐pad weight, and restoration of the serum levels of cholesterol, triglycerides, LDL cholesterol, and glucose. The gene and protein expression levels were directly regulated by the high‐fat diet. After treatment with yerba maté extract, we observed a recovery of the expression levels. In conclusion, our data show that yerba maté extract has potent antiobesity activity in vivo. Additionally, we observed that the treatment had a modulatory effect on the expression of several genes related to obesity.

Protective effects of mate tea (Ilex paraguariensis) on H2O2-induced DNA damage and DNA repair in mice

Yerba mate (Ilex paraguariensis) is rich in several bioactive compounds that can act as free radical scavengers. Since oxidative DNA damage is involved in various pathological states such as cancer, the aim of this study was to evaluate the antioxidant activity of mate tea as well as the ability to influence DNA repair in male Swiss mice. Forty animals were randomly assigned to four groups. The animals received three different doses of mate tea aqueous extract, 0.5, 1.0 or 2.0 g/kg, for 60 days. After intervention, the liver, kidney and bladder cells were isolated and the DNA damage induced by H(2)O(2) was investigated by the comet assay. The DNA repair process was also investigated for its potential to protect the cells from damage by the same methodology. The data presented here show that mate tea is not genotoxic in liver, kidney and bladder cells. The regular ingestion of mate tea increased the resistance of DNA to H(2)O(2)-induced DNA strand breaks and improved the DNA repair after H(2)O(2) challenge in liver cells, irrespective of the dose ingested. These results suggest that mate tea could protect against DNA damage and enhance the DNA repair activity. Protection may be afforded by the antioxidant activity of the mate teas bioactive compounds.

Factors that influence the yield and composition of Brazilian propolis extracts

During the extraction of propolis several factors were varied in order to determine how they affected the yield and phenolic composition of the obtained extracts. Six samples of green propolis from the Southeastern region of Brazil were tested. The results indicate that the highest yield of propolis extracts obtained by maceration, which also had the greatest number of components, were those using 70% (v/v) ethanol or more as a solvent. The Soxhlet extraction procedure resulted in even higher yields. No differences were observed between extracts macerated in the presence or absence of light. An increase in yield was observed between extracts macerated for 10 and 30 days although the phenolic content did not vary significantly. The total phenolic content of all extracts varied from 6.41 to 15.24 % but no direct correlation could be found between any of the factors tested.

Effects of Maté Tea (Ilex paraguariensis) Ingestion on mRNA Expression of Antioxidant Enzymes, Lipid Peroxidation, and Total Antioxidant Status in Healthy Young Women

The antioxidant activity of mate tea, the roasted product derived from yerba mate (Ilex paraguarienis), was observed in vitro and in animal models, but studies in humans are lacking. The aim of this study was to investigate the effects of mate tea supplementation on plasma susceptibility to oxidation and on antioxidant enzyme gene expression in healthy nonsmoking women, after acute or prolonged ingestion. We evaluated plasma total antioxidant status (TAS), the kinetics of diene conjugate generation, and thiobarbituric acid reactive substance (TBARS) contents in plasma, as well as mRNA levels of antioxidant gluthatione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT). After the supplementation period with mate tea, lipid peroxidation was acutely lowered, an effect that was maintained after prolonged administration. Total antioxidant status and the level of antioxidant enzyme gene expression were also demonstrated after prolonged consumption. These results suggest that regular consumption of mate tea may increase antioxidant defense of the body by multiple mechanisms.

Maté tea inhibits in vitro pancreatic lipase activity and has hypolipidemic effect on high-fat diet-induced obese mice.

The inhibitory effects of maté tea (MT), a beverage produced with leaves from Ilex paraguariensis, in vitro lipase activity and on obesity in obese mice models were examined. For the in vitro experiment, porcine and human pancreatic lipase (PL) activities were determined by measuring the rate of release of oleic acid from hydrolysis of olive oil emulsified with taurocholate, phospholipids, gum arabic, or polyvinyl alcohol. For the in vivo experiments, animals were fed with a standard diet (SD, n = 10) or high‐fat diet (HFD, n = 30) for 16 weeks. After the first 8 weeks on the HFD, the animals were treated with 1 and 2 g/kg of body weight of MT. The time course of the body weight and obesity‐related biochemical parameters were evaluated. The results showed that MT inhibited both porcine and human PL (half‐maximal inhibitory concentration = 1.5 mg MT/ml) and induced a strong inhibition of the porcine lipase activity in the hydrolysis of substrate emulsified with taurocholate + phosphatidylcholine (PC) (83 ± 3.8%) or PC alone (62 ± 4.3%). MT suppressed the increases in body weight (P < 0.05) and decreased the serum triglycerides and low‐density lipoprotein (LDL)‐cholesterol concentrations at both doses (from 190.3 ± 5.7 to 135.0 ± 8.9 mg/dl, from 189.1 ± 7.3 to 129.3 ± 17.6 mg/dl; P < 0.05, respectively) after they had been increased by the HFD. The liver lipid content was also decreased by the diet containing MT (from 132.6 ± 3.9 to 95.6 ± 6.1 mg/g of tissue; P < 0.05). These results suggest that MT could be a potentially therapeutic alternative in the treatment of obesity caused by a HFD.

Anti-inflammatory effects of yerba mate extract (Ilex paraguariensis) ameliorate insulin resistance in mice with high fat diet-induced obesity

The aim of the present study was to evaluate the effects of yerba maté extract upon markers of insulin resistance and inflammatory markers in mice with high fat diet-induced obesity. The mice were introduced to either standard or high fat diets. After 12 weeks on a high fat diet, mice were randomly assigned to one of the two treatment conditions, water or yerba maté extract at 1.0 gkg(-1). After treatment, glucose blood level and hepatic and soleus muscle insulin response were evaluated. Serum levels of TNF-α and IL-6 were evaluated by ELISA, liver tissue was examined to determine the mRNA levels of TNF-α, IL-6 and iNOS, and the nuclear translocation of NF-κB was determined by an electrophoretic mobility shift assay. Our data show improvements in both the basal glucose blood levels and in the response to insulin administration in the treated animals. The molecular analysis of insulin signalling revealed a restoration of hepatic and muscle insulin substrate receptor (IRS)-1 and AKT phosphorylation. Our data show that the high fat diet caused an up-regulation of the TNF-α, IL-6, and iNOS genes. Although after intervention with yerba maté extract the expression levels of those genes returned to baseline through the NF-κB pathway, these results could also be secondary to the weight loss observed. In conclusion, our results indicate that yerba maté has a potential anti-inflammatory effect. Additionally, these data demonstrate that yerba maté inhibits hepatic and muscle TNF-α and restores hepatic insulin signalling in mice with high fat diet-induced obesity.

Potencial de biocatálise enantiosseletiva de lipases microbianas

Microbial lipases have a great potential for commercial applications due to their stability, selectivity and broad substrate specificity because many non-natural acids, alcohols or amines can be used as the substrate. Three microbial lipases isolated from Brazilian soil samples (Aspergillus niger; Geotrichum candidum; Penicillium solitum) were compared in terms of their stability and as biocatalysts in the enantioselective esterification using racemic substrates in organic medium. The lipase from Aspergillus niger showed the highest activity (18.2 U/mL) and was highly thermostable, retaining 90% and 60% activity at 50 oC and 60 oC after 1 hour, respectively. In organic medium, this lipase provided the best results in terms of enantiomeric excess of the (S)-active acid (ee = 6.1%) and conversion value (c = 20%) in the esterification of (R,S)-ibuprofen with 1-propanol in isooctane. The esterification reaction of the racemic mixture of (R,S)-2-octanol with decanoic acid proceeded with high enantioselectivity when lipase from Aspergillus niger (E = 13.2) and commercial lipase from Candida antarctica (E = 20) were employed.

Enzymatic resolution of (R,S)-ibuprofen and (R,S)-ketoprofen by microbial lipases from native and commercial sources

A enantioseletividade (E) das lipases nativas de Aspergillus niger, Aspergillus terreus, Fusarium oxysporum, Mucor javanicus, Penicillium solitum e Rhizopus javanicus na resolucao dos enantiomeros do (R,S)-ibuprofeno e (R,S)-cetoprofeno na reacao de esterificacao com 1-propanol em isoctano foi comparada com as lipases comerciais de Candida rugosa (Sigma) e Candida antarctica (NovozymO435). A lipase de C. rugosa mostrou boa enantioseletividade (E = 12) comparada com as da NovozymO435 (E = 6.7), de A. niger (E=4.8) e com as outras lipases que foram muito menos seletivas (E por volta de 2.3 e 1.5) na resolucao do (R,S)-ibuprofeno, dentro das condicoes testadas. Apos uma otimizacao preliminar das condicoes da reacao (conteudo de agua, concentracao da enzima e presenca de aditivos) a enantioseletividade da lipase de A. niger pode ser substancialmente aumentada (E = 15). Todas as lipases testadas mostraram baixa seletividade na resolucao de (R,S)-cetoprofeno, resultando baixos rendimentos de ester e de excesso enantiomerico do acido nao esterificado.

Green Tea Extract Supplementation Induces the Lipolytic Pathway, Attenuates Obesity, and Reduces Low-Grade Inflammation in Mice Fed a High-Fat Diet

The aim of this study was to evaluate the effects of green tea Camellia sinensis extract on proinflammatory molecules and lipolytic protein levels in adipose tissue of diet-induced obese mice. Animals were randomized into four groups: CW (chow diet and water); CG (chow diet and water + green tea extract); HW (high-fat diet and water); HG (high-fat diet and water + green tea extract). The mice were fed ad libitum with chow or high-fat diet and concomitantly supplemented (oral gavage) with 400 mg/kg body weight/day of green tea extract (CG and HG, resp.). The treatments were performed for eight weeks. UPLC showed that in 10 mg/mL green tea extract, there were 15 μg/mg epigallocatechin, 95 μg/mg epigallocatechin gallate, 20.8 μg/mg epicatechin gallate, and 4.9 μg/mg gallocatechin gallate. Green tea administered concomitantly with a high-fat diet increased HSL, ABHD5, and perilipin in mesenteric adipose tissue, and this was associated with reduced body weight and adipose tissue gain. Further, we observed that green tea supplementation reduced inflammatory cytokine TNFα levels, as well as TLR4, MYD88, and TRAF6 proinflammatory signalling. Our results show that green tea increases the lipolytic pathway and reduces adipose tissue, and this may explain the attenuation of low-grade inflammation in obese mice.