Characterization of Schinus lentiscifolius Marchand (Anacardiaceae) Bark Extract and Its Effects on Lymphocyte Oxidative Stress and Heat Shock Response.

Schinus lentiscifolius Marchand has been used in folk medicine to treat immunoinflammatory related diseases, which are marked by OS and altered HSR. Our study aimed to evaluate OS and HSR in lymphocytes treated with S. lentiscifolius bark extracts. S. lentiscifolius barks were partitioned with solvents to obtain hexane (SL-HEX), ethyl acetate (SL-ACOET) and methanol (SL-MEOH) extracts, and the presence of bioactive compounds was evaluated by thin layer chromatography. Total phenols were measured by the Folin-Ciocalteu method and flavonoids were identified by HPLC-DAD-ESI-MS/MS. Antioxidant capacity was verified by DPPH method, cell viability by Trypan Blue method, lipid peroxidation by TBARS and HSP70 by immunoblotting. The SL-ACOET extract presented higher content of phenolic compounds and antioxidant activity in vitro. It was able to reduce lipid peroxidation levels in lymphocytes induced by H2 O2 and improved cell viability. The SL-ACOET extract inhibited HSR by a decrease in both intracellular content and release of 70 kDa heat shock proteins (HSP70) and also by decrease extra-to-intracellular HSP70 ratio in lymphocytes submitted to heat shock (2 h, 41 °C). S. lentiscifolius bark extract has antioxidant activity and inhibitory effect on HSR probably due to the presence of polyphenols as the flavonoids quercetin and kaempferol.

Inflammation, oxidative stress and altered heat shock response in type 2 diabetes: the basis for new pharmacological and non-pharmacological interventions.

Type 2 diabetes mellitus (DM2) is a chronic disease characterised by variable degrees of insulin resistance and impaired insulin secretion. Besides, several pieces of evidence have shown that chronic inflammation, oxidative stress, and 70 kDa heat shock proteins (HSP70) are strongly involved in DM2 and its complications, and various pharmacological and non-pharmacological treatment alternatives act in these processes/molecules to modulate them and ameliorate the disease. Besides, uncontrolled hyperglycaemia is related to several complications as diabetic retinopathy, neuropathy and hepatic, renal and cardiac complications. In this review, we address discuss the involvement of different inflammatory and pro-oxidant pathways related to DM2, and we described molecular targets modulated by therapeutics currently available to treat DM2.

Chaperone duality: the role of extracellular and intracellular HSP70 as a biomarker of endothelial dysfunction in the development of atherosclerosis.

The 70-kDa heat shock proteins (HSP70) may provide relevant information about the endothelial dysfunction in cardiovascular diseases. Located in the intracellular milieu (iHSP70), they are essential chaperones that inhibit nuclear factor kappa B activation, stimulate nitric oxide production and superoxide dismutase activity, and inhibit apoptosis. However, under stressful conditions, HSP70 can be released into the extracellular medium (eHSP70) and act as an inflammatory mediator. Although studies have reported the vasoprotective role of iHSP70, the evidence regarding eHSP70 is contradictory. eHSP70 can activate NFκB and activator protein-1, thus stimulating the release of inflammatory cytokines and production of reactive oxygen species. Due to the antagonistic nature of HSP70 according to its location, the eHSP70/iHSP70 ratio (Heck index) has been proposed as a better marker of inflammatory status; however, more studies are required to confirm this hypothesis. Therefore, this review summarises studies that, together, describe the role of HSP70 in endothelial dysfunction.

Ovariectomy reduces the cardiac cytoprotection in rats exposed to particulate air pollutant.

Fine particulate matter (PM2.5) has been considered a risk factor for cardiovascular diseases by inducing an oxidative and inflammatory phenotype. Besides, the reduction of 17ß-estradiol (E2) levels during menopause is a natural risk for cardiovascular outcomes. During the E2 downfall, there is a high requirement of the 70-kDa heat shock proteins (HSP70), which present essential antioxidant, anti-inflammatory, and anti-senescence roles. We investigated if the ovariectomy, an animal model for menopause, could induce additional effects in cardiac health by impairing oxidative and heat shock response parameters of female rats chronically exposed to residual oil fly ash (ROFA; an inorganic fraction of PM2.5). Thus, ROFA was obtained from São Paulo (Brazil) and solubilized it in saline. Further, female Wistar rats were exposed to 50 µL of saline (control group) or ROFA solution (250 µg) (polluted) by intranasal instillation, 5 days/week, 12 weeks. At the 12th week, animals were subdivided into four groups (n = 6 p/group): control, OVX, polluted, and polluted + OVX. Control and polluted were submitted to false surgery, while OVX and polluted + OVX were ovariectomized. ROFA or saline exposure continued for 12 weeks. Ovariectomy reduced the cardiac catalase activity and iHSP70 expression in female rats exposed to ROFA. Neither plasma eHSP72 levels nor H-index (eHSP72 to cardiac iHSP70 ratio) was affected. In conclusion, ovariectomy reduces the cardiac cytoprotection and antioxidant defense, and enhances the susceptibility to premature cellular senescence in rats exposed to ROFA.

Ovariectomy enhances female rats' susceptibility to metabolic, oxidative, and heat shock response effects induced by a high-fat diet and fine particulate matter.

Obesity and exposure to fine particulate matter (air pollutant PM2.5) are important risk factors for metabolic and cardiovascular diseases. They are also related to early menopause. The reduction of 17ß-estradiol (E2) levels during female climacteric, marked by menopause, is of significant concern because of its imminent influence on metabolism, redox and inflammatory status. This complex homeostasis-threatening scenario may induce a heat shock response (HSR) in cells, enhancing the expression of the 70 kDa heat shock protein (HSP70). A failure in this mechanism could predispose women to cardiovascular diseases. In this study, we evaluated if the climacteric could represent an additional risk among obese rats exposed to PM2.5 by worsening lipid, oxidative, and inflammatory parameters and HSP70 in cardiac tissue. We induced obesity in female Wistar rats using a high-fat diet (HFD) (58.3% as fats) and exposed them to 50 µL of saline 0.9% (control, n = 15) or 250 µg residual oil fly ash (ROFA, the inorganic portion of PM2.5) (polluted, n = 15) by intranasal instillation, 5 days/w for 12 weeks. At the 12th week, we subdivided these animals into four groups: control (n = 6), OVX (n = 9), polluted (n = 6) and polluted + OVX (n = 9). OVX and polluted + OVX were submitted to a bilateral ovariectomy (OVX), a surgical model for menopause, while control and polluted received a false surgery (sham). ROFA exposure and HFD consumption were continued for 12 additional weeks, after which the animals were euthanized. ROFA enhanced the susceptibility to ovariectomy-induced dyslipidemia, while ovariectomy predisposed female rats to the ROFA-induced decrease of cardiac iHSP70 expression. Ovariectomy also decreased the IL-6 levels and IL-6/IL-10 in obese animals, reinforcing a metabolic impairment and a failure to respond to unfavorable conditions. Our results support the hypothesis that obese ovariectomized animals are predisposed to a metabolic worsening under polluted conditions and are at higher risk of cardiovascular diseases.

The association of subchronic exposure to low concentration of PM2.5 and high-fat diet potentiates glucose intolerance development, by impairing adipose tissue antioxidant defense and eHSP72 levels.

The subchronic exposure to fine particulate matter (PM2.5) and high-fat diet (HFD) consumption lead to glucose intolerance by different mechanisms involving oxidative stress and inflammation. Under stressful conditions, the cells exert a heat shock response (HSR), by releasing the 72-kDa heat shock proteins (eHSP72), fundamental chaperones. The depletion of the HSR can exacerbate the chronic inflammation. However, there are few studies about the early effects of the association of HFD consumption and exposure to low concentrations of PM2.5 in the oxidative stress and HSR, in the genesis of glucose intolerance. Thus, we divided 23 male B6129SF2/J mice into control (n = 6), polluted (n = 6), HFD (n = 6), and high-fat diet + polluted (HFD + polluted) (n = 5) groups. Control and polluted received a standard diet (11.4% of fats), while HFD and HFD + polluted received HFD (58.3% of fats). Simultaneously, polluted and HFD + polluted received 5 µg/10 µL of PM2.5, daily, 7×/week, while control and HFD were exposed to 10 µL of saline solution 0.9% for 12 weeks. At the 12th week, animals were euthanized. We collected the metabolic tissues to analyze oxidative parameters, total blood to the hematological parameters, and plasma to eHSP72 measurement. The association of HFD and PM2.5 impaired glucose tolerance in the 12th week. Besides, it triggered an antioxidant defense by the adipose tissue, which was negatively correlated with eHSP72 levels. In conclusion, a low concentration of PM2.5 exposure associated with HFD consumption leads to glucose intolerance, by impairing adipose tissue antioxidant defense and systemic eHSP72 levels.

Evaluation of oxidative stress and brain-derived neurotrophic factor levels related to crack-use detoxification.

Crack is a central nervous system stimulant extracted from the Erythroxylum coca plant. It is considered the most potent and addictive form of cocaine, and its euphoric effects are attained within a few seconds after consumption. Alteration of biological markers of oxidative stress and brain-derived neurotrophic factor (BDNF) could be related to the severity of crack withdrawal symptoms in patients undergoing rehabilitation. Thus, the objective of this study was to evaluate if the crack consumption and the drug detoxification process during 14 days in hospitalization regime was able to modify the oxidative status and BDNF levels, in male crack-abstinent patients. The crack detoxification process increased the glutathione (GSH), total thiol content (GST), nitric oxide (NO), and superoxide dismutase (SOD) levels, and reduced the mean BDNF levels. Moreover, a positive correlation was found between the number of hospital admission days and SOD values and between the GST levels and crack-use time after 14 days of detoxification. Furthermore, a negative correlation between the frequency of crack use and NO levels on the first day of hospitalization was also found. In conclusion, the results of this study indicated that crack consumption causes increased oxidative stress in drug users and that the detoxification process during 14 days was sufficient to improve oxidative parameters and antioxidant defenses of the patients, which could positively contribute to rehabilitation process. In addition, we also observed a great variability in the BDNF levels of the patients during the detoxification process, resulting in a reduction in the mean values of this neurotrophin.

Natural Products with Antiplatelet Action.

BACKGROUND: Complex hemostatic mechanisms are involved in the pathophysiology of various diseases, including cardiovascular diseases. Among them, dysregulation of platelet activity is linked to the progression of atherosclerosis and mainly involves platelet aggregation and a decrease in blood flow in the vascular endothelium. The major platelet activation pathways mediated by agonists involve the arachidonic acid pathway, adenosine diphosphate pathway, serotonin pathway, nitric oxide pathway, and action of free radicals on molecules involved in platelet aggregation. These mechanisms have been widely studied and discussed because they are inhibited by the use of medicinal plants in complementary and alternative medicine, thus reducing platelet aggregation. RESULTS: Of the main plants discussed in this review, which have antiplatelet activity, some include saffron, garlic, green tea, St. John's wort, ginger, ginkgo biloba, ginseng, and guavirova. These herbal medicines have phytochemical components, which are directly related to the antiplatelet activity of the plant, such as flavonoids, curcumins, catechins, terpenoids, polyphenols, and saponins. While the majority of the medicinal plants mentioned here were native to the Asian continents, some are distributed worldwide, and found to a smaller extent throughout the American continent, European continent, Mediterranean, African continent, and the Middle East. CONCLUSION: This review showed that several plants and/or compounds exhibit anti-platelet activity, and are therefore potential research targets for developing drugs to treat diseases related to aggregation disorders.

Inhibitory effect of Campomanesia xanthocarpa in platelet aggregation: Comparison and synergism with acetylsalicylic acid.

BACKGROUND AND AIMS: Cardiovascular diseases of thrombotic origin are related to high mortality and standard therapeutic agent used in this case is acetylsalicylic acid (ASA), but serious adverse events may occur. However, recent data has suggested the plant Campomanesia xanthocarpa has antiplatelet activity and could be a viable alternative. In this study we investigated the effects of the encapsulated powder of this plant in human platelet aggregation. METHODS: 23 healthy subjects were randomly divided into three groups: (1) ASA (100mg), (2) C. xanthocarpa (1000mg) or (3) synergism (500mg of C. xanthocarpa plush 50mg of ASA); daily for five days. Antiplatelet activity was determined by turbidimetric method using ADP or arachidonic acid (AA) agonists before, 5 and 8days after treatments. RESULTS: Treatment with C. xanthocarpa and synergism caused a reduction of 8±13.5% and 12.5±5% in platelet aggregation induced by ADP after 5days of treatment, respectively, returning to basal levels after 8days. For AA agonist, 5days of treatment with C. xanthocarpa, ASA or synergism caused a reduction of 46±15%, 36±12% and 69.3±6% in platelet aggregation, respectively, and first two groups returned to baseline values 8days after treatment ended. Synergism group prolonged antiplatelet effect maintaining aggregation reduction after 8days the end of treatment. CONCLUSION: C. xanthocarpa showed antiplatelet action when stimulated by agonist AA, and contributed to the antiplatelet effect when associated with ASA for both agonists, allowing dose reduction to 50mg.

Biomarkers of Subclinical Atherosclerosis and Natural Products as Complementary Alternative Medicine.

Cardiovascular diseases (CVD) are considered the leading cause of morbidity and mortality from chronic diseases in the world. In addition, about 20% of first and recurrent acute myocardial infarctions (MI) are silent. In this context, subclinical atherosclerosis culminates in evident CVD, through the evolution of early risk factors such as hypercholesterolemia, hypertriglyceridemia and others. The main problem in CVD is related to the long-time between the start of the subclinical atherosclerosis and the manifestation of the disease. The identification of subjects at risk of such events is obviously substantial, since identification leads to implementation and compliance with effective preventive measures that reduce such risk. In this sense, this review demonstrates biomarkers as an alternative to early detection of subclinical atherosclerosis. One of the proposed biomarkers is the Ischemia-modified albumin (IMA), being considered a promising biochemical biomarker for atherosclerotic conditions. Another marker that is gaining strength and is associated with the IMA are the advanced oxidation protein products (AOPP), its measurement provides information on the level of exposure to potentially harmful changes to proteins and metabolic control. And last but not least we have nitric oxide as an early marker mainly related to endothelial dysfunction. In this review also is evidenced the use of the Campomanesia xanthocarpa, a plant native to southern region from Brazil extensively used as complementary and alternative medicine, and natural products to reduce protein oxidation and improve the availability of nitric oxide and consequently vascular function, reducing the risk for development of CVD.

Study of oxidative and inflammatory parameters in LDLr-KO mice treated with a hypercholesterolemic diet: Comparison between the use of Campomanesia xanthocarpa and acetylsalicylic acid.

BACKGROUND: Atherosclerosis is an inflammatory disease that affects the arterial wall leading to myocardial, cerebral, and peripheral ischemic syndromes. The use of low doses of aspirin inhibits platelet aggregation and inflammation and prevents cardiovascular mortality. However, ASA may produce hemorrhagic events. Thus, several studies have sought new natural compounds to suppress platelet aggregation without causing serious adverse effects. PURPOSE: In this sense, this study aims to compare the effects of Campomanesia xanthocarpa plant extract with those of acetylsalicylic acid (ASA) on inflammatory parameters observed in homozygous mice knockout for the low-density lipoprotein receptor (LDLr-KO) treated with a hypercholesterolemic diet. MATERIAL AND METHODS: In this study, 28 male LDLr-KO mice were divided into three groups and fed a hypercholesterolemic diet for 4 weeks. Thereafter, the animals that received the hypercholesterolemic diet were treated for 5 days with (1) distilled water, (2) C. xanthocarpa extract, or (3) acetylsalicylic acid. The levels of inflammatory markers were assessed in the blood samples. The gastric tolerability of the animals after oral administration of the treatments was assessed through quantification of the lesions in the gastric mucosa. RESULTS: The levels of proinflammatory cytokines IL-1, IL-6, TNF-α, and INF-γ were reduced to 19.2 ± 3%, 20.4 + 1.3%, 24.7 ± 1.2%, and 20.8 ± 1.7%, respectively, in the group treated with C. xanthocarpa, when compared to control group. Furthermore, treatment with plant extract significantly increased the levels of the anti-inflammatory cytokine IL-10 by 27.3 ± 5.9%, but ASA showed no significant effect on the same cytokines when compared to the control group, with the exception of IL-10, which presented an increase of 8.6 ± 3.5%. Treatments with C. xanthocarpa and ASA also caused significant reductions of 26.4 ± 3% and 38.4± 6% in the serum levels of oxLDL, respectively. However, only treatment with C. xanthocarpa reduced the levels of anti-oxLDL antibodies when compared with the control (25.8 ± 6%). In addition, the analyzed extract did not induce ulcerogenic activity, while ASA induced the formation of lesions. CONCLUSION: In conclusion, treatment with C. xanthocarpa causes anti-inflammatory activity in hypercholesterolemic animals, with results superior to those obtained with the use of ASA.

ANTIOXIDANT ACTIVITY OF BLACKBERRY ( Rubus sp. ) GENOTYPES FROM THE SOUTHERN REGION OF BRAZIL

The antioxidant activity and bioactive compounds of different blackberry fruit genotypes from the major Brazilian producer region (three cultivars and four selections) were evaluated and compared to the Cherokee cultivar. Phenolic and anthocyanic extracts were obtained and evaluated for each fruit genotype. The phenolic extracts of selections 02/96 and 07/001 presented higher antioxidant activity than those of cultivars in most assays. This activity was partially correlated to the higher amount of total phenolics in these samples. Thus, the phenolic compounds are probably the major responsible for the antioxidant activity in the diphenyl-2-picrylhydrazyl radical scavenging assay (DPPH), ferric reducing antioxidant power (FRAP) assay and thiobarbituric acid reactive substances (TBARS) assay. Quercetin seems to be responsible for the antioxidant activity of blackberry phenolic extracts in the ß-carotene bleaching assay. Concerning anthocyanic extracts, the selection 02/96 and Cherokee cultivar from harvest 2007 had higher antioxidant activity than the other genotypes in most assays. Anthocyanins appear to be the major responsible for the antioxidant activity of anthocyanic extracts in the DPPH and FRAP assays, although ascorbic acid also contributed to the DPPH antioxidant activity. Selection 02/96 appears to have higher antioxidant activity than the commercial cultivars cultivated in the southern Brazil and appears to be promising for nutritional and health purposes.

Caracterização físico-química de variedades de amora-preta da região sul do Brasil / Physicochemical characterization of blackberry from the Southern Region of Brazil

A amora-preta (Rubus sp.) é uma fruta cuja exploração comercial está iniciando no Brasil. Seu cultivo iniciou na década de 70 e vem aumentando com a introdução e adaptação de novas cultivares. Porém, pouco se conhece sobre as disparidades geradas na composição e nas características das frutas dessas novas plantas. O objetivo deste trabalho foi avaliar as características físico-químicas de diferentes cultivares ('Tupy', 'Guarani' e 'Cherokee') e seleções (02/96, 07/001 e 03/001) de amora, que estão sendo estudadas para originar cultivares adaptadas à região Sul do Brasil. Foram analisados a cor objetiva, sólidos solúveis (SS), pH, acidez titulável, composição centesimal e ácidos graxos de amoras. As frutas apresentaram umidade entre 84,8 e 90,3%; proteína entre 0,09 e 0,14%, fibra alimentar entre 5,8 e 5,5% e cinzas entre 0,27 e 0,49%. A seleção 02/96 apresentou menor teor de cinzas. Os SS variaram entre 7,3 a 10,2°Brix, a acidez titulável variou entre 1,30 e 1,58% em ácido cítrico e o pH entre 2,8 e 3,1. A seleção 03/001 apresentou menor valor de SS que as demais e menor tendência ao vermelho, mas maior intensidade de cor que a cultivar 'Tupy'. Os ácidos graxos encontrados em maior concentração foram o ácido palmítico (22-29%), oléico (13-32%) e linoléico (15-33%), com diferenças nas concentrações entre os tipos de amora. As variedades de amora-preta avaliadas apresentaram bom valor nutricional com níveis de açúcar e acidez adequados para a industrialização, além de conter ácidos graxos importantes para a manutenção da saúde.

Blackberry (Rubus sp.) is a fruit whose commercial exploitation is starting in Brazil. Its cultivation began in the 70's and is increasing with the introduction and adaptation of new cultivars. However, little is known about the disparities in the composition and characteristics of the fruit from these new plants. The objective of this study was to evaluate the physicochemical characteristics of different cultivars ('Tupy', 'Guarani' and 'Cherokee') and selections (02/96, 07/001 and 03/001) of blackberries, which are being studied to generate cultivars adapted to Southern Region of Brazil. The objective color, soluble solids (SS), pH, titratable acidity, proximate composition and fatty acids of blackberries were evaluated. The fruit humidity ranged between 84.8 and 90.3%; protein between 0.09 and 0.14%, dietary fiber between 5.5 and 5.8% and ash between 0.27 and 0.49%. Selection 02/96 had the lowest ash content. SS ranged from 7.3 to 10.2°Brix, titratable acidity ranged between 1.30 and 1.58% citric acid and pH between 2.8 and 3.1. Selection 03/001 had the lowest SS value and it also had lower redness, but higher color saturation than 'Tupy' cultivar. The fatty acids found at higher concentration were palmitic (22-29%), oleic (13-32%) and linoleic acid (15-33%), with differences in the concentration among blackberry genotypes. The varieties of blackberry evaluated showed good nutritional value with sugar and acidity levels suitable for industrialization. It also contained fatty acids important for maintaining health.