Evaluation of 12-Week Standardized Beetroot Extract Supplementation in Older Participants: A Preliminary Study of Human Health Safety.

In recent years, there has been a notable surge in the popularity of beetroot-based dietary supplements, driven by their rich nitrate composition. Several types of beetroot-based dietary supplements can be found in markets worldwide; however, ensuring the safety of dietary supplements is a crucial consideration, as there is limited evidence on their safety, especially for older populations. Therefore, the purpose of the current study was to evaluate the safety and tolerability of a nitrate-rich beetroot extract in older participants taking supplements over 12 weeks. The participants were randomly assigned to receive 20 g daily of beetroot extract or a matching placebo. The safety and tolerability of the supplementation were evaluated as the occurrence of adverse events and anthropometric, biochemical, and hemodynamic parameters were measured. No serious adverse events were reported in any group. Anthropometric, biochemical, and hemodynamic parameter changes between the baseline and the end of the study were not statistically significant in either group. However, interestingly, the group receiving beetroot extract supplementation exhibited a notable increase in plasma nitrate levels (p = 0.076, f = 0.50) and showed a decrease in insulin levels (p = 0.026, f = 0.59). In conclusion, we found that 20 g of beetroot extract supplementation for 12 weeks was safe and well tolerated in older participants.

Cytotoxic effects of a triterpene-enriched fraction of Cecropia pachystachya on the human hormone-refractory prostate cancer PC3 cell line.

BACKGROUND: Prostate cancer (PCa) is the most diagnosed invasive cancer and a leading cause of death in men in western countries. Most patients initially respond to androgen deprivation but finally develop hormone-refractory disease, which results in advanced clinical failure and death. Since hormone-refractory disease is highly radiotherapy and chemotherapy resistant, increasing interest has been placed on finding novel therapies for this advanced type of Pca. PURPOSE: The potential cytotoxic effects of the crude extract and fractions obtained from the leaves of Cecropia pachystachya Trécul on different human cancer cell lines were investigated. Additionally, the mechanism of cell death induction of the most active sample (triterpene-enriched fraction, TEF) on the human hormone-refractory prostate PC3 cell line was examined. METHODS: Sulforhodamine B assay was used to measure the viability of human tumor and non-tumor cell lines. To elucidate the mechanism of PC3 cells death induced by TEF, different methodological approaches were used: cell cycle analysis and annexin V/PI staining, nuclear morphological analysis, and senescence-associated-ß-galactosidase assay. Moreover, the mitochondrial membrane potential was measured, and the long-term effects of TEF on PC3 cells were evaluated. RESULTS: TEF exerted cytotoxic effects on PC3 cells but not on human non-tumor cells. The analysis of nuclear morphology of PC3 cells treated with TEF increased the number of cells with large and regular nuclei suggesting senescence induction, which was supported by ß-galactosidase overexpression. Regarding PC3 cells cycle, TEF reduced the number of cells in G1 phase and increased that in sub G0/G1. Apoptosis was not involved in PC3 cell death. However, there was a decrease in mitochondrial membrane potential without the participation of reactive oxygen species (ROS) in the cytotoxic effects detected. Furthermore, there was a decrease in the number of viable cells able to duplicate after long-term TEF treatment. CONCLUSIONS: The results showed the in vitro cytotoxic potential of the triterpene-enriched fraction obtained from the leaves of C. pachystachya on human prostate cancer PC3 cell line.

Lipid-lowering and antiatherogenic effects of Vitex megapotamica (Spreng.) Moldenke in a mice experimental model.

ETHNOPHARMACOLOGICAL RELEVANCE: Vitex megapotamica (Spreng.) Moldenke is a deciduous tree, native of South America. Its leaves are traditionally used to treat cardiovascular diseases. This activity is related to the presence of flavonoids, the major compounds of the crude extract. AIM OF THE STUDY: This study investigated the effects of the oral administration of crude extract and standardized fractions from V. megapotamica leaves on lipid profile and on the formation of atherosclerotic plaque in C57BL/6 LDLr-KO mice treated with high-fat diet (HFD). MATERIALS AND METHODS: Male C57BL/6 LDLr-KO mice were fed with HFD (cholesterol, 1.25%) for 30 days. They were treated with hydroethanolic extract (500 or 1000mg/kg/day) or fractions (125 or 250mg/kg/day). After 30 days of treatment, it was evaluated the serum lipid profile, atherogenic index, and atherosclerotic plaque. RESULTS: All doses of the hydroethanolic extract reduced significantly the levels of total cholesterol, triglycerides, LDL-c and the atherogenic index. The n-butanolic fraction also reduced significantly the levels of total cholesterol, triglycerides, LDL-c and the atherogenic index, at all doses, with exception for the triglycerides, which only the lower dose was effective. The residual fraction reduced significantly the levels of total cholesterol, LDL-c and the atherogenic index, at all doses, with exception for the atherogenic index, which only the higher dose was effective. The atherosclerotic plaque formation was impaired only by the lower dose of the hydroethanolic extract. CONCLUSIONS: Overall, our data suggest that V. megapotamica has potential for the treatment of dyslipidemias.

Development and effect of storage on the stability of enriched flavonoid fraction of Cecropia glaziovii-loaded PLGA nanoparticles.

Studies employing Cecropia glaziovii Snethl leaves have shown great potential in regard to their antiviral activity, mainly related to the phenolic compounds present in this species. The main goal of this work is to combine the therapeutic potential of this species with new technological strategies targeted at the development of an herbal nanoparticulate system for the preparation of a phytotherapeutic formulation. Poly (lactic-co-glycolic acid) nanoparticles containing the enriched flavonoid fraction of Cecropia glaziovii Snethl were developed through a study for the choice of preparation technique, amount of drug and surfactants used. These nanostructured systems were characterized by particle size, polydispersity, zeta potential, encapsulation efficiency and drug-loading capacity. A stability study of the formulations was conducted at room temperature over a period of 60 days. The optimal formulation that best fit the characteristics of the encapsulated material was determined. Sorbitan monooleate and the poloxamer 188 resulted in better colloidal stability, added to the organic and aqueous phases, respectively. These findings suggest that in the field of nanoparticles stability, it is important to evaluate the composition of the nanoparticulate system. This work highlights the importance of the optimization process, searching for a good formulation with suitable structural stabilization.

LC/QTOF profile and preliminary stability studies of an enriched flavonoid fraction of Cecropia pachystachya Trécul leaves with potential antidepressant-like activity.

There is increasing interest in natural antioxidants that are candidates for the prevention of brain damage occurring in major depressive disorders. Cecropia pachystachya is a tropical tree species of Central and South America and a rich source of polyphenols, particularly flavonoids. The aim of this study was to characterize the flavonoid profile of an enriched flavonoid fraction of C. pachystachya (EFF-Cp) and evaluate the antidepressant-like effects of its acute administration in behavior, cytokine levels, oxidative stress and energy metabolism parameters. The EFF-Cp chemical characterization was performed by HPLC/DAD and LC/QTOF. The antidepressant-like effects were performed by the forced swimming test, splash test and open field test. EFF-Cp revealed 15 flavonoids, including seven new glycosyl flavonoids for C. pachystachya. Quantitatively, EFF-Cp showed isoorientin (43.46 mg/g), orientin (23.42 mg/g) and isovitexin (17.45 mg/g) as major C-glycosyl flavonoids. In addition, EFF-Cp at doses 50 and 100 mg/kg reduced the immobility time in the forced swimming test, without changing the locomotor activity and grooming time. In addition, EFF-Cp was able to prevent the oxidative damage in some brain areas. In conclusion, the results of this study suggest that EFF-Cp exerts antidepressant-like effects with its antioxidant properties.

Genotoxic, mutagenic and antigenotoxic effects of Cecropia pachystachya Trécul aqueous extract using in vivo and in vitro assays.

ETHNOPHARMACOLOGICAL RELEVANCE: Cecropia pachystachya is a medicinal plant native to South and Central Americas used to treat asthma and diabetes. AIM OF THE STUDY: In this study, we evaluated the genotoxic, mutagenic and antigenotoxic effects of crude aqueous extract of C. pachystachya (CAE-Cp) leaves. MATERIAL AND METHODS: CAE-Cp was analyzed by the Folin-Ciocalteu method to determine total phenolic and tannin contents. High performance liquid chromatography (HPLC) was used to identify major compounds. Distinct tissues from female and male adult mice were treated with 500-2000mg/kg of CAE-Cp by gavage for the comet assay and micronucleus test analyses. In addition, peripheral blood slides of the group treated with 2000mg/kg CAE-Cp were analyzed 3, 6, and 24h after treatment and were exposed to hydrogen peroxide (ex vivo) to evaluate the genotoxic effect using the comet assay. The Salmonella/microsome assay was carried out against to TA100, TA98, TA97a, TA102, and TA1535 strains in presence and absence of the S9 mix. RESULTS: HPLC showed the presence of chlorogenic acid, isoorientin, orientin, and isovitexin as major compounds. Total phenolic and tannin contents were, respectively, 305.6±0.80 and 144.6±19.04mg of gallic acid equivalent/g of extract. Brain DNA damage was observed in all groups treated with CAE-Cp. The H2O2 challenge indicated genotoxic effect only 6h after the administration of the extract. No increase was detected in micronucleus frequency for any group treated with the extract. Mutagenic effects were detected by Salmonella/microsome assay only in TA102 strain without S9 mix at higher doses. CONCLUSION: The results obtained indicate that CAE-Cp was genotoxic to brain tissue. This result is supported by other papers, showing that compounds present in this extract can cross the blood-brain barrier and act on central nervous system.

Anti-inflammatory and antioxidant activities of aqueous extract of Cecropia glaziovii leaves.

ETHNOPHARMACOLOGICAL RELEVANCE: Cecropia glaziovii Sneth leaves extract is widely used as a traditional folk medicine in Brazil, especially for the treatment of diabetes, and as an antihypertensive and antiinflammatory agent. AIM OF THE STUDY: To investigate the anti-inflammatory and antioxidant properties of crude aqueous extract (CAE) of C. glaziovii leaves. MATERIALS AND METHODS: The in vivo anti-inflammatory and antioxidant effect of the CAE (10-300mg/kg, intragastrically) was investigated in the animal model of pleurisy. The cell migration, proinflammatory cytokines (TNF-α, IL-1ß and IL-6), nitrite/nitrate concentration, myeloperoxidase (MPO) activity, oxidative damage in lipids and proteins, lactate dehydrogenase (LDH) activity and total protein content were also analyzed. Furthermore, the in vitro antioxidant activity of CAE was evaluated by the inhibition of formation of thiobarbituric acid reactive substances (TBARS), induced by free radical generators (H2O2, FeSO4 and AAPH) on a lipid-rich substrate. Hence, the chemical characterizarion of CAE by HPLC was therefore performed. The results showed that the inflammatory process caused by the administration of carragenin (Cg) into the pleural cavity resulted in a substantial increase in inflammatory parameters and oxidative damage. These levels seems to be reversed after CAE treatment in animals with similar results to Dexamethasone (Dex) treatment. Further, the CAE was effective in reducing proinflammatory cytokines, cell infiltrate, MPO activity, nitrite/nitrate concentration, LDH activity, and total protein levels with concomitant attenuation of all parameters associated with oxidative damage induced by Cg. Finally, the CAE presented in vitro antioxidant activity induced by free radical generators at all the concentrations investigated. HPLC analysis confirmed the presence of chlorogenic acid and C-glycosylflavonoids (isoorientin and isovitexin) as the major compounds of the CAE. CONCLUSION: CAE of C. glaziovii exerts significant antiinflammatory and antioxidant activities and this effect can be attributed, at least in part, to the presence of chlorogenic acid and the C-glycosylflavonoids.

Preventive Effect of Cecropia pachystachya Against Ketamine-Induced Manic Behavior and Oxidative Stress in Rats.

Cecropia species are widely used in traditional medicine by its anti-diabetic, anti-hypertensive and anti-inflammatory properties. In the present study, we investigated the neuroprotective and antioxidant effects of the crude aqueous extract from Cecropia pachystachya leaves in a rat model of mania induced by ketamine. The results indicated that ketamine treatment (25 mg/kg i.p., for 8 days) induced hyperlocomotion in the open-field test and oxidative damage in prefrontal cortex and hippocampus, evaluated by increased lipid peroxidation, carbonyl protein formation and decreased total thiol content. Moreover, ketamine treatment reduced the activity of the antioxidant enzymes superoxide dismutase and catalase in hippocampus. Pretreatment of rats with C. pachystachya aqueous extract (200 and 400 mg/kg p.o., for 14 days) or with lithium chloride (45 mg/kg p.o., for 14 days, used as a positive control) prevented both behavioral and pro-oxidant effects of ketamine. These findings suggest that C. pachystachya might be a useful tool for preventive intervention in bipolar disorder, reducing the episode relapse and the oxidative damage associated with the manic phase of this disorder .

Renoprotective Effects of Vitex megapotamica (Spreng.) Moldenke in C57BL/6 LDLr-Null Mice Undergoing High Fat Diet.

Although Vitex megapotamica (Spreng.) Moldenke is used in Brazilian folk medicine as hypolipidemic drug no study has been conducted to evaluate the effects of this species in an experimental model of atherosclerosis. So, the aim of this study was to evaluate the possible renoprotective activity of methanolic extract obtained from Vitex megapotamica (MEVM) using C57BL/6 LDLr-null mice submitted to high fat diet (HFD). MEVM was orally administered at doses of 30, 100, and 300 mg/kg, for three weeks, starting from the 2nd week of HFD. Systolic blood pressure (SBP) and diuretic activity were measured weekly. At the end of experiments the serum lipids, atherogenic index serum (AIS), oxidative stress, and markers of renal function were determined. HFD induced a significant increase in the systolic blood pressure, dyslipidemia, increase in AIS, and lipid peroxidation accompanied by an important reduction in renal function. Treatment with MEVM was able to prevent increase in SBP, total cholesterol, triglycerides, AIS, urea, and creatinine levels in LDLr-null mice. These effects were accompanied by a significant reduction in oxidative stress and renal injury. The data reported here support the potential of Vitex megapotamica as candidate to be an herbal medicine used in cardiovascular or renal diseases.

Antidepressant-like effects of aqueous extract from Cecropia pachystachya leaves in a mouse model of chronic unpredictable stress.

Chronic stressful stimuli influence disease susceptibility to depression, cardiovascular, metabolic and neurodegenerative disorders. The present work investigated antidepressant and antioxidant properties of the aqueous extract from Cecropia pachystachya in a mouse model of chronic unpredictable stress (CUS). Our results indicated that acute administration of the aqueous extract (AE) from C. pachystachya (200 and 400mg/kg, p.o.) produced an antidepressant-like effect in the forced swimming test (FST). The chronic treatment with C. pachystachya extract (200mg/kg, p.o., for 14 days) prevented the depressant-like effect but not the anxiogenic effect induced by CUS. In addition to the behavioral modifications, the 14 days of CUS increased lipid peroxidation in the hippocampus (HP) and prefrontal cortex (PFC), decreased total thiol content and glutathione peroxidase activity in the HP. C. pachystachya AE administration during CUS protocol was able to prevent the oxidative damage induced by stress. However, no changes were observed in the activity of the antioxidant enzymes superoxide dismutase and catalase in the above cited brain areas after the stress protocol and treatment. Our results suggest that C. pachystachya prevented both depressive behavior and oxidative damage induced by CUS, supporting its neuroprotective potential against behavioral and biochemical dysfunctions induced by chronic stress.