Toxicological and pharmacological effects of Eugenia brasiliensis Lam. (Myrtaceae) leaves in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Eugenia brasiliensis Lam., popularly known as "grumixama" or "Brazilian cherry", is widely used in folk medicine with astringent, diuretic, energizing, anti-rheumatic, and anti-inflammatory properties. AIM OF THE STUDY: Despite its traditional use, detailed toxicological studies of Eugenia brasiliensis are few. Thus, in the current study, we evaluate the toxicological effects of hydroalcoholic extract of Eugenia brasiliensis (HEEb) and its antinociceptive and anti-inflammatory activity. MATERIALS AND METHODS: We used male, and female Swiss mice. Acute toxicity study was performed following the Organization for Economic Cooperation and Development (OECD) guideline 425, and subacute toxicity was assessed following OECD guideline 407. We observed behavioral responses, in addition to hematological, biochemical, and histological evaluations. The antinociceptive and anti-inflammatory activity of HEEb were assessed using the Carrageenan-induced mechanical allodynia and paw edema model. Mechanical allodynia, levels of inflammatory cytokines, and oxidative damage were evaluated. RESULTS: The treatment with HEEb was not able to generate important toxicological alterations. Moreover, doses of 100 and 300 mg/kg of HEEb were able to reduce mechanical allodynia, paw edema, and inflammatory cytokines (TNF-α, IL-1ß, and IL-6), decrease malondialdehyde and increase superoxide dismutase enzyme activity in the paw. CONCLUSIONS: This study demonstrated that HEEb does not present important toxic effects. Additionally, an important antinociceptive, anti-inflammatory, and antioxidant potential were observed.

Cytotoxicity and inflammation induced by Philodryas patagoniensis venom.

The Green racer Philodryas patagoniensis is a snake species from South America and accidents with this genus are often neglected. Therefore, this study aimed to evaluate the toxicological, cytotoxic, and inflammatory potential of P. patagoniensis venom (PpV). The experimental model Artemia salina was used to determine toxicity through the median lethal dose (LD50). Cell viability and genotoxicity were evaluated in human mononuclear cells using the Trypan blue test and the Comet assay, respectively. To assess inflammation, mice had the ventral surface of the right hind paw injected with saline, formalin, and three different concentrations of venom (1, 1.5, and 2 µg. 50 µL-1). LD50 in A. salina was 461 µg. mL-1. PpV caused a significant increase in cell death and genotoxicity in human mononuclear cells at two concentrations (575 and 1150 µg. mL-1). PpV shown also to be a strong agent causing nociception in mice. Paw edema totaled four days at 1.5 µg. 50 µL-1. The hyperalgesia caused by the venom had a long duration in mice, lasting eight days at all concentrations evaluated. Thus, we evaluated for the first time the toxicological potential of PpV in A. salina model and in leukocytes. We concluded that systemic oxidative stress, which we infer to be in the genesis of cytotoxicity and genotoxicity observed in vitro, and the inflammatory process are part of the pathways that trigger the venom damage cascades. Relevant data for both scientific research and clinical medicine. Nonetheless, studies are needed to elucidate these mechanisms.

Traditional uses, phytochemistry, and pharmacology of genus Vitex (Lamiaceae).

Vitex, the genus of the family Lamiaceae, comprises of about 230 species mostly distributed in the warm regions of Europe and temperate regions of Asia. Several Vitex species have been used as folk medicine in different countries for the treatment of various kinds of diseases and ailments. The main aim of this review is to collect and analyze the scientific information available about the Vitex species regarding their chemical constituents and pharmacological activities. The phytochemical investigation of various Vitex species has resulted in the isolation of about 556 chemical constituents belong to various chemical category viz. iridoids, diterpenoids, triterpenoids, flavonoids, lignans, sesquiterpenoids, monoterpenoids, ecdysteroids, and others. The crude extracts of different Vitex species as well as pure phytochemicals exhibited a wide spectrum of in-vitro and in-vivo pharmacological activities. In the present review, the scientific literature data on the ethnopharmacological, phytochemical, and pharmacological investigations on the genus Vitex are summarized. More attention should be given in future research to evaluate the pharmacological potential with detailed mechanism of actions for the pure compounds, extracts of plants from this genus. Moreover, their clinical study is needed to justify their use in modern medicine and to further exploring this genus for new drug discovery.

Dry needling increases antioxidant activity and grip force in a rat model of muscle pain.

BACKGROUND: Muscle pain syndromes (MPS) are one of the main causes of functional, structural and metabolic problems, being associated with tissue oxidative damage. Although dry needling is widely used in the treatment of MPS, there is little scientific evidence of its efficacy and underlying mechanisms of action. OBJECTIVES: To investigate the effects of different dry needling techniques on thermal and mechanical hyperalgesia, locomotor and functional activity, and oxidative stress markers in a rat model of muscle pain. METHODS: A total of 48 male Wistar rats underwent injection of the gastrocnemius muscle with control neutral saline (pH 7) and remained untreated (Saline group), or acidic saline (pH 4) and remained untreated (ASA group) or received pregabalin (PG group), deep needling (DN group), superficial needling (SN group) or twitch needling (TN group) with n = 8 rats per group. Mechanical (von Frey test) and thermal hyperalgesia (acetone test), muscle edema (assessed with a caliper), strength and muscle function (grip force evaluation), surface thermography and locomotor and exploratory activities (open field test) were evaluated. The animals were then euthanized, and the gastrocnemius muscle was excised for assessment of oxidative analyses of lipid peroxidation with thiobarbituric acid reactive species (TBA-RS) and total glutathione (GSH) levels. RESULTS: All treatments significantly improved muscle strength and function when compared to the AS group (p < 0.05). Pregabalin reduced locomotor and exploratory activities, while the TN intervention increased the antioxidant response (p < 0.05). CONCLUSION: Dry needling improved strength, functionality and locomotor activity in a rat model of muscle pain. Twitch needling induced an antioxidant effect.

Sida tuberculata: In vitro cytotoxicity and in vivo anti-inflammatory effect.

ETHNOPHARMACOLOGICAL RELEVANCE: Sida tuberculata R. E. Fries (Malvaceae) is a pioneer species considered a weed in farm fields in Southern Brazil. Widely distributed in South Brazil, S. tuberculata is popularly used to treat inflammatory conditions. AIMS OF THE STUDY: The current study aimed to assess the in vitro cytotoxic and in vivo anti-inflammatory properties of S. tuberculata. MATERIALS AND METHODS: Initially, extracts obtained from leaves (STLE) and roots (STRE) were submitted to cytotoxicity tests using human leukocytes (non-malignant cell line) and HepG2 and MCF-7 (tumor cell lines). In sequence, anti-inflammatory properties were investigated against carrageenan-induced peritonitis model. RESULTS: In vitro analyses displayed a significant decrease in human leukocytes viability without genotoxic damage. IC50 results from tumor cells presented significant decrease in cell viability, slightly more pronounced for STRE. In addition, STLE significantly inhibited the inflammatory and oxidative parameters (TBARS, NPSH, SOD, MPO activity, cell influx, and cytokines release). CONCLUSION: Our findings indicate S. tuberculata extracts have cytotoxic potential more pronounced on tumor cell lines, as well as leaves extract shows a significant reduction in acute inflammation process, as already reported for Sida genus and specifically for this species.

In vivo effects of exposure to Golden trumpet Handroanthus chrysotrichus in mice.

The Golden trumpet Handroanthus chrysotrichus is a tree that presents beneficial health properties against various diseases. Thus, this study aims to verify the toxicity of H. chrysotrichus bark extract, observing the effects of exposure to this extract in mice. For this, mice were separated in groups: saline (sterile solution .9%); H. chrysotrichus crude extract (HCCE) 10; HCCE 50, and HCCE 100 mg. kg-1 (p.o.). We analyzed HCCE effects on acute (single exposure) and subchronic protocol (14 days exposure). After both exposures, acute, and subchronic, we collected samples from blood, brain, liver, and kidney tissues for biochemical evaluation. In addition, after subchronic exposure, we performed behavioral tests. Acute exposure caused an increase of lipid peroxidation in liver tissue. Moreover, we observed a significant carbonyl increase in liver and brain tissues from HCCE 50 mg. kg-1. Kidneys presented carbonyl increase in mice treated with the highest concentration. Besides, creatinine increased in the group of the acute exposure at HCCE 100 mg. kg-1. Total leukocyte count decreased in all concentrations tested. Sub-chronic exposure at HCCE 100 mg. kg-1 caused a decrease in the number of crossing and an increase in its self-grooming frequency in the open field test. In this exposure, the brain and liver had a significant increase in carbonyl levels in all concentrations. We concluded that H. chrysotrichus cause behavioral and biochemical alterations in mice. HCCE primary targets seem to be the liver, kidneys, and white cells.

Evaluation of monosodium iodoacetate dosage to induce knee osteoarthritis: Relation with oxidative stress and pain.

AIM: To determine the dose of monosodium iodoacetate (MIA) required to induce oxidative stress, as well as pain and edema; to confirm the induction of knee osteoarthritis (OA) symptoms in rats by the presence of reactive oxygen species (ROS) and reduction of antioxidant agents; and to verify the presence of histopathological injury in these affected joints. METHOD: Biological markers of oxidative stress, pain, knee edema, and cartilage degeneration provided by different doses of MIA (0.5; 1.0 or 1.5 mg) in rat knee joints were analyzed. The animal evaluations were conducted during 15 days for mechanical and cold hypersensitivity, spontaneous pain and edema. After that, blood serum, intra-articular lavage and structures of knee, spinal cord and brainstem were collected for biochemical analysis; moreover, the knees were removed for histological evaluation. RESULTS: This study demonstrates that the highest dose of MIA (1.5 mg) increased the oxidative stress markers and reduced the antioxidant reactions, both in the focus of the lesion and in distant sites. MIA also induced the inflammatory process, characterized by pain, edema, increase in neutrophil count and articular damage. CONCLUSION: This model provides a basis for the exploration of underlying mechanisms in OA and the identification of mechanisms that may guide therapy and the discovery of OA signals and symptoms.

Predictive antidiabetic activities of plants used by persons with Diabetes mellitus.

BACKGROUND: There are several ways to identify medicinal power of phytoconstituents, such as in silico evaluations. Furthermore, ethnopharmacological researches are important alternatives for the identification of plants with medicinal potential. Significantly, medicinal plants are widely used by persons with Diabetes mellitus (DM) to treat manifestations of this syndrome. OBJECTIVES: i) to investigate the use of medicinal plants for individuals with DM and their health profile; ii) to evaluate in silico possible antidiabetic activities for main phytoconstituents of the commonly used plants. METHODS: A questionnaire was used to measure consumption of medicinal plants. The Prediction of Activity Spectra for Substances (PASS) platform was employed to perform in silico evaluations. In silico predictions for antidiabetic activities were performed with the main compounds identified in a literature review which focused on the more utilized plants. RESULTS: We interviewed 105 persons with DM, most them women (73.34%). Overall mean age was 59.35 years, and 97.14% of them were diagnosed with type 2 DM. An evaluation of the routine exams of the interviewees showed that they have a poor metabolic control. Among the interviewees, 67.62% confirmed the use medicinal plants. Main forms of consumed plant preparation were infusion of leaves and in association with mate (a typical beverage of southern Brazil). Most interviewees consume five or more cups of infusion per day, and when consumed with the mate, 1.73 liters per day. Forty-six medicinal plants were mentioned, and cow's paw (Bauhinia) and jambolan (Syzygium cumini) were the most used. The main informed objective for the plant use was blood glucose control (69.01%). The PASS analysis presented six phytoconstituents with high antidiabetic prediction, especially, vicenin-2, the main phytochemical identified in Passiflora genus (Pa = 0.822). CONCLUSION: Our data show that persons with DM use many plants as a complementary treatment to the traditional medicine. Moreover, part of these plants presented phytoconstituents with antidiabetic potential. These data can serve as a basis for future investigations, with the objective of exploring in vitro and in vivo antidiabetic effects of these plants and its compounds.

In vitro and in silico antioxidant and toxicological activities of Achyrocline satureioides.

ETHNOPHARMACOLOGICAL RELEVANCE: Achyrocline satureioides ("macela or marcela") is a medicinal plant, traditionally collected in "Good Friday" before sunrise. In traditional medicine, dried flowers of A. satureioides are used as anti-dyspeptic, antispasmodic and anti-inflammatory. AIM OF THE STUDY: To evaluate the phytochemical profile and to present an in vitro and in silico approach about toxicity and antioxidant potential of A. satureioides flowers extract and its major phytoconstituents. MATERIALS AND METHODS: Plant were collected according to the popular tradition. Extract were obtained by infusion and analyzed from high-performance liquid chromatography. Toxicity was evaluated in Artemia salina and human lymphocytes. Extract antioxidant activity was determined with total antioxidant capacity, DPPH• and ABTS+• scavenging, ferric reducing antioxidant power, deoxyribose degradation assay, and thiobarbituric acid reactive substances (TBA-RS) assay. TBA-RS inhibitions were evaluated in brain of rats for A. satureioides extract and its major phytoconstituents. Predictions of activity spectra for substances and in silico toxicity evaluation from major phytoconstituents were performed via computer simulation. RESULTS: Chromatographic data indicated isoquercitrin, quercetin and caffeic acid as main compounds in flowers extract. Toxicity tests demonstrated a very low toxic potential of A. satureioides. Extract exhibited antioxidant activities in low concentrations. Both extract and major phytochemicals standards showed protection against lipid peroxidation in brain of rats. Computer simulations pointed some biological activities in agreement with traditional use, as well as some experimental results found in this work. Moreover, in silico toxic predictions showed that the A. satureioides major compounds had low probability for toxic risk. CONCLUSION: Our results indicate that A. satureioides infusion possesses low toxicological potential and an effective antioxidant activity. These findings confirm the traditional use of this plant in the folk medicine.

Effects of Bauhinia forficata Tea on Oxidative Stress and Liver Damage in Diabetic Mice.

This study was designed to evaluate the effects of Bauhinia forficata Link subsp. pruinosa (BF) tea on oxidative stress and liver damage in streptozotocin (STZ)-induced diabetic mice. Diabetic male mice have remained 30 days without any treatment. BF treatment started on day 31 and continued for 21 days as a drinking-water substitute. We evaluated (1) BF chemical composition; (2) glucose levels; (3) liver/body weight ratio and liver transaminases; (4) reactive oxygen species (ROS), lipid peroxidation, and protein carbonylation in liver; (5) superoxide dismutase (SOD) and catalase (CAT) activities in liver; (6) δ-aminolevulinate dehydratase (δ-ALA-D) and nonprotein thiols (NPSH) in liver; (7) Nrf2, NQO-1, and HSP70 levels in liver and pancreas. Phytochemical analyses identified four phenols compounds. Diabetic mice present high levels of NQO-1 in pancreas, increased levels of ROS and lipid peroxidation in liver, and decrease in CAT activity. BF treatment normalized all these parameters. BF did not normalize hyperglycemia, liver/body weight ratio, aspartate aminotransferase, protein carbonyl, NPSH levels, and δ-ALA-D activity. The raised oxidative stress seems to be a potential mechanism involved in liver damage in hyperglycemic conditions. Our results indicated that BF protective effect could be attributed to its antioxidant capacity, more than a hypoglycemic potential.

The influence of Bauhinia forficata Link subsp. pruinosa tea on lipid peroxidation and non-protein SH groups in human erythrocytes exposed to high glucose concentrations.

ETHNOPHARMACOLOGICAL RELEVANCE: Bauhinia forficata (BF) has been traditionally used as tea in folk medicine of Brazil for treatment of Diabetes mellitus (DM). AIM OF THE STUDY: To evaluate the effects of BF leaf tea on markers of oxidative damage and antioxidant levels in an experimental model of hyperglycemia in human erythrocytes in vitro. MATERIALS AND METHODS: Human erythrocytes were incubated with high glucose concentrations or glucose and BF tea for 24h and 48h. After incubation lipid peroxidation and non-protein SH levels were analyzed. Moreover, quantification of polyphenols and flavonoids, iron chelating property, scavenging of DPPH, and prevention of lipid peroxidation in isolated lipids were also assessed. RESULTS: A significant amount of polyphenols and flavonoids was observed. The main components found by LC-MS analysis were quercetin-3-O-(2-rhamnosyl) rutinoside, kaempferol-3-O-(2-rhamnosyl) rutinoside, quercetin-3-O-rutinoside and kaempferol-3-O-rutinoside. BF tea presents important antioxidant and chelating properties. Moreover, BF tea was effective to increase non-protein SH levels and reduce lipid peroxidation induced by high glucose concentrations in human erythrocytes. CONCLUSION: The antioxidant effects of BF tea could be related to the presence of different phenolic and flavonoids components. We believe that these components can be responsible to protect human erythrocytes exposed to high glucose concentrations against oxidative damage.

Cryotherapy reduces skeletal muscle damage after ischemia/reperfusion in rats.

The aim of this study was to analyze the effects of cryotherapy on the biochemical and morphological changes in ischemic and reperfused (I/R) gastrocnemius muscle of rats. Forty male Wistar rats were divided into control and I/R groups, and divided based on whether or not the rats were submitted to cryotherapy. Following the reperfusion period, biochemical and morphological analyses were performed. Following cryotherapy, a reduction in thiobarbituric acid-reactive substances and dichlorofluorescein oxidation levels were observed in I/R muscle. Cryotherapy in I/R muscle also minimized effects such as decreased cellular viability, levels of non-protein thiols and calcium ATPase activity as well as increased catalase activity. Cryotherapy also limited mitochondrial dysfunction and decreased the presence of neutrophils in I/R muscle, an effect that was corroborated by reduced myeloperoxidase activity in I/R muscle treated with cryotherapy. The effects of cryotherapy are associated with a reduction in the intensity of the inflammatory response and also with a decrease in mitochondrial dysfunction.