Antinociceptive Effect of Dillenia indica (Linn.) Mediated by Opioid and Cannabinoid Systems: Pharmacological and Chemical Studies.

Dillenia indica (Linn.) has been reported by several biological activities, including anti-inflammatory, antioxidant, antidiabetic, anti-hyperglycemic, antiproliferative, antimutagenic, anticholinesterase, and antimicrobial. In Brazilian traditional medicine, the fruits of D. indica have been used to treat general topical pain and inflammation, but with no scientific validation. Thus, aiming to study its chemical constitution and antinociceptive properties, the crude extract (CE) and fractions obtained from the fruits of D. indica were submitted to an in vivo pharmacological evaluation and a dereplication study by LC-MS/MS analysis, assisted by the Global Natural Product Social Molecular Networking (GNPS). The oral antinociceptive activity of the fruits of D. indica and the possible participation of the opioid and cannabinoid systems were demonstrated in the formalin-induced nociception model. The chemical dereplication study led us to identify several known chemical constituents, including flavonoids, such as caffeoylmalic acid, naringenin, quercetin, and kaempferol. According to literature data, our results are compatible with significant antinociceptive and anti-inflammatory activities. Therefore, the flavonoid constituents of the fruits of D. indica are probably responsible for its antioxidant, anti-inflammatory, and antinociceptive effects mediated by both opioid and cannabinoid systems, confirming its folk use in the treatment and relief of pain.

Litter reduction-induced obesity promotes early depressive-like behavior and elevated prefrontal cortex GFAP expression in male offspring.

AIMS: The present study was developed to investigate how litter reduction-induced obesity promotes early depressive-related behaviors in rodent offspring. MAIN METHODS: We employed a standardized litter size reduction protocol, dividing litters into groups: normal litters (NL), consisting of six males and six females pups and small litters (SL), comprising two males and two females pups. Maternal behavior was monitored during the initial week of lactation. Subsequently, we assessed the pups for weight gain, locomotor activity, social play behavior, and performance in forced swimming test. We further evaluated the weights of retroperitoneal and perigonadal fat tissues, along with the expression of glial fibrillary acidic pprotein (GFAP) in the hippocampus and prefrontal cortex of the offspring. KEY FINDINGS: Our results indicated that litter size reduction led to an increased the maternal behavior. In contrast, offspring from the SL group displayed greater weight gain and increased, retroperitoneal and perigonadal fat. Both male and female rodents in the SL group exhibited decreased social play behavior, and male offspring spent more time immobile during the forced swimming test, suggesting a depressive-like phenotype. Notably, we observed an increase in the GFAP expression in the prefrontal cortex of male rodents, with a trend toward increased expression in the hippocampus. SIGNIFICANCE: Obesity may facilitate the development of early depressive-like behaviors, potentially associated with elevated GFAP expression in the prefrontal cortex.

Design, synthesis, and biological evaluation of new thalidomide-donepezil hybrids as neuroprotective agents targeting cholinesterases and neuroinflammation.

A new series of eight multifunctional thalidomide-donepezil hybrids were synthesized based on the multi-target-directed ligand strategy and evaluated as potential neuroprotective, cholinesterase inhibitors and anti-neuroinflammatory agents against neurodegenerative diseases. A molecular hybridization approach was used for structural design by combining the N-benzylpiperidine pharmacophore of donepezil and the isoindoline-1,3-dione fragment from the thalidomide structure. The most promising compound, PQM-189 (3g), showed good AChE inhibitory activity with an IC50 value of 3.15 µM, which was predicted by docking studies as interacting with the enzyme in the same orientation observed in the AChE-donepezil complex and a similar profile of interaction. Additionally, compound 3g significantly decreased iNOS and IL-1ß levels by 43% and 39%, respectively, after 24 h of incubation with lipopolysaccharide. In vivo data confirmed the ability of 3g to prevent locomotor impairment and changes in feeding behavior elicited by lipopolysaccharide. Moreover, the PAMPA assay evidenced adequate blood-brain barrier and gastrointestinal tract permeabilities with an Fa value of 69.8%. Altogether, these biological data suggest that compound 3g can treat the inflammatory process and oxidative stress resulting from the overexpression of iNOS and therefore the increase in reactive nitrogen species, and regulate the release of pro-inflammatory cytokines such as IL-1ß. In this regard, compound PQM-189 (3g) was revealed to be a promising neuroprotective and anti-neuroinflammatory agent with an innovative thalidomide-donepezil-based hybrid molecular architecture.

Maternal protein malnutrition prolongs sickness behavior in male offspring.

Female rats were fed a normal or hypoproteic diet during the phases of gestation and lactation. The male offspring of these rats were grown to adulthood and used to study the effects of maternal protein malnutrition on progeny. The adult male rats were pretreated with either saline or LPS and subjected to behavioral tests 2 and 6 h after administration. Tumor necrosis factor (TNF-α), corticosterone and body temperature were the parameters used for assessment. Two hours after LPS administration, sickness behavior was developed in all the animals, regardless of maternal protein malnutrition. After 6 h of LPS administration, sickness behavior was more pronounced in the rats that had been subjected to maternal protein malnutrition. Only the rats with maternal protein malnutrition expressed an increase in the plasma levels of TNF-α and corticosterone. Maternal protein malnutrition prolongs sickness behaviors in offspring.

Maternal protein malnutrition induces autism-like symptoms in rat offspring.

Objective: We tested the correlation between maternal protein malnutrition and autistic-like symptoms using behavioral tests in rodents that measure main behavioral characteristics observed in humans with autism spectrum disorder (ASD). Methods: Pregnant female rats were fed a normal diet or a hypoproteic diet during gestation and lactation periods. The litters were weighed every 3 days during lactation, and the offspring were tested in behavioral tasks during infancy (postnatal day (PND) 5: quantification of ultrasonic vocalizations; PND 13: homing behavior test) and adolescence (PND 30-32: open field, hole-board, play social behavior, and object recognition tests) in order to capture the prevalence of some of the core and associated symptoms of ASD. Results: Litters of the hypoproteic diet group had a lesser weight gain during lactation. In addition, pups of dams fed with a hypoproteic diet vocalized less compared to those fed with a normal diet, and they showed impaired social discrimination abilities in the homing behavior test. In adolescence, both male and female offspring of the hypoproteic diet group showed no impairment in locomotor activity; however, they exhibited stereotypic behavior in the hole-board test and a decrease in social play behaviors. Male offspring showed increased interest in exploring a familiar object rather than a novel object. Conclusion: Our results show that maternal protein malnutrition in rats causes offspring behaviors that resemble core and associated ASD symptoms.

Evaluation of anxiolytic-like effect of 7-epiclusianone isolated from Garcinia brasiliensis in mice

ABSTRACT Garcinia brasiliensis Mart., Clusiaceae, species became the target of studies for some years because it has several compounds including polyprenylated benzophenones, as 7-epiclusianone. This benzophenone has several properties, such as leishmanicidal, anti-inflammatory and antinociceptive effects, however still did not be studied anxiolytic activity. For this, the open field and elevated plus maze tests were used in order to evaluate the effect of administration of 7-epiclusianone (isolated from G. brasiliensis) on behavioral performance. Swiss male mice (n = 10 per group) were pre-treated with vegetable oil (10 ml/kg; i.p.) or 7-epiclusianone (1, 3 or 10 mg/kg, i.p.) or diazepam (0.2 mg/kg, i.p.). After 1 h, the animals were submitted to the open field and elevated plus maze tests. The administration of 7-epiclusianone exerted a possible anxiolytic effect in the open field, increased the number of central crossings and anti-tigmotactic effect. In pre-treated group with 7-epiclusianone (10 mg/kg) was also possible to determine a possible anxiolytic effect in the elevated plus maze due to increased permanence of animals in the open arms. The results suggest a possible anxiolytic-like effect presented by the 7-epiclusianone and suggest its potential for the treatment of anxiety.

Cafeteria diet during the gestation period programs developmental and behavioral courses in the offspring.

The objective of this study was to investigate the effect of exposure to maternal consumption of a hyperenergetic, highly palatable diet, known as the cafeteria diet, during the gestation period on the development and behavior of offspring. For this, we used pregnant female mice that were fed a normal or a cafeteria diet during the gestation period. The evaluation of maternal behavior in lactating dams was performed from the second to the eighth day postpartum (PND 2-8). Weight gain, feed intake, and energy intake were recorded during the gestation period. In the offspring, reflex parameters and physical development were evaluated during the lactation period and when they reached adolescence. Behavioral performance was evaluated in light-dark, open-field, and play behavior tests. In addition, biochemical parameters of the dams and the adolescent offspring were evaluated. The cafeteria diet during gestation altered maternal behavior and the onset of physical and neurodevelopmental landmarks and had an impact on emotional and play behavior in adolescent offspring. In conclusion, our results demonstrate that exposure to maternal consumption of a cafeteria diet during the gestation period can program developmental and behavioral courses in the offspring.

Design, synthesis and pharmacological evaluation of N-benzyl-piperidinyl-aryl-acylhydrazone derivatives as donepezil hybrids: Discovery of novel multi-target anti-alzheimer prototype drug candidates.

A new series of sixteen multifunctional N-benzyl-piperidine-aryl-acylhydrazones hybrid derivatives was synthesized and evaluated for multi-target activities related to Alzheimer's disease (AD). The molecular hybridization approach was based on the combination, in a single molecule, of the pharmacophoric N-benzyl-piperidine subunit of donepezil, the substituted hydroxy-piperidine fragment of the AChE inhibitor LASSBio-767, and an acylhydrazone linker, a privileged structure present in a number of synthetic aryl- and aryl-acylhydrazone derivatives with significant AChE and anti-inflammatory activities. Among them, compounds 4c, 4d, 4g and 4j presented the best AChE inhibitory activities, but only compounds 4c and 4g exhibited concurrent anti-inflammatory activity in vitro and in vivo, against amyloid beta oligomer (AßO) induced neuroinflammation. Compound 4c also showed the best in vitro and in vivo neuroprotective effects against AßO-induced neurodegeneration. In addition, compound 4c showed a similar binding mode to donepezil in both acetylated and free forms of AChE enzyme in molecular docking studies and did not show relevant toxic effects on in vitro and in vivo assays, with good predicted ADME parameters in silico. Overall, all these results highlighted compound 4c as a promising and innovative multi-target drug prototype candidate for AD treatment.

Experiencing early life maternal separation increases pain sensitivity in adult offspring.

Maternal separation is a widely accepted model for studying long-term behavioral changes produced by events during early life and its association with changes in pain sensitivity. Thus, our objective was to evaluate sensitivity to pain, under different stimuli in adult male and female rats that had undergone early life maternal separation. Animals were subjected to maternal separation from postnatal day (PND) 2-15. Maternal behavior and litter weight were evaluated during this period. Sensitivity to pain was assessed in offsprings during adulthood by exposing them to stimuli, including formalin (5%; 20µl), a hot plate, and the electronic von Frey test, 4, 7, 10, and 24h after the administration of saline or Freund's complete adjuvant (CFA) injection. Maternal separation did not affect maternal behavior or litter weight during PND 2-15. However, experiencing maternal separation increased pain sensitivity in the rats subjected to formalin by increasing number of flinches and licking time Further, females appeared more sensitive than males to thermal stimuli, as they showed a decrease in latency in the hot plate test. In this test, male and female offsprings that were exposed to early life maternal separation and received the CFA injection also showed a reduction in latency to react to the painful stimuli. In the von Frey test, there was a reduction in withdrawal threshold in maternal separation animals injected with CFA, thus demonstrating a greater sensitivity to the mechanical stimuli. In conclusion, experiencing early life maternal separation increased pain sensitivity in adult offsprings. Thus, our data are important to understand the impact of environmental influences, such as stressful life events during critical developmental periods, on pain vulnerability.

Maternal dipyrone treatment during lactation in mice reduces maternal behavior and increases anxiety-like behavior in offspring.

Dipyrone (metamizole), a powerful drug, is widely used as an analgesic and antipyretic; however, the safety of its use during lactation and the potential impact on the offspring are not well established. This study aimed to determine the effect of maternal dipyrone treatment during lactation on offspring development and emotional behavior and on the dam's maternal behavior. Hence, on postnatal day (PND) 2, drinking water only or drinking water containing dipyrone at doses of 100, 300, and 500mg/kg/day, were offered to lactating mothers up to PND9. Thereafter, all mice were provided regular drinking water. On PND2, all litters were culled to 8 pups (4 males and 4 females). Maternal behavior was evaluated at PND3, 6, 9, and 12, and at PND7 we evaluated locomotor activity in the open field. Reflex parameters and physical development of offspring were evaluated during lactation. At PND7, analysis of ultrasonic vocalization (USV) was performed. When the animals reached adolescence, we evaluated their performance in the open field, elevated plus maze (EPM), and marble burying. Our data demonstrated that maternal dipyrone treatment during lactation not only altered maternal behavior and the onset of physical and neurodevelopmental landmarks but also had an impact on anxiety-like behavior in offspring.

Protein malnutrition during pregnancy alters maternal behavior and anxiety-like behavior in offspring.

OBJECTIVE: The aim of this study was to evaluate the effects of protein malnutrition during pregnancy on maternal behavior, on the early behavior in pups by ultrasonic vocalizations (USVs) emission, and on the behavior of offspring in adulthood in an elevated T-maze. METHODS: Pregnant female rats were fed a normal protein-powdered diet (22% casein; control) or a low-protein (hypoproteic) diet (6% casein; protein restriction) during the first 2 weeks of pregnancy. On the fifth postpartum day (PND5), the number of USV was rated. On PND7, maternal behavior was assessed. Male offspring in adulthood were evaluated for behavioral performance in an elevated T-maze. RESULTS: Our results demonstrated that a hypoproteic diet during early pregnancy increased the maternal behavior, increased the number of USV by pups, and reduced the inhibitory avoidance responses in an elevated T-maze during adulthood. In addition, there was a reduction in weight gain of rats during pregnancy and of offspring during lactation. CONCLUSION: In conclusion, the data found in our study suggest that the increase in USV emitted by pups due to hypoproteic diet during pregnancy accentuated maternal behavior. In addition, an increase in maternal care promoted the reduction in anxiety-like behavior in adult male offspring.

Effect of prenatal ethanol exposure on sexual motivation in adult rats.

Maternal alcohol use during pregnancy adversely affects prenatal and postnatal growth and increases the risk of behavioral deficits. The aim of the present study was to evaluate the effect of prenatal exposure to a moderate dose of alcohol on sexual motivation during adulthood. Rats were prenatally exposed to ethanol by feeding pregnant dams a liquid diet containing 25% ethanol-derived calories on days 6 through 19 of gestation. The controls consisted of pair-fed dams (receiving an isocaloric liquid diet containing 0% ethanol-derived calories) and dams with ad libitum access to a liquid control diet. The sexual motivation of offspring was evaluated during adulthood. The results revealed that the male and female pups of dams treated with alcohol exhibited reduced weight gain, which persisted until adulthood. Both male and female adult animals from dams that were exposed to alcohol showed a reduction in the preference score in the sexual motivation test. Taken together, these results provide evidence of the damaging effects of prenatal alcohol exposure on sexual motivation responses in adulthood.

Continuous central infusion of cannabinoid receptor agonist WIN 55,212-2 decreases maternal care in lactating rats: consequences for fear conditioning in adulthood males.

In lactating rats, maternal behavior consists of several integrated elements designed to promote care and nutrition of the offspring to ensure their survival. Given previous studies showing the role of the brain endocannabinoid system in maternal behavior, the objective of this work was to determine which changes in maternal care occur during continuous intracerebroventricular infusion of a cannabinoid receptor agonist (WIN 55,212-2). Maternal behavior was scored daily in 4 sessions of 72 min each (3 during the light phase and 1 during the dark phase). During drug infusion, the results indicated decreased time spent by the mother licking pups and an increase in feeding time. To determine the extent to which variations in maternal care during the neonatal period affect offspring later in life, we evaluated possible changes in the fear response of offspring once they reached adulthood. Offspring of mothers treated with a cannabinoid agonist presented a lower latency for freezing behavior and increased time spent freezing in a fear test. These results show that treatment with a cannabinoid agonist decreases maternal care, mainly licking, in lactating rats and is associated with increased fear responses in offspring later in life.

The CB1 cannabinoid receptor mediates glucocorticoid-induced effects on behavioural and neuronal responses during lactation.

It has been shown that glucocorticoids can modulate oxytocin (OT) secretion and disrupt maternal behaviour. Because the CB1 receptor (CB1R) has been implicated in some rapid glucocorticoid-induced actions, the present study aimed to evaluate the possible involvement of CB1Rs in maternal behaviour and neuronal activation during lactation. For this purpose, lactating female rats were pre-treated with dexamethasone (DEX) or saline, followed by treatment with AM251, a CB1R antagonist, or vehicle 90 min later. All of the experiments were performed 30 min after the administration of AM251 or vehicle. To evaluate maternal behaviour, the pups were returned to their home cages to the side of the cage opposite the previous nest after 12 h of separation and were filmed for the next 30 min. Aggressive behaviour was evaluated for 10 min following the placement of a male rat in the home cage. For the evaluation of behavioural performance, lactating rats were subjected to a T-maze and open-field tests. The amount of weight gained by the pups was evaluated 15 min after the onset of suckling to determine the amount of milk that they had obtained from the dam. In the central nervous system of lactating rats, c-Fos-positive nuclei were counted in the medial preoptic area, in both the ventral (v) and dorsal (d) parts of the median preoptic nucleus and in the bed nucleus of the stria terminalis (BNST). The number of neurons that were double-labelled for c-Fos/OT was counted in the medial magnocellular subdivision of the paraventricular nucleus, in the periventricular hypothalamic nucleus and in the supraoptic nucleus of the lactating rats. The results show that DEX had the following effects: (1) decreased the amount time the dam spent licking the pups, the amount of time the dam spent in an arched-nursing position and full maternal behaviour; (2) increased the latency to the first attack and decreased front attacks; (3) increased anxiety-like behaviour; and (4) decreased weight gain in the pups. In addition, DEX decreased neuronal activation in all of the investigated hypothalamic and forebrain areas. AM251 administration reversed these parameters, indicating that the behavioural effects and neuronal responses produced by DEX in lactating rats are likely to be mediated by CB1Rs.

Antidepressant-like activity of Sonchus oleraceus in mouse models of immobility tests.

The aim of the present work is to evaluate the putative antidepressant-like effects of hydroethanolic and dichloromethanic extracts from the aerial parts of Sonchus oleraceus (Family Asteraceae) on the performance of male mice in the forced swimming test (FST) and tail suspension test (TST) models predictive of depression. The hydroethanolic and dichloromethanic extracts, both in doses of 30, 100, and 300 mg/kg, were orally administered 1 hour before carrying out the FST or the TST. The immobility time in both the FST and the TST was significantly reduced by acute oral treatment with the extracts (dose range, 100-300 mg/kg), without accompanying changes in ambulation, as assessed in an open-field test. This excluded the possibility that the effect of the extracts is due to an activation of locomotion. The efficacy of the extracts was found to be comparable to that of amitriptyline (10 mg/kg, p.o.). The present study provides evidence for an antidepressant-like effect of the active principle(s) present in the extracts of S. oleraceus in mice. Therefore, a standardized S. oleraceus extract or its purified constituents could be of potential interest for the treatment of depressive disorders.

Evaluation of the antinociceptive activity of extracts of Sonchus oleraceus L. in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Sonchus oleraceus L. has been used to relieve pain in Brazilian folk medicine. AIM OF THE STUDY: Sonchus oleraceus L. has been used to relieve pain in Brazilian folk medicine. This study was conducted to establish the antinociceptive properties of hydroethanolic and dichloromethane extracts from aerial parts of Sonchus oleraceus in mice using chemical and thermal models of nociception. MATERIALS AND METHODS: The formalin, hot plate, and tail immersion tests as well as acetic acid-induced writhing were used to investigate the antinociceptive activity in mice. RESULTS: Given orally, the extracts at test doses of 30-300 mg/kg, produced significant inhibitions on chemical nociception induced by intraperitoneal acetic acid and subplantar formalin since decreased the number of writhing episodes and the time licking. Treatment with the extracts in the same doses produced a significant increase of the reaction time in tail immersion and in the hot plate test. The extracts administered at 300 mg/kg, p.o. had a stronger antinociceptive effect than indomethacin (5mg/kg, p.o.) and morphine (10mg/kg, p.o.). CONCLUSION: The extracts of Sonchus oleraceus markedly demonstrated antinociceptive action in mice, which supports previous claims of its traditional use.

Antinociceptive effect of the extract of Morus nigra leaves in mice.

Morus nigra has been used to relieve pain in Brazilian folk medicine. This study was conducted to establish the antinociceptive properties of dichloromethane extract from leaves of M. nigra. The formalin, hot plate, and tail immersion tests as well as acetic acid-induced writhing were used to investigate the antinociceptive activity in mice. The extract at test doses of 100 and 300 mg/kg, p.o. clearly demonstrated antinociceptive activity in all tests. The extract administered at 300 mg/kg, p.o. had a stronger antinociceptive effect than indomethacin (5 mg/kg, p.o.) and morphine (10 mg/kg, p.o.), which supports previous claims for its traditional use.