Aqueous leaf extract from Luehea divaricata Mart. Modulates oxidative stress markers in the spinal cord of rats with neuropathic pain.

ETHNOPHARMACOLOGICAL RELEVANCE: Reactive oxygen species (ROS) play an important role in neuropathic pain (i.e., pain caused by lesion or disease of the somatosensory system). We showed previously that the aqueous extract prepared from Luehea divaricata leaves, a plant explored by native ethnic groups of Brazil to treat different pathologic conditions, exhibits good antioxidant activity and induces analgesia in rats with neuropathic pain (J Ethnopharmacol, 2020; 256:112761. doi: 10.1016/j.jep.2020.112761). The effect was comparable to that of gabapentin, a drug recommended as first-line treatment for neuropathic pain. However, increasing evidence has indicated the need to accurately determine the oxidative stress level of an individual before prescribing supplemental antioxidants. AIM OF THE STUDY: This study assessed the effects of the oral administration of aqueous extract from leaves of L. divaricata on the sciatic functional index (SFI) and spinal-cord pro-oxidant and antioxidant markers of rats with neuropathic pain. MATERIALS AND METHODS: Placement of four loose chromic thread ligatures around the sciatic nerve produced chronic constriction injury (CCI) of the sciatic nerve, a commonly employed animal model to study neuropathic pain. Aqueous extract from leaves of L. divaricata (100, 300, 500 and 1000 mg/kg), gabapentin (50 mg/kg) and aqueous extract (500 mg/kg) + gabapentin (30 mg/kg) were administrated per gavage daily for 10 or 35 days post-CCI. Antinociception was assessed using the von Frey test while SFI showed functional recovery post-nerve lesion throughout the experimental period. At days 10 and 35 post-surgery, the lumbosacral spinal cord and a segment of the injured sciatic nerve were dissected out and used to determine lipid hydroperoxide levels and total antioxidant capacity (TAC). The spinal cord was also used to determine superoxide anion generation (SAG), hydrogen peroxide (H2O2) levels and total thiol content. RESULTS: As expected, the extract, gabapentin and extract + gabapentin induced antinociception in CCI rats. While no significant functional recovery was found at 10 days post-CCI, a significant recovery was found in SFI of extract-treated CCI rats at 21 and 35 days post-CCI. A significant functional recovery was found already at day 10 post-CCI in gabapentin and gabapentin + extract-treated CCI rats. The extract treatment prevented increases in lipid hydroperoxides levels and TAC in injured sciatic nerve, which were found in this tissue of vehicle-treated rats at 10 days post-CCI. Extract also prevented an increase in SAG, H2O2 and lipid hydroperoxides levels in the spinal cord, which were elevated in this tissue of vehicle-treated rats at 10 and 35 days post-CCI. Extract also prevented a decrease in total thiol content and an increase in TAC in the spinal cord of CCI rats in these same time periods. CONCLUSIONS: Aqueous extract from L. divaricata leaves was demonstrated, for the first time, to improve SFI and modulate oxidative stress markers in injured sciatic nerve and spinal cord of CCI rats. Thus, the antinociceptive effect of the extract involves modulation of oxidative stress markers in injured sciatic nerve and spinal cord.

Aqueous extract from Luehea divaricata Mart. Leaves reduces nociception in rats with neuropathic pain.

ETHNOPHARMACOLOGICAL RELEVANCE: Luehea divaricata, popularly known in Brazil as "açoita-cavalo", has been widely explored by different ethnic groups native to Brazil to treat different pathologic conditions, including inflammatory pain. However, no report could be found on the effect that extract of L. divaricata has on neuropathic pain. This is an important topic because convergent and divergent mechanisms underlie inflammatory vs. neuropathic pain indicate that there may not always be a clear mechanistic delineation between these two conditions. AIM OF THE STUDY: The study aimed to determine antioxidant activity and macronutrient composition of aqueous extract from leaves of L. divaricata, and the effect of oral administration on nociception in rats with chronic constriction injury (CCI) of sciatic nerve-induced neuropathic pain, one of the most commonly employed animal models of neuropathic pain. MATERIALS AND METHODS: The antioxidant activity of the extract was evaluated by total phenolic content and DPPH, ABTS●+ and ORAC methods. Vitexin was determined by HPLC to show that the composition of the extract of the present study is similar to that used in previous studies with this genus. Total sugar and sucrose concentrations were assessed by the anthrone method, while glucose and triacilglycerides were determined using commercially available kits. Fructose concentration was calculated from values for total sugars, glucose and sucrose. Total protein was determined by Bradford assay. The effect on DNA strand breaking was investigated by inhibition of strand breaking of supercoiled DNA by hydroxyl radical. The antinociceptive effects of aqueous extract (100, 300, 500, and 1000 mg/kg, i.g.) were evaluated on thermal and mechanical thresholds for neuropathic pain induced by chronic constriction injury (CCI) of the sciatic nerve in rats. We also compared the antinociceptive effect of the extract (500 mg/kg, i.g.) with that induced by gabapentin (50 mg/kg, i.g.), a first-line clinical treatment for neuropathic pain. The effect of co-administration of extract (500 mg/kg, i.g.) and low-dose gabapentin (30 mg/kg, i.g.) was also assessed. In addition, the effect of the extract on body weight, and blood and hepatic parameters were investigated to reveal possible side effects of treatment. RESULTS: The extract showed high content of total phenol; good reducing capacity for DPPH, ABTS●+ and ORAC assays; presence of vitexin; and a high capacity to inhibit strand breaking of supercoiled DNA. The predominant sugar was sucrose, followed by glucose and fructose. Total protein was greater than triacylglycerides, with the latter being present in a trace amount in the extract. The extract increased the thermal and mechanical thresholds, which was reduced by CCI. The antinociceptive effect was comparable to gabapentin and was also found after co-administration of extract and low-dose gabapentin. No significant change was found in body weight and blood and hepatic indicators after extract treatment. CONCLUSIONS: Aqueous extract from L. divaricata leaves was as effective as gabapentin at attenuating CCI-induced neuropathic pain, indicating for first time the therapeutic potential of this species for this type of pain.

The metabolic and neuroinflammatory changes induced by consuming a cafeteria diet are age-dependent.

OBJECTIVES: To compare the effects of a palatable cafeteria diet on serum parameters and neuroinflammatory markers of young and aged female Wistar rats. METHODS: Three-month-old (young) and 18-month-old (aged) female Wistar rats had access to a cafeteria diet (Caf-Young, Caf-Aged) or a standard chow diet (Std-Young, Std-Aged). RESULTS: The Caf-Young group showed a higher food consumption, weight gain, visceral fat depot, serum insulin and leptin levels, and the insulin resistance index (HOMA-IR) than the Std-Young group. The Caf-Aged group exhibited an increase in interleukin-1 levels in the cerebral cortex and hippocampus. The number of GFAP-positive cells did not differ between the groups, but there was a diet effect in the cerebral cortex and an age effect in the hippocampus. Phospho-tau expression did not differ between the groups. DISCUSSION: The 3- and 18-month-old rats responded differently to a cafeteria diet. Insulin and leptin levels are elevated in young animals fed a cafeteria diet, whereas aged animals are prone to neuroinflammation (indicated by an increase in interleukin-1ß levels). A combination of hypercaloric diet and senescence have detrimental effects on the inflammatory response in the brain, which may predispose to neurological diseases.

Effects of Methanol Fraction from Leaves of Schinus terebinthifolius Raddi on Nociception and Spinal-Cord Oxidative Biomarkers in Rats with Neuropathic Pain.

We determined the antioxidant potential of fractions obtained from leaves of Schinus terebinthifolius, a medicinal plant known in Brazil as aroeira, to select the fraction with the best yield and antioxidant performance. These qualities were found in the methanol fraction (MeF), which was administered intraperitoneally (20 mg/kg/day) for 3 and 10 days to rats with chronic constriction injury (CCI) of the sciatic nerve, a model of neuropathic pain. The MeF increased the mechanical and thermal thresholds that had been lowered by CCI. In parallel, the lumbosacral spinal cord showed an increase in superoxide dismutase but a decrease in glutathione peroxidase and glutathione-S-transferase activities in saline- and MeF-treated CCI rats. Catalase activity decreased only in saline-treated CCI rats for 10 days. Total thiols decreased in saline- and MeF-treated CCI rats. Ascorbic acid increased in these rats at day 3 but only in saline-treated CCI rats at day 10. No change was found in hydrogen peroxide and lipid hydroperoxide. Open-field and elevated plus-maze tests and blood parameters of liver function did not change. Thus, the MeF from leaves of S. terebinthifolius has an antinociceptive action with no toxic effects, and it affects oxidant biomarkers in the spinal cord of rats with CCI.

Methionine Administration in Pregnant Rats Causes Memory Deficit in the Offspring and Alters Ultrastructure in Brain Tissue.

In the present work, we evaluated the effect of gestational hypermethioninemia on locomotor activity, anxiety, memory, and exploratory behavior of rat offspring through the following behavior tests: open field, object recognition, and inhibitory avoidance. Histological analysis was also done in the brain tissue of pups. Wistar female rats received methionine (2.68 µmol/g body weight) by subcutaneous injections during pregnancy. Control rats received saline. Histological analyses were made in brain tissue from 21 and 30 days of age pups. Another group was left to recover until the 30th day of life to perform behavior tests. Results from open field task showed that pups exposed to methionine during intrauterine development spent more time in the center of the arena. In the object recognition memory task, we observed that methionine administration during pregnancy reduced total exploration time of rat offspring during training session. The test session showed that methionine reduced the recognition index. Regarding to inhibitory avoidance task, the decrease in the step-down latency at 1 and 24 h after training demonstrated that maternal hypermethioninemia impaired short-term and long-term memories of rat offspring. Electron microscopy revealed alterations in the ultrastructure of neurons at 21 and 30 days of age. Our findings suggest that the cell morphological changes caused by maternal hypermethioninemia may be, at least partially, associated to the memory deficit of rat offspring.

Epitestosterone- and testosterone-replacement in immature castrated rats changes main testicular developmental characteristics.

Epitestosterone is the 17α-epimer of testosterone and has been described as an anti-androgen, since it inhibits the effects produced by testosterone and dihydrotestosterone via the nuclear androgen receptor (nAR). However, epitestosterone also displays an effect which is similar to the non-classical effect of testosterone, depolarizing the membrane potential of Sertoli cells and inducing a rapid Ca2+ uptake. This study aimed to investigate the effects of a treatment with epitestosterone on developmental parameters of immature rats. Animals were chemically castrated by using the gonadotropin-releasing hormone (GnRH) antagonist cetrorelix and then received a replacement of 7 days with epitestosterone or testosterone. Replacement with either epitestosterone or testosterone restored the anogenital distance (AGD) and testicular weight which had been reduced by chemical castration. The immunocontent of nAR and the nAR-immunoreactivity were reduced by epitestosterone treatment in the testis of both castrated and non-castrated animals. Furthermore, testosterone was unable of changing the membrane potential of Sertoli cells through its non-classical action in the group of animals castrated and replaced with epitestosterone. In conclusion, in relation to the level of protein expression of nAR epitestosterone acts as an anti-androgen. However, it acts in the same way as testosterone when genital development parameters are evaluated. Moreover, in castrated rats epitestosterone suppressed the non-classical response of testosterone, changing the pattern of testosterone signalling via a membrane mechanism in Sertoli cells.

After-effects of consecutive sessions of transcranial direct current stimulation (tDCS) in a rat model of chronic inflammation.

Transcranial direct current stimulation (tDCS) induces cortical excitability changes in animals and humans that can last beyond the duration of stimulation. Preliminary evidence suggests that tDCS may have an analgesic effect; however, the timing of these effects, especially when associated with consecutive sessions of stimulation in a controlled animal experiment setting, has yet to be fully explored. To evaluate the effects of tDCS in inflammatory chronic pain origin immediately and 24 h after the last treatment session, complete Freund's adjuvant (CFA) was injected (100 µl) in the right footpad to induce inflammation. On the 15th day after CFA injection, rats were divided into two groups: tDCS (n = 9) and sham (n = 9). The tDCS was applied for 8 days. The hot plate and Von Frey tests were applied immediately and 24 h after the last tDCS session. Eight 20-min sessions of 500 µA anodal tDCS resulted in antinociceptive effects as assessed by the hot plate test immediately (P = 0.04) and 24 h after the last tDCS session (P = 0.006), for the active tDCS group only. There was increased withdrawal latency in the Von Frey test at 24 h after the last session (P = 0.01). Our findings confirm the hypothesis that tDCS induces significant, long-lasting, neuroplastic effects and expands these findings to a chronic pain model of peripheral inflammation, thus supporting the exploration of this technique in conditions associated with chronic pain and peripheral inflammation, such as osteoarthritis.

Effect of treadmill exercise on serotonin immunoreactivity in medullary raphe nuclei and spinal cord following sciatic nerve transection in rats.

The serotoninergic system modulates nociceptive and locomotor spinal cord circuits. Exercise improves motor function and changes dopaminergic, noradrenergic, and serotonergic central systems. However, the direct relationship between serotonin, peripheral nerve lesion and aerobic treadmill exercise has not been studied. Using immunohistochemistry and optic densitometry, this study showed that the sciatic nerve transection increased the serotoninergic immunoreactivity in neuronal cytoplasm of the magnus raphe nuclei of trained and sedentary rats. In the dorsal raphe nucleus the increase only occurred in sedentary-sham-operated rats. In the spinal cord of trained, transected rats, the ventral horn showed significant changes, while the change in dorsal horn was insignificant. Von Frey's test indicated analgesia in all exercise-trained rats. The sciatic nerve functional index indicated recovery in the trained group. Thus, both the aerobic treadmill exercise training and the nervous lesion appear to contribute to changes in serotonin immunoreactivity.

Sciatic nerve transection increases gluthatione antioxidant system activity and neuronal nitric oxide synthase expression in the spinal cord.

Glutathione (GSH) is a major non-enzymatic antioxidant which is present in all tissues. Its protective actions occur through different pathways such its role as a substrate of antioxidant enzymes, such as glutathione peroxidase (GPx) and glutathione-S-transferase (GST). Nitric oxide (NO) is involved in many physiological processes in the central nervous system, including nociception. In spite of much evidence concerning oxidative and nitrosative stress and neuropathic pain, the exact role of these molecules in pain processing is still unknown. Sciatic nerve transection (SNT) was employed to induce neuropathic pain in rats. Glutathione peroxidase (GPx) and glutathione-S-transferase (GST) activities, glutathione (GSH) content, GSH/GSSG ratio, nitric oxide metabolites (NOx) and neuronal nitric oxide synthase (nNOS) protein expression in the lumbosacral spinal cord were determined. All of these analyses were performed in the SNT and sham groups 1, 3, 7 and 15 days after surgery. There was an increase in GPx activity and in GSH content 3 days after surgery in both sham and SNT groups, but the GSH/GSSG ratio increased only in the SNT group in this time point. nNOS expression was upregulated 7 days post SNT. NOx was detected 1 day after surgery in sham and SNT groups, but at 7 and 15 days, the increase occurred only in SNT animals. These results support the role of the gluthatione system in pain physiology and highlight the involvement of NO as an important molecule related to nociception.