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.

Effects of vitamin D administration on nociception and spinal cord pro-oxidant and antioxidant markers in a rat model of neuropathic pain

Reactive oxygen species (ROS) are involved in neuropathic pain, a complicated condition after nerve tissue lesion. Vitamin D appears to improve symptoms of pain and exhibits antioxidant properties. We investigated the effects of oral administration of vitamin D3, the active form of vitamin D, on nociception, the sciatic functional index (SFI), and spinal cord pro-oxidant and antioxidant markers in rats with chronic constriction injury (CCI) of the sciatic nerve, a model of neuropathic pain. Vitamin D3 (500 IU/kg per day) attenuated the CCI-induced decrease in mechanical withdrawal threshold and thermal withdrawal latency (indicators of antinociception) and SFI. The vitamin prevented increased lipid hydroperoxide levels in injured sciatic nerve without change to total antioxidant capacity (TAC). Vitamin D3 prevented increased lipid hydroperoxide, superoxide anion generation (SAG), and hydrogen peroxide (H2O2) levels in the spinal cord, which were found in rats without treatment at 7 and 28 days post-CCI. A significant negative correlation was found between mechanical threshold and SAG and between mechanical threshold and H2O2 at day 7. Vitamin D3 also prevented decreased spinal cord total thiols content. There was an increase in TAC in the spinal cord of vitamin-treated CCI rats, compared to CCI rats without treatment only at 28 days. No significant changes were found in body weight and blood parameters of hepatic and renal function. These findings demonstrated, for first time, that vitamin D modulated pro-oxidant and antioxidant markers in the spinal cord. Since antinociception occurred in parallel with oxidative changes in the spinal cord, the oxidative changes may have contributed to vitamin D-induced antinociception.

Age-related changes and effects of regular low-intensity exercise on gait, balance, and oxidative biomarkers in the spinal cord of Wistar rats

The present study aimed to analyze age-related changes to motor coordination, balance, spinal cord oxidative biomarkers in 3-, 6-, 18-, 24-, and 30-month-old rats. The effects of low-intensity exercise on these parameters were also analyzed in 6-, 18-, and 24-month-old rats. Body weight, blood glucose, total cholesterol, and high-density lipoprotein (HDL) cholesterol were assessed for all rats. The soleus muscle weight/body weight ratio was used to estimate skeletal muscle mass loss. Body weight increased until 24 months; only 30-month-old rats exhibited decreased blood glucose and increased total cholesterol and HDL cholesterol. The soleus muscle weight/body weight ratio increased until 18 months, followed by a small decrease in old rats. Exercise did not change any of these parameters. Stride length and step length increased from adult to middle age, but decreased at old age. Stride width increased while the sciatic functional index decreased in old rats. Performance in the balance beam test declined with age. While gait did not change, balance improved after exercise. Aging increased superoxide anion generation, hydrogen peroxide levels, total antioxidant capacity, and superoxide dismutase activity while total thiol decreased and lipid hydroperoxides did not change. Exercise did not significantly change this scenario. Thus, aging increased oxidative stress in the spinal cord, which may be associated with age-induced changes in gait and balance. Regular low-intensity exercise is a good alternative for improving age-induced changes in balance, while beneficial effects on gait and spinal cord oxidative biomarkers cannot be ruled out because of the small number of rats investigated (n=5 or 6/group).

Treatment with ascorbic acid and alpha-tocopherol modulates oxidative-stress markers in the spinal cord of rats with neuropathic pain

Vitamin E (vit. E) and vitamin C (vit. C) are antioxidants that inhibit nociception. The effect of these vitamins on oxidative-stress markers in the spinal cord of rats with chronic constriction injury (CCI) of the sciatic nerve is unknown. This study investigated the effect of intraperitoneal administration of vit. E (15 mg·kg-1·day-1) and vit. C (30 mg·kg-1·day-1), given alone or in combination, on spinal cord oxidative-stress markers in CCI rats. Adult male Wistar rats weighing 200-250 g were divided equally into the following groups: Naive (rats did not undergo surgical manipulation); Sham (rats in which all surgical procedures involved in CCI were used except the ligature), and CCI (rats in which four ligatures were tied loosely around the right common sciatic nerve), which received injections of vitamins or vehicle (saline containing 1% Tween 80) for 3 or 10 days (n=6/each group). The vitamins prevented the reduction in total thiol content and the increase in superoxide-anion generation that were found in vehicle-treated CCI rats. While nitric-oxide metabolites increased in vehicle-treated CCI rats 3 days after surgery, these metabolites did not show significant changes in vitamin-treated CCI rats. In all rats, total antioxidant capacity and hydrogen-peroxide levels did not change significantly. Lipid hydroperoxides increased 25% only in vehicle-treated CCI rats. These changes may contribute to vit. C- and vit. E-induced antinociception, because scavenging reactive oxygen species seems to help normalize the spinal cord oxidative status altered by pain.