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Abelmoschus moschatus extract reverses altered pain and neurohistology of a rat with developmental exposure of fluoride

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Abstract

Fluoride causes major human health complications and evidence suggests that exposure of fluoride during developmental periods adversely affect neurodevelopment, behavior, and maturity of the brain as these periods are critical for developing stages for CNS. The present study aimed to assess the mechanically induced pain sensitivity, cell morphology, circuitry in terms of neural connections and networks and maturation of neurons under fluorideinduced toxicity with concurrent treatment of protective effects of Abelmoschus moschatus seed extract from day 1 of pregnancy to post-natal day (PND) of a rat with age 30th day. Timed pregnant wistar rats (30) were segregated into six groups, viz. control, sodium fluoride (NaF) (20 ppm), NaF + A. moschatus aqueous extract (AMAE), NaF + A. moschatus ethanolic extract (AMEE), AMAE, and AMEE and treated for 51 days (21 gestational and 30 postnatal days). On postnatal days 1, 7, 14, 21 and 30, rats were assessed for oxidative stress markers (GSH and GSSG) and neurohistology of the brain. Increased threshold levels to the mechanical stimulus were observed in fluoridetreated rats. Brain stained with Congo red, Cresyl violet and Golgi cox for β-amyloid, Nissl substance and synaptic connections respectively showed cells become amyloidosis with decreased Nissl substance and decreased number of neuronal connections in NaF exposed rats. Reduced content of GSH and increased GSSG levels (P < 0.001) were also recorded in NaF treated rats. These alterations were associated with increased production of free radicals and the effect of fluoride on the brain is inversely proportional with age. These changes were ameliorated by supplementation with AMAE and AMEE with anti-oxidant properties, which reduce the production of free radicals from fluoride. Thus, the seed extract of A. moschatus had a protective effect over fluoride induced alteration in neural cell maturation, and the establishment of circuitry, mechanical pain sensitivity, and oxidative stress.

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Fluoride is an ineluctable environmental pollutant and its chronic exposure causes nociception and inflammation. Alpha-tocopherol and Selenium (Se) are widely available compounds that are safe if taken in moderation and exert a wide range of antioxidant, analgesic and anti-inflammatory activities. This study examined the protective activity of dietary supplements, alpha-tocopherol (2 mg/kg BW) and Selenium (05 µg/kg BW), by using thermal (Hot plate test, Tail-flick test), chemical (writhing test, formalin test) and neuropathic (allodynia test) tests in fluoride (20mg/kg BW) induced pain models. In addition, anti-inflammatory activity was also assessed with paw oedema assay. The obtained data suggest that hyperalgesia in fluoride exposure group was significantly (p<0.05) exhibited in hot plate, tail flick, writhing response, formalin and allodynia tests. Moreover, inflammation in fluoride exposure group was also significantly (p<0.05) increased in paw oedema tests in comparison with the control group. The combined administration of Se and alpha-tocopherol significantly (p<0.05) increased response latency in hot plate and tail flick tests, reduced writhing responses in the writhing test, increased withdrawal duration in allodynia test, inhibited formalin induced pain response in both phases but it was more pronounced in the second phase and attenuated formalin induced paw oedema in comparison with independent treatment of Se and alpha-tocopherol against NaF suggesting their analgesic and anti-inflammatory activities. These findings conclude the synergistic effects of selenium and alpha-tocopherol against fluoride induced nociception and inflammation.
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Objective: This study reports protective effect of Abelmoschus moschatus seed extract against sodium fluoride-induced neurodegeneration through oxidative stress, neurohistological, and behavioral observations in Wistar rats.Methods: A total of 20 Wistar rats (around 250 g) were randomly classified into four groups, namely, control, fluoride (NaF), fluoride + A. moschatus seed aqueous extract (AMAE), and fluoride + A. moschatus seed ethanol extract (AMEE). The control group animals received normal tap water, fluoride group received fluoridated water at the rate of 40 mg/kg b. wt., 3rd group rats treated with fluoride (40 mg/kg b. wt.) + AMAE (300 mg/kg b. wt.), and 4th group rats treated with fluoride (40 mg/kg b. wt.) + AMEE (300 mg/kg b. wt.). Neurobehavioral responses of rotarod, hot plate, and maze learning tests and oxidantive stress markers including lipid peroxidation (LPO), GSH levels, superoxide dismutase, CAT, and GSH peroxidase (GPx) activities, and also histology with H and E as well as congo red staining were studied in control, fluoride, and A. moschatus seed extract treated against fluoride groups.Results: Decreased neurobehavioral responses with rotarod, hot plate, and maze and enhanced LPO (p<0.05) levels were found in fluoride received animals. Whereas, the superoxide dismutase (SOD), CAT, GSH, and GPx were decreased (p<0.05) in NaF treatment. The rats received seed extract along with NaF showed significant reversal of behavioral and oxidative stress markers and the effect of ethanol extract was more pronounced than aqueous extract. The fluoride-treated group showed disturbed cell structure and reduced number of cells in H and E as well as congo red staining which was reversed in cell morphology and restored cell number in seed extract against NaF-treated group. As a result of increased LPO, decreased antioxidant system, and decreased number of cells, neurodegeneration was observed resulting in the disturbance in functions associated with reported behavior.Conclusion: Okra with high antioxidants activity, seed extract showed reversal of LPO levels and antioxidant status in the brain tissue. And also plant extract administered rats displayed normal cell structure and number of cells than only fluoride received group. Therefore, the aqueous and ethanolic extract of A. moschatus plant seeds has neuroprotective effects against fluoride-induced motor, nociceptive, learning behavior, and on histological structure of brain through antioxidant mechanism. The ethanol extract has shown more efficacy than aqueous extract.
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