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      Ameliorative potential of Ocimum sanctum in chronic constriction injury-induced neuropathic pain in rats

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          Abstract

          The present study was designed to investigate the ameliorative potential of Ocimumsanctum and its saponin rich fraction in chronic constriction injury-induced neuropathic pain in rats. The chronic constriction injury was induced by placing four loose ligatures around the sciatic nerve, proximal to its trifurcation. The mechanical hyperalgesia, cold allodynia, paw heat hyperalgesia and cold tail hyperalgesia were assessed by performing the pinprick, acetone, hot plate and cold tail immersion tests, respectively. Biochemically, the tissue thio-barbituric acid reactive species, super-oxide anion content (markers of oxidative stress) and total calcium levels were measured. Chronic constriction injury was associated with the development of mechanical hyperalgesia, cold allodynia, heat and cold hyperalgesia along with an increase in oxidative stress and calcium levels. However, administration of Ocimumsanctum (100 and 200 mg/kg p.o.) and its saponin rich fraction (100 and 200 mg/kg p.o.) for 14 days significantly attenuated chronic constriction injury-induced neuropathic pain as well as decrease the oxidative stress and calcium levels. It may be concluded that saponin rich fraction of Ocimum sanctum has ameliorative potential in attenuating painful neuropathic state, which may be attributed to a decrease in oxidative stress and calcium levels.

          Translated abstract

          O presente estudo foi delineado para avaliar-se o potencial efeito analgésico de extratos de Ocimum sanctum e uma fração enriquecida em saponinas na dor neuropática induzida pela lesão por constrição crônica de nervos periféricos em ratos. A lesão foi induzida pela colocação de quatro ligaduras apostas ao nervo ciático, proximalmente à sua trifurcação. Hiperalgesia mecânica, alodínia ao frio, hiperalgesia ao calor nas patas e hiperalgesia ao frio na cauda foram determinadas usando-se testes de picada de agulha, exposição à acetona, placa quente e teste de imersão em solução gelada, respectivamente. Análises bioquímicas com conteúdo de espécies reativas ao tiobarburato em tecidos e conteúdo de ânion super-óxido (ambos marcadores de estresse oxidativo) e os níveis totais de cálcio foram realizadas. A lesão por constrição crônica do nervo ciático foi acompanhada de desenvolvimento de hiperalgesia mecânica, alodínia ao frio, hiperalgesia ao calor e ao frio, acompanhadas de aumento de marcadores de estresse oxidativo e níveis de cálcio. A administração de extratos de Ocimum sanctum (100 e 200 mg/kg, via oral) e sua fração enriquecida em saponinas (100 e 200 mg/kg, via oral) por 14 dias atenuou significativamente a dor neuropática associada à constrição crônica do nervo ciático, bem como associou-se à diminuição de estresse oxidativo e níveis de cálcio. Conclui-se que a fração enriquecida em saponinas de Ocimum sanctumtem potencial atenuante da dor neuropática, que poderia ser atribuída à diminuição do estresse oxidativo e níveis teciduais de cálcio.

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          Most cited references78

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          Spared nerve injury: an animal model of persistent peripheral neuropathic pain.

          Peripheral neuropathic pain is produced by multiple etiological factors that initiate a number of diverse mechanisms operating at different sites and at different times and expressed both within, and across different disease states. Unraveling the mechanisms involved requires laboratory animal models that replicate as far as possible, the different pathophysiological changes present in patients. It is unlikely that a single animal model will include the full range of neuropathic pain mechanisms. A feature of several animal models of peripheral neuropathic pain is partial denervation. In the most frequently used models a mixture of intact and injured fibers is created by loose ligation of either the whole (Bennett GJ, Xie YK. A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain 1988;33:87-107) or a tight ligation of a part (Seltzer Z, Dubner R, Shir Y. A novel behavioral model of neuropathic pain disorders produced in rats by partial sciatic nerve injury. Pain 1990;43:205-218) of a large peripheral nerve, or a tight ligation of an entire spinal segmental nerve (Kim SH, Chung JM. An experimental model for peripheral neuropathy produced by segmental spinal nerve ligation in the rat. Pain 1992;50:355-363). We have developed a variant of partial denervation, the spared nerve injury model. This involves a lesion of two of the three terminal branches of the sciatic nerve (tibial and common peroneal nerves) leaving the remaining sural nerve intact. The spared nerve injury model differs from the Chung spinal segmental nerve, the Bennett chronic constriction injury and the Seltzer partial sciatic nerve injury models in that the co-mingling of distal intact axons with degenerating axons is restricted, and it permits behavioral testing of the non-injured skin territories adjacent to the denervated areas. The spared nerve injury model results in early ( 6 months), robust (all animals are responders) behavioral modifications. The mechanical (von Frey and pinprick) sensitivity and thermal (hot and cold) responsiveness is increased in the ipsilateral sural and to a lesser extent saphenous territories, without any change in heat thermal thresholds. Crush injury of the tibial and common peroneal nerves produce similar early changes, which return, however to baseline at 7-9 weeks. The spared nerve injury model may provide, therefore, an additional resource for unraveling the mechanisms responsible for the production of neuropathic pain.
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            A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man.

            A peripheral mononeuropathy was produced in adult rats by placing loosely constrictive ligatures around the common sciatic nerve. The postoperative behavior of these rats indicated that hyperalgesia, allodynia and, possibly, spontaneous pain (or dysesthesia) were produced. Hyperalgesic responses to noxious radiant heat were evident on the second postoperative day and lasted for over 2 months. Hyperalgesic responses to chemogenic pain were also present. The presence of allodynia was inferred from the nocifensive responses evoked by standing on an innocuous, chilled metal floor or by innocuous mechanical stimulation, and by the rats' persistence in holding the hind paw in a guarded position. The presence of spontaneous pain was suggested by a suppression of appetite and by the frequent occurrence of apparently spontaneous nocifensive responses. The affected hind paw was abnormally warm or cool in about one-third of the rats. About one-half of the rats developed grossly overgrown claws on the affected side. Experiments with this animal model may advance our understanding of the neural mechanisms of neuropathic pain disorders in humans.
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              Calcium regulation of neuronal gene expression.

              Plasticity is a remarkable feature of the brain, allowing neuronal structure and function to accommodate to patterns of electrical activity. One component of these long-term changes is the activity-driven induction of new gene expression, which is required for both the long-lasting long-term potentiation of synaptic transmission associated with learning and memory, and the activity dependent survival events that help to shape and wire the brain during development. We have characterized molecular mechanisms by which neuronal membrane depolarization and subsequent calcium influx into the cytoplasm lead to the induction of new gene transcription. We have identified three points within this cascade of events where the specificity of genes induced by different types of stimuli can be regulated. By using the induction of the gene that encodes brain-derived neurotrophic factor (BDNF) as a model, we have found that the ability of a calcium influx to induce transcription of this gene is regulated by the route of calcium entry into the cell, by the pattern of phosphorylation induced on the transcription factor cAMP-response element (CRE) binding protein (CREB), and by the complement of active transcription factors recruited to the BDNF promoter. These results refine and expand the working model of activity-induced gene induction in the brain, and help to explain how different types of neuronal stimuli can activate distinct transcriptional responses.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                aabc
                Anais da Academia Brasileira de Ciências
                An. Acad. Bras. Ciênc.
                Academia Brasileira de Ciências (Rio de Janeiro )
                1678-2690
                March 2015
                : 87
                : 1
                : 417-429
                Affiliations
                [1 ] Punjabi University India
                Article
                S0001-37652015000100417
                10.1590/0001-3765201520130008
                9d1ec0a7-20e1-4f04-a118-bbce7725cd6b

                http://creativecommons.org/licenses/by/4.0/

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                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=0001-3765&lng=en
                Categories
                MULTIDISCIPLINARY SCIENCES

                lesão por constrição crônica,Ocimum sanctum,saponinas,cálcio,estresse oxidativo,chronic constriction injury,Saponin rich fraction,calcium,oxidative stress

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