Terapia com ozônio no controle de lesões cerebrais em ratos neonatos com hidrocefalia experimental
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Data
2020-07-03
Autores
Rosseto, Laryssa Petrocini [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Resumo- Hidrocefalia é uma doença neurológica comum em neonatos e que está associada à destruição de axônios periventriculares decorrente de processos isquêmicos, oxidativos e inflamatórios consequentes à dilatação ventricular. Nos casos onde a derivação liquórica é inviabilizada, tratamentos conservativos são usados como teraria neuroprotetora. A terapia com ozônio (O3) tem sido associada a efeitos neuroprotetores por controlar processos oxidativos, inflamatórios e degenerativos em lesões isquêmicas ou associadas à idade. O objetivo deste estudo foi investigar, os efeitos da ozonioterapia no comportamento, desenvolvimento motor e astrogliose reativa em encéfalos de ratos jovens com hidrocefalia. Trinta e dois ratos foram separados aleatoriamente em quatro grupos de oito animais sendo: grupo controle (C); grupo controle + O3 (CO3); grupo hidrocefálico não tratado (HNT); grupo hidrocefálico tratado com O3 (HTO3). Os animais dos grupos HNT e HTO3 foram induzidos à hidrocefalia com sete dias de idade por injeção de caulim a 10% na cisterna magna. Com 14 dias de idade, os grupos CO3 e HTO3 receberam quatro tratamentos de O3 medicinal na dose de 0,1mg/kg, na concentração de 5µg/mL via intraperitoneal. Os ratos foram pesados diariamente e expostos a testes de comportamento até o final do experimento. Todos os animais foram submetidos à eutanásia com 28 dias de vida, e seus encéfalos retirados e processados para análise histológica e imunoistoquímica do corpo caloso e matriz germinativa. Ratos do grupo HNT apresentaram pior desempenho em teste de comportamento de campo aberto quando comparados aos ratos C e CO3 (P =0,0202). Além disso, esses animais apresentaram ganho de peso tardio comparado aos demais grupos. Os animais HTO3 obtiveram desempenho semelhante ao grupo C nos testes de campo aberto e maior ganho de peso quando comparado ao grupo HNT. Na avaliação histológica convencional observou-se que a hidrocefalia produziu redução da população de células neurais e sinais sugestivos de morte nas células remanescentes nas estruturas encefálicas estudadas, além de evidências de lesões nas fibras nervosas, com sinais de edema e esgarçamento. Após estudo por imunoistoquímica para GFAP, foi observado também que a hidrocefalia causou reação astrocitária nessas mesmas estruturas. Todas as lesões observadas foram atenuadas após o tratamento com administração de O3. Esses achados indicam que a ozonioterapia possui potencial ação neuroprotetora mediada em parte, pela regulação da reação astrocitária.
Abstract: Hydrocephalus is a common neurological disease in neonates and is associated with the destruction of periventricular axons due to ischemic, oxidative and inflammatory processes resulting from ventricular dilation. In cases where the ventricular shunt is not viable, conservative treatments are used as would be neuroprotective. Ozone therapy (O3) has been associated with neuroprotective effects by controlling oxidative, inflammatory and degenerative processes in ischemic or age-related injuries. The objective of this study was to investigate, the effects of ozone therapy on behavior, motor development and reactive astrogliosis in young rats with hydrocephalus. Thirty-two rats were randomly divided into four groups of eight animals: control group (C); control group + O3 (CO3); untreated hydrocephalic group (HNT); hydrocephalic group treated with O3 (HTO3). The animals of the groups HNT and HTO3 were induced to hydrocephalus at seven days of age by injection of 10% kaolin in the cisterna magna. At 14 days of age, the CO3 and HTO3 groups received four treatments of medicinal O3 at a dose of 0.1mg / kg, at a concentration of 5µg / mL intraperitoneally. The rats were weighed daily and exposed to behavioral tests until the end of the experiment. All animals were euthanized at 28 days of age, and their brains removed and processed for histological and immunohistochemical analysis of the corpus callosum and germinal matrix. Rats from the HNT group showed worse performance in the open field behavior test when compared to C and CO3 rats (P = 0.0202). In addition, these animals showed late weight gain compared to the other groups. The HTO3 animals obtained similar performance to group C in the open field tests and greater weight gain when compared to the HNT group. In the conventional histological evaluation, it was noted that hydrocephalus produced a reduction in the cell population and signs suggestive of death in the remaining cells in the brain structures studied, in addition to evidence of damage to nerve fibers, with signs of edema and fraying. After immunohistochemistry study for GFAP, it was also observed that hydrocephalus caused an astrocytic reaction in these same structures. All lesions observed were alleviated after treatment with O3 administration. These findings indicate that ozone therapy may have a potential neuroprotective action mediated in part by regulation of the astrocytic reaction.
Abstract: Hydrocephalus is a common neurological disease in neonates and is associated with the destruction of periventricular axons due to ischemic, oxidative and inflammatory processes resulting from ventricular dilation. In cases where the ventricular shunt is not viable, conservative treatments are used as would be neuroprotective. Ozone therapy (O3) has been associated with neuroprotective effects by controlling oxidative, inflammatory and degenerative processes in ischemic or age-related injuries. The objective of this study was to investigate, the effects of ozone therapy on behavior, motor development and reactive astrogliosis in young rats with hydrocephalus. Thirty-two rats were randomly divided into four groups of eight animals: control group (C); control group + O3 (CO3); untreated hydrocephalic group (HNT); hydrocephalic group treated with O3 (HTO3). The animals of the groups HNT and HTO3 were induced to hydrocephalus at seven days of age by injection of 10% kaolin in the cisterna magna. At 14 days of age, the CO3 and HTO3 groups received four treatments of medicinal O3 at a dose of 0.1mg / kg, at a concentration of 5µg / mL intraperitoneally. The rats were weighed daily and exposed to behavioral tests until the end of the experiment. All animals were euthanized at 28 days of age, and their brains removed and processed for histological and immunohistochemical analysis of the corpus callosum and germinal matrix. Rats from the HNT group showed worse performance in the open field behavior test when compared to C and CO3 rats (P = 0.0202). In addition, these animals showed late weight gain compared to the other groups. The HTO3 animals obtained similar performance to group C in the open field tests and greater weight gain when compared to the HNT group. In the conventional histological evaluation, it was noted that hydrocephalus produced a reduction in the cell population and signs suggestive of death in the remaining cells in the brain structures studied, in addition to evidence of damage to nerve fibers, with signs of edema and fraying. After immunohistochemistry study for GFAP, it was also observed that hydrocephalus caused an astrocytic reaction in these same structures. All lesions observed were alleviated after treatment with O3 administration. These findings indicate that ozone therapy may have a potential neuroprotective action mediated in part by regulation of the astrocytic reaction.
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Palavras-chave
Estresse oxidativo, Ozônio, Isquemia cerebral, Astrócitos