©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 18 (4): gmr18213 M.L.A. Cirino et al. 2 and miR-222. We did not observe significant differences in miRNA expression in brain tissue in rats submitted to cerebral ischemia, physical exercise and both treatments when compared with the control group. However, miR-222 expression increased in the cerebral ischemia group submitted to physical exercise, which may help promote cerebrovascular regeneration.
Cerebral ischemia has not only a high mortality rate, which is the second leading cause of death worldwide, but is also responsible for severe disabilities in working age individuals, generating enormous public expending for treatment and rehabilitation of the affected individuals. The role of microRNAs in the pathophysiology of cerebral ischemia has been highlighted in current investigations. In addition, recent studies have also highlighted physical exercise as a possible protective factor both in the prevention and in the effects of cerebral ischemia, placing it as an important study resource. Thus, we investigated the role of physical exercise in experimental cerebral ischemia associated with the expression of microRNA-27b. 16 animals were used, divided into four experimental groups: Control, Physical Exercise, Cerebral Ischemia and Cerebral Ischemia associated with Physical Exercise. The real-time PCR methodology was used to analyze the expression of microRNA-27b. Although there were no statistically significant differences in the expression of microRNA-27b between the groups studied, the increased expression of microRNA-27b in the Physical Exercise group indicates its neuroprotective role in the pathophysiology of cerebral ischemia.
Stroke is a major public health problem, with high mortality rates, and a high frequency of disability. Between 1990 and 2010, stroke increased from the fifth to the third leading cause of disability. In addition, its incidence has increased among younger people, with severe health consequences and increased social costs. There is evidence that physical exercise promotes neuroprotective effects when used as a therapeutic treatment. However, the mechanisms of neuroprotection are not yet well known. The objective of the study was to evaluate the expression profile of microRNAs miR-16, miR-21 and miR-155 and the CASPASE-3 and Bcl-2 genes previously related to apoptosis in the tissue (ischemic focus). Forty-eight Wistar rats were divided into four experimental groups: control group, ischemia group, ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 19 (4): gmr18594 L.B. Porsani et al. 2 exercise group and exercise + ischemia group. Before the ischemic procedure, the animals in the exercise and exercise + ischemia groups were submitted to a treadmill training protocol for four weeks. The training lasted 30 min a day at a speed of 18 m / min. For real-time PCR analysis, a fragment of the ischemic area was collected from each animal using a punch to analyze the expression of miRNAs; miR16, miR-21, miR-155, CASPASE-3 and Bcl-2 genes. In the animals that had physical exercise, there appeared to be a neuroprotective effect by the action of microRNAs and CASPASE-3, although no significant difference was observed. Further studies are needed to elucidate the role of apoptosis mechanisms in cerebral ischemia associated with physical exercise, as well as the role of microRNAs in the modulation of targets associated with this mechanism.
Introdução: O acidente vascular cerebral (AVC) é um grande problema de saúde pública, com altas taxas de mortalidade e altos níveis de incapacidade. Entre 1990 e 2010, o AVC aumentou da quinta para a terceira principal causa de incapacidade. Além disso, sua incidência tem aumentado entre pessoas mais jovens, situação em que existem sequelas significativas na saúde e aumento nos custos sociais. Há evidências de que o exercício físico promove efeitos neuroprotetores quando utilizado como tratamento terapêutico em modelo de isquemia experimental. Entretanto, os mecanismos de neuroproteção ainda não estão bem elucidados Objetivo: O objetivo deste estudo foi avaliar por PCR em tempo real o perfil de expressão dos microRNAs miR-21, miR-16, miR-155 no sangue e no tecido (foco isquêmico) em ratos, após a realização dos treinamentos físicos pré-isquemia cerebral focal. Materiais e Métodos: Foram utilizados 48 ratos da linhagem Wistar divididos em quatro grupos experimentais: grupo controle, grupo isquemia, grupo exercício e grupo exercício + isquemia. A metodologia de PCR em tempo real foi utilizada para analisar a expressão dos miRNAs; miR16, miR-21, miR-155, CASPASE-3 e Bcl-2. Resultados e conclusões: Em nosso estudo não observamos diferenças estatísticas significativas na expressão dos miRNAs: miR-16, miR-21, miR-155 e dos genes BCL-2 e CASPASE-3 no tecido cerebral nos grupos submetidos à isquemia cerebral e ao exercício físico quando comparados ao grupo controle.
Stroke is one of the main causes of death and disability worldwide. The great impact on the quality of life of the population and on the health system justifies that we seek relevant alternatives to reduce the incidence and improve the treatment and recovery of patients affected by this disease. Physical exercise appears as an important tool in this scenario, being already pointed out as a possible therapeutic approach for the prevention of non-contagious chronic diseases. In this context, biomarkers such as miRNAs that respond to physical exercise and are directly related to several epigenetic mechanisms appear. Therefore, explaining the molecular mechanisms involved during physical exercise will lead to a better understanding of each stimulus and the dose to be used to better respond to each situation, thus being a promising approach for the evolution of prescription and control of training and processes recovery from various diseases, including stroke. Forty-eight Wistar rats were used, divided into four experimental groups: control group, ischemia group, physical exercise group and exercise + ischemia group. Real-time PCR methodology was used to analyze the expression of miRNAs: miR-126, miR-133b and miR-221. In our study we observed a significant difference in the expression of miR-221 between the control group and the others groups. However, microRNAs: miR-126 and miR-133b do not show significant differences in expression between groups.
CIRINO, M. L. A.; CARVALHO, C. A. M.; NOVAIS, P. C.; FIGUEREDO, R. Z.; PORSANI, L. B.; GULA, I. S.; RODRIGUES, A. R.; SILVA, J. P.; PEREIRA, A. L.; NETO, F. S. L.; SCHIMMING, B. C.; TAZIMA, M. F. G. S.; FAZAN, V. P. S.; CARLOTTI, JR., C. G.; TIRAPELLI, L. F. & COLLI, B. O. Morphometric analysis and pattern of protein and gene expression of apoptosis in focal cerebral ischemia in rats and the neuroprotetive action of hypothermia and ketoprofen. Int. J. Morphol., 38(3):523-529, 2020. SUMMARY:This study aimed to investigate the morphometric and the pattern of protein and gene expression related to the extrinsic apoptotic pathway in experimental focal cerebral ischemia and the hole of neuroprotection with hypothermia and ketoprofen. For this analysis, 120 rats were randomly divided into 3 groups (20 animals each): control -no surgery (20 animals); sham -simulation of surgery (20 animals); ischemic -focal ischemia for 1 hour, without reperfusion (80 animals) and divided into four subgroups with 20 animals each: ischemic + intraischemic hypothermia; ischemic + previous intravenous ketoprofen, and ischemic + hypothermia and ketoprofen. The infarct volume was measured using morphometric analysis of infarct areas defined by triphenyl tetrazolium chloride and the patterns of expression of the apoptosis genes (Fas, c-Flip, caspase-8 and caspase-3) and the apoptosis protein caspase-3 were evaluated by quantitative real-time PCR and immunohistochemistry, respectively. Hypo expression of genes of extrinsic pathway of apoptosis was observed: Fas receptor, c-Flip and caspase-8 in the ischemics areas. Increases in the gene and protein caspase-3 in the ischemic areas were also observed, and these increases were reduced by hypothermia and ketoprofen, also noted in the morphometric study. The caspases-3 increase suggests that this gene plays an important role in apoptosis, probably culminating in cell death and that the neuroprotective effect of hypothermia and ketoprofen is involved. TIRAPELLI, D. P. C.; CIRINO, M. L. A.; CARVALHO, C. A. M.; NOVAIS, P. C.; FIGUEREDO, R. Z.; PORSANI, L. B.; GULA, I. S.; RODRIGUES, A. R.; SILVA, J. P.; PEREIRA, A. L.; NETO, F. S. L.; SCHIMMING, B. C.; TAZIMA, M. F. G. S.; FAZAN, V. P. S.; CARLOTTI, JR., C. G.; TIRAPELLI, L. F. & COLLI, B. O. Morphometric analysis and pattern of protein and gene expression of apoptosis in focal cerebral ischemia in rats and the neuroprotetive action of hypothermia and ketoprofen. Int. J. Morphol., 38(3):523-529, 2020.
P.C. Novais et al. 2 (IR), Alcoholic (A); Alcoholic and ischemic (I + A); Ischemiareperfusion and Alcoholic (IR+A). The blood samples were collected for gene expression analysis of some serum miRNAs by PCR in real time. The serum expression of miRNA-16 was higher in the IR group compared to C and S groups (P < 0.05) but no association with chronic alcoholism was found. The serum expressions of miRNA-21,-221 and-222 were low in all groups and were not correlated with ischemic injury and/or chronic alcoholism. The serum expressions of miRNA-15b,-21,-221 and-222 were similar among the experimental groups, with no correlation with ischemia, with or without reperfusion, and/or alcoholism. The overexpression of miRNA-16 in the blood of I and IR groups demonstrated a correlation with the ischemic process, mainly after reperfusion for 48 hours, associated or not with alcoholism.
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