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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 / Análisis morfométrico y modelo de expresión de proteína y genes de apoptosis en isquemia cerebral focal en ratas y la acción neuroprotectora de hipotermia y ketoprofeno
Tirapelli, Daniela P. C; Cirino, Mucio Luiz de Assis; Carvalho, Camila A. Melo de; Novais, Paulo Cezar; Figueredo, Rafael Zimak; Porsani, Lucas Barbosa; Gula, Isabella Stracieri; Rodrigues, Andressa Romualdo; Silva, Jairo Pinheiro da; Pereira, Adriana Lis; Lizarte Neto, Fermino Sanches; Schimming, Bruno Cesar; Tazima, Maria de Fátima Galli Sorita; Fazan, Valéria de Paula Sassoli; Carlotti-Jr, Carlos Gilberto; Tirapelli, Luis Fernando; Colli, Benedicto Oscar.
  • Tirapelli, Daniela P. C; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Cirino, Mucio Luiz de Assis; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Carvalho, Camila A. Melo de; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Novais, Paulo Cezar; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Figueredo, Rafael Zimak; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Porsani, Lucas Barbosa; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Gula, Isabella Stracieri; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Rodrigues, Andressa Romualdo; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Silva, Jairo Pinheiro da; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Pereira, Adriana Lis; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Lizarte Neto, Fermino Sanches; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Schimming, Bruno Cesar; UNESP. Department of Anatomy. Botucatu. BR
  • Tazima, Maria de Fátima Galli Sorita; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Fazan, Valéria de Paula Sassoli; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Carlotti-Jr, Carlos Gilberto; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Tirapelli, Luis Fernando; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
  • Colli, Benedicto Oscar; USP. FMRP. Department of Surgery and Anatomy. Ribeirao Preto. BR
Int. j. morphol ; 38(3): 523-529, June 2020. graf
Article in English | LILACS | ID: biblio-1098282
Responsible library: CL1.1
ABSTRACT
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.
RESUMEN
Este estudio tuvo como objetivo investigar la morfometría y el patrón de expresión de proteínas y genes relacionados con la vía apoptótica extrínseca en la isquemia cerebral focal experimental y el agujero de neuroprotección con hipotermia y ketoprofeno. Se dividieron aleatoriamente 120 ratas en 3 grupos (20 animales cada uno) control - sin cirugía (20 animales); simulación - simulación de cirugía (20 animales); isquemia isquemia focal durante 1 hora, sin reperfusión (80 animales) y dividida en cuatro subgrupos con 20 animales cada uno isquemia + hipotermia intraisquémica; isquemia + ketoprofeno intravenoso previo, e isquemia + hipotermia y ketoprofeno. El volumen del infarto se midió utilizando un análisis morfométrico de áreas de infarto definidas por cloruro de trifenil tetrazolio y los patrones de expresión de los genes de apoptosis (Fas, c-Flip, caspase-8 y caspase-3) y la proteína de apoptosis caspase-3 fueron evaluados por PCR cuantitativa en tiempo real e inmunohistoquímica, respectivamente. Se observó hipoexpresión de genes de la vía extrínseca de la apoptosis receptor Fas, c-Flip y caspasa-8 en las áreas isquémicas. También se observaron aumentos en el gen y la proteína caspasa-3 en las áreas isquémicas y estos aumentos se redujeron por hipotermia y ketoprofeno, también observado por estudio morfométrico. El aumento de caspasas-3 sugiere que este gen tiene un papel importante en la apoptosis, y probable causa de muerte celular, involucrando el efecto neuroprotector de la hipotermia y el ketoprofeno.
Subject(s)


Full text: Available Index: LILACS (Americas) Main subject: Brain Ischemia Limits: Animals Language: English Journal: Int. j. morphol Journal subject: Anatomy Year: 2020 Type: Article Affiliation country: Brazil Institution/Affiliation country: UNESP/BR / USP/BR

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Full text: Available Index: LILACS (Americas) Main subject: Brain Ischemia Limits: Animals Language: English Journal: Int. j. morphol Journal subject: Anatomy Year: 2020 Type: Article Affiliation country: Brazil Institution/Affiliation country: UNESP/BR / USP/BR