ABSTRACT
The chronic consumption of alcohol causes a worsening of the events that follow the cerebral ischemia. These events are regulated through the expression of several genes and microRNAs. The aimof this work was To analyze and describe the expression profile of PARP and AIF and miRNA-9 proteins in rats submitted to focal cerebral ischemia, associated or not with chronic alcoholism model. Methods: Twenty adult Wistar rats, subdivided into: control; ischemic; alcoholic and ischemic / alcoholized for immunohistochemical analysis and miRNA-9 gene expression. Results: There was a reduction in the protein expression of PARP-1 and a positive marking for AIF in the ischemic / alcoholized group. The miRNA-9 did not obtain significant expression. The association of ischemia with chronic alcohol use promoted a tendency to low expression of miRNA-9, low expression of PARP-1 and high expression of AIF, indicating an interference in the protective effect of miRNA-9 be observed in the other groups.
El consumo crónico de alcohol provoca un empeoramiento de los eventos que siguen a la isquemia cerebral. Estos eventos están regulados a través de la expresión de varios genes y microRNA. El objetivo de este trabajo fue analizar y describir el perfil de expresión de las proteínas PARP y AIF y microRNA-9 en ratas sometidas a isquemia cerebral focal, asociadas o no, con el modelo de alcoholismo crónico. Veinte ratas Wistar adultas se dividieron en: grupo control, isquémico alcohólico, e isquémico / alcoholizado para análisis inmunohistoquímico y expresión de genes microRNA-9. Resultados: Hubo una reducción en la expresión de proteínas de PARP-1 y un marcado positivo para AIF en el grupo isquémico / alcoholizado. No se observó una expresión significativa en el microRNA-9. La asociación de la isquemia con el consumo crónico de alcohol promovió una tendencia a la baja expresión de microRNA-9, baja expresión de PARP1 y alta expresión de AIF, lo que indica una interferencia en el efecto protector de microRNA-9 en los otros grupos.
Subject(s)
Animals , Rats , Brain Ischemia/metabolism , Alcoholism/metabolism , Immunohistochemistry , Brain Ischemia/genetics , Rats, Wistar , MicroRNAs/metabolism , Disease Models, Animal , Alcoholism/genetics , Apoptosis Inducing Factor/metabolism , Poly (ADP-Ribose) Polymerase-1/metabolismABSTRACT
Aim: To explore the mechanism of osthole regulating the expression of miR-9 to delay the occurance of Alzheimer' s disease (AD). Methods: The miRNA which expressed differently with osthole treatment was selected by microarray; the target gene binding to miRNA-9 was verified by databases and Cytoscape; the SH-SY5Y cells with over expression of APP were established using Lipofectamine2000. The cell viability was determined by MTT assay. The miRNA-9 inhibitor was transfected into cells, and the expression of miRNA-9 and BACE-1 mRNA was determined by RT-PCR; the changes of BACE-1 protein were determined by Western blot. Results: The results of database showed that osthole-mediated miRNA-9 was combined with BACE-1 genes. Our lab had established SH-SY5Y cells with over expression of APP. The results of MTT assay showed that 50 p,mol · L-1 osthole had a protective effect on cells. Osthole could increase the expression of miRNA-9, and the expression of miR- 9 was lowest in inhibitor group determined by RT- PCR. Osthole decreased the expression of BACE-1 mRNA and protein compared with APP group, and the expression of BACE-1 was highest in inhibitor group. Conclusion: Osthole plays a protective role in AD partly through suppressing the expression of BACE-1 by up-regulating miRNA-9.