Unknown,Transcriptomics,Genomics,Proteomics

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Temporal gene expression profiling of ischemic brain injury in rats after Semax and Pro-Gly-Pro treatment

ABSTRACT: A drug currently efficient for cerebral stroke therapy is Semax, a synthetic peptide bearing a fragment of ACTH (4â7) and the C-terminal tripeptide Pro-Gly-Pro (PGP) was included to ensure resistance to peptidases.The genome-wide expression changes induced by Semax and PGP in rat brain cortex tissues damaged by focal ischemia were studied using the genome-wide RatRef-12 Expression BeadChip (Illumina, USA), which contains 22,226 genes, according to NCBI. We compared the biochip data obtained at 3 h and 24 h after permanent middle cerebral artery occlusion (pMCAO) in each of the three groups (âischemia,â âischemia + Semax,â and âischemia + PGPâ). The transcriptome profiles were examined at 24 h vs. 3 h after pMCAO in rats that produced ischemic cortical injury and in rats with the same injury treated with Semax or PGP.

ORGANISM(S): Rattus norvegicus

SUBMITTER: Medvedeva Ekaterina

PROVIDER: E-MTAB-4658 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:TLR4/NF-ÎºB signaling plays a central mediator in response to danger signals released in the muscle ischemia-reperfusion injury (IRI). This study was designed to profile TLR4/NF-ÎºB-responsive microRNAs (miRNAs) in the skeletal muscles following IRI. Following 2 h of ischemia and subsequent reperfusion for indicated times (0 h, 4 h, 1 d, and 7 d) of the isolated thigh skeletal muscles based on femoral artery perfusion of C57BL/6, Tlr4â/â, and NF-ÎºBâ/âmice, the muscle specimens were analyzed with an miRNA array to detect the TLR4/NF-ÎºB-responsive miRNAs. Male C57BL/6 mice (10â12 weeks of age, 22â35 g) were purchased from BioLasco (Taiwan). Tlr4â/â (C57BL/10ScNJ) and NF-ÎºBâ/â (B6.Cg-Nfkb1tm1Bal/J) mice were purchased from Jackson Laboratory (Bar Harbor, ME, USA). All housing conditions were established and surgical procedures, analgesia, and assessments were performed in an AAALAC-accredited, SPF facility following national and institutional guidelines. Animal protocols were approved by the IACUC of Chang Gung Memorial Hospital. Mice were anesthetized with an anesthetic cocktail consisting of 0.1 mg/g ketamine and 0.01 mg/g xylazine, given as an intraperitoneal injection (0.01 ml/g body weight). After the induction of anesthesia, the mice were restrained in a supine position on a heating pad to maintain body temperature at 37Â°C. The quadriceps muscle perfused based on the femoral artery of the mouse was carefully isolated away from the femoral bone and the underlying adductor muscle group. In the ischemic group, ischemia was induced by placing a microvascular clamp carefully across the proximal site of vascular pedicle for 2 h and then the microvascular clamp was removed. Good vascular inflow and outflow through the pedicle was verified under direct magnified vision. In the sham-operated control group, the muscle was isolated without microvascular clamp being applied. The incision wound was closed with interrupted sutures (4-0 nylon) and the animals were allowed to awaken in the remaining periods of reperfusion. The harvested muscles were frozen in isopentane chilled in liquid nitrogen and stored at -80Â°C. For the miRNA array experiments, the isolated skeletal muscles of C57BL/6 mice after 2 h of ischemia and 0 h, 4 h, 1 d, and 7 d of reperfusion as well as of Tlr4â/âand NF-ÎºBâ/â mice after 2 h of ischemia and 1 d of reperfusion were used in 3 replicate experiments.

Project description:Experimental Design: 1. The goal of the experiment: Age-Dependent Gene Expression Profiles After Hepatic Ischemia/Reperfusion: Implications for the AP-1 and Ubiquitin-Proteosome Pathways 2. Brief description of the experiment: Hepatic ischemia/reperfusion (I/R) injury is a complication of liver surgery, transplantation and shock. Our previous studies have suggested there is an age-dependent response to I/R. In the present study, we examined global gene expression after liver I/R in young (4-5 weeks) and adult (12-14 months) mice using Affymetrix microarray. Gene expression was filtered based on a change in expression of 1.5-fold relative to respective controls and then analyzed by ANOVA. Seventy-two genes in young mice and 56 genes in adult mice had significantly increased expression. Of these, only 18 were up-regulated in both age groups. In young mice, 289 genes were down-regulated whereas in adult mice, 874 genes were decreased. Of these genes, 175 genes were decreased in both groups. Pathway and network analyses of up- and down-regulated gene lists revealed a number of notable differences between young and adult mice. Of these, we found that genes related to the activating protein-1 (AP-1) pathway were upregulated preferentially in young mice. This corresponded with an increase in AP-1 activation in young versus adult mice. Finally, we found that genes related to the ubiquitin-proteasome pathway were selectively down-regulated in adult mice. This was accompanied by reduced degradation of the inhibitory protein, I?B?, in adult mice. The data demonstrate specific, pathway- and network-related differences in gene expression profiles between young and adult mice in the response to I/R. More specifically, we have identified two pathways that may contribute to the superior response to I/R in young mice which represent potential therapeutic targets. Keywords: treated vs non treated 2 age groups of 6 young and 6 adult mice that underwent either 90 minutes of partial ischemia followed by 1 hour of reperfusion (n=3) or sham operation (n=3) We used microarray to uncover the genomic response after ischemia reperfusion in 2 different age groups

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Temporal gene expression profiling of ischemic brain injury in rats after Semax and Pro-Gly-Pro treatment

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