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Transcriptome Analysis of rat vascular molecular pathways linked to mitochondria in different phase of ischeia

ABSTRACT: Purpose: to determine the molecular pathways linked to mitochondria in different phase of ischemia. Methods: Rats (220–240 g) were anaesthetized with sodium pentobarbital (initial dosage, 30 mg/kg). Anaesthetized rats were placed on a warmed plate to maintain the body temperature at 37°C. Aseptic techniques were adopted for all surgical procedures. Right femoral arteries were catheterized with polyethylene catheters for bleeding 50% of total blood volume (≈7% of weight). The ischemia time started to calculate after the model was established. At the end of ischemia, rats were killed with a lethal dose of sodium pentobarbital (100 mg/kg, iv). A laparotomy was then carried out to obtain superior mesenteric artery tissues (SMAs). Results: Base on the Mitochondrial-Genome Microarrays and GSEA database, 1200 DEGs were picked out among the 46095 test probes on the chips. The top 10 mitochondrial-DEGs include Gdap1, Inf2, Pdzd8, Bnip3, Bnip3l, Tomm20, Bik, Lrrk2, Bcl2l1, Ubb. We also find that mitochondrial morphology variation, including mitochondrial fission and fusion, and the interaction between endoplasmic reticulum (ER) and mitochondria were involved in early stage of ischemia, while mitochondrial dysfunctions occurred in late stage of ischemia, including the release of CytC, mitochondrial membrane potential variation and changes in mitochondrial membrane permeability. Conclusions: variation in mitochondrial morphology and dynamics occurs prior to mitochondrial dysfunctions after ischemic injury.

ORGANISM(S): Rattus norvegicus

PROVIDER: GSE123542 | GEO | 2020/04/29

REPOSITORIES: GEO

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Project description:Although disruption of mitochondrial function has been associated with energetic deregulation in cancer, the chronological changes in mitochondria during cancer development remain unclear. With the aim to assess the role of mitochondria throughout cancer development, we analyzed samples chronologically obtained from induced hepatocellular carcinoma (HCC) in rats. In our analyses, we integrated mitochondrial proteomic data, mitochondrial metabolomic data and nuclear genome transcriptomic data. We used pathway over-representation and weighted gene co-expression network analysis (WGCNA) to integrate expression profiles of genes, miRNAs, proteins and metabolite levels throughout HCC development. Our results show that mitochondria are dynamic organelles presenting specific modifications in different stages of HCC development. For carcinogenesis, a modified resistant hepatocyte model was used. Rats were initiated with diethylnitrosamine (DEN) (200 mg/kg of body weight) at day 0. Then, 2-acetylaminofluorene (AAF) was administered (20 mg/kg per dose) at days 7, 8 and 9, followed by 3/5 partial hepatectomy (PH) at day 10. Three control groups of non-treated animals were sacrificed by exsanguination on the first day and at 9 and 18 months after the beginning of the experiment. Treated animals were sacrificed at 1, 7, 11 and 16 days and at 1, 9 and 18 months. Their livers were excised, washed in physiological saline solution, frozen with liquid nitrogen in 2-methyl butane and stored at -80°C. Once selected and separately collected nodular, tumoral and its adjacent tissues, total RNA was isolated using TriPureIsolation Reagent (Roche) according to the manufacturer’s protocol. The microarray analysis was performed using GeneChip Rat Exon 1.0 ST Arrays. Four replicas for each condition and controls were analyzed. This created 11 pairwise contrasts for the differential expression analysis. We grouped all evaluated conditions in three categories: i) early stages, including samples obtained since day one until one month after treatment, these were compared against rats without treatment sacrificed the first day; ii) tumor, nodular and its adjacent tissues obtained from nine months rats were compared against rats without treatment of the same age; and iii) tumor and its adjacent tissues obtained from eighteen months rats were compared against rats without treatment of the same age.

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Transcriptome Analysis of rat vascular molecular pathways linked to mitochondria in different phase of ischeia

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