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

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. vascular mRNA profiles of 3 SD rats after ischemia 1h and 3 SD rats after ischemia 4h and 3 SD rats for normal control were generated by deep sequencing using Hiseq Xten