strain: Sprague-Dawley gender: male treated with: contrast medium tissue: plasma
Treatment protocol
male Sprague-Dawley rats (250-300g) were deprived of water for 3 days and then given furosemide by intramuscularly injection at a dose of 10 mg/kg. For CI-AKI group, a nonionic low-osmolar contrast medium, Omnipaque (350 mg I/ML, GE Healthcare, Shanghai, China) at a dose of 10 ml/kg, was subsequently administered via tail vein over the course of 5 min. For control group, the same amount of normal saline was given. Peripheral blood and kidney samples were harvest at 8h after contrast medium/normal saline administration.
Growth protocol
standard rat chow, specific pathogen free
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted and purified using mirVana™ miRNA Isolation Kit (Cat#AM1560, Ambion, Austin, TX, US) following the manufacturer’s instructions and checked for a RIN number to inspect RNA integration by an Agilent Bioanalyzer 2100(Agilent technologies, Santa Clara, CA, US)
Label
biotin
Label protocol
miRNA molecular in total RNA was labeled by miRNA Complete Labeling and Hyb Kit (Cat # 5190-0456, Agilent technologies, Santa Clara, CA, US) followed the manufacturer’s instructions, labeling section.
Hybridization protocol
Each slide was hybridized with 100ng Cy3-labeled RNA using miRNA Complete Labeling and Hyb Kit (Cat # 5190-0456, Agilent technologies, Santa Clara, CA, US) in hybridization Oven (Cat # G2545A, Agilent technologies, Santa Clara, CA, US) at 55°C，20 rpm for 20 hours according to the manufacturer’s instructions, hybridization section. After hybridization, slides were washed in staining dishes (Cat # 121, Thermo Shandon, Waltham, MA, US) with Gene Expression Wash Buffer Kit (Cat #5188-5327, Agilent technologies, Santa Clara, CA, US).
Scan protocol
Slides were scanned by Agilent Microarray Scanner (Cat # G2565CA, Agilent technologies, Santa Clara, CA, US) and Feature Extraction software 9.5.3 (Agilent technologies, Santa Clara, CA, US) with default settings.
Description
Eu miRNA profile in rat plasma
Data processing
Raw data were normalized by Quantile algorithm, Gene Spring Software 10.0 (Agilent technologies, Santa Clara, CA, US).
