Illumina HiSeq 2500 sequencing; RNA-seq of  non-ischemic heart tissues treated with a traditional Chinese medicine formula (BYD) against myocardial ischemia in Sprague Dawley rats
RNA-seq of  non-ischemic heart tissues treated with a traditional Chinese medicine formula (BYD) against myocardial ischemia in Sprague Dawley rats
BYD is a classic traditional Chinese medicine consists of the water extraction of astragalus root, ginseng, processed licorice, and cinnamon at a ratio of 6:2:2:1. Under the deeply anaesthetized condition after echocardiographic examinations, all rats were sacrificed for collecting the  heart tissues samples. The myocardial tissues of the ischemic LV and the peri-ischemic rim (an approximately 1 mm rim of normal-appearing tissues) were removed, and the remaining tissues that consisted of non-ischemic LV were obtained. First, the widely used model produced by the occlusion of the left anterior descending coronary artery (LADCA) was employed to induce Myocardial Ischemia in rats. Prior to the operation, the rats were fasted overnight with free access to water. Briefly, an anterior thoracotomy was performed under sterile conditions to open the pericardium and the heart was then rapidly exteriorized. The LADCA was ligated approximately 2‒3 mm distal from its origin with the use of a 5-0 polypropylene suture. After surgery, the rats in the sham and model group received the same volume of saline vehicle and the rats in the TCM group received a dose of BYD at 1460mg/kg for one week. The RNA integrity was analyzed on a 1% agarose gel. Beads that contained oligo (dT) were used to isolate the poly mRNA from the total RNA and then the rRNA was subsequently removed. The first strand of cDNA was synthesized by using Random Hexamers and the second strand of cDNA was synthesized by adding buffer, dNTPs, RNase H and DNA Polymerase I, then purified by QIAQuick PCR kit and eluted with EB buffer. The purified double-stranded cDNA is then subjected to end-repair, base A addition and sequencing, followed by agarose gel electrophoresis to recover the target fragment and perform PCR amplification to complete the entire library preparation.
Experimental Factor: compound
BYD
Experimental Factor: dose
1460
Experimental Factor: injury
occlusion of the left anterior descending coronary artery