Uterines RNA-seq Analysis in Rats with Embryo Implantation Dysfunction
This study aimed to compare the transcriptomic changes in uterine tissue of rats with embryo implantation dysfunction (EID) before and after treatment with Jiawei Shoutai Wan (JWSTW). JWSTW is composed of seven medicinal herbs: Tu Si Zi, E Jiao, Sang Ji Sheng, Xu Duan, Dan Shen, Dang Gui, and Xiang Fu. Seven-week-old Sprague-Dawley (SD) rats were used in the experiment and housed in a specific pathogen-free (SPF) animal facility. The rat EID model was established by subcutaneous injection of a mifepristone oil solution into the neck and back region. After one week of acclimatization, female rats with regular estrous cycles were selected. At 5:00 PM daily, female and male rats were caged at a ratio of 2:1. Vaginal smears were collected from the female rats at 7:00 AM the following morning, and the presence of spermatozoa in the smear was designated as gestation day 1 (GD1). The mated female rats were randomly divided into a model group (administered an equal volume of normal saline via gavage) and a JWSTW group (administered 8.4 g/kg JWSTW via gavage). All female rats were administered mifepristone on the morning of GD4 to establish the model. From GD1 to GD6, gavage administration was performed daily at 9:00 AM. One hour after gavage on GD6, the rats were anesthetized by intraperitoneal injection of sodium pentobarbital, and uterine tissue was collected under aseptic conditions. Total RNA was extracted from the rat uterine tissue using the Trizol method, and its quality was assessed with the Agilent 2200 TapeStation. Following mRNA purification, reverse transcription was performed to synthesize cDNA, and libraries were subsequently constructed. The constructed libraries passed quality control and were sequenced on the Illumina HiSeq platform. Gene Ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using the clusterProfiler R package. Finally, Gene Set Enrichment Analysis (GSEA) was carried out.