Dynamic Whole-Transcriptome landscape of Acute Bilirubin Encephalopathy in Newborns

To determine the dynamic expression of mRNA,miRNA, lncRNA, and circRNA within 24 hours after neonatal bilirubin-induced brain injury. The bilirubin encephalopathy model of neonatal SD rats was established by injecting bilirubin solution into cerebellar bulbar cisterna of 5-day-old SD rats. The acute bilirubin encephalopathy model(ABE) of newborn rats was established by injecting a bilirubin solution into the cisterna magna(CM) of 5-day-old Sprague-Dawley(SD) rats. The ABE models were divided into three groups based on the duration of bilirubin action: 6 hours, 12 hours, and 24 hours. The neurobehavioral manifestations of each group were observed. HE staining and Tunel assay were used to evaluate the pathological damage in the hippocampus of newborn rats. Newborn rats injected with an equal volume of ddH2O in the CM served as the control group. Isoflurane inhalation anesthesia was administered prior to all surgical procedures. Before the CM injection, an equal volume of cerebrospinal fluid was drawn to avoid intracranial hypertension. The rats were sacrificed at the corresponding time periods. Dissecting the brain and isolating two hippocampal tissues on a cold, sterile petri dish. snap-frozen using liquid nitrogen, and stored at -80 degree celsius until analysis.Two hippocampal tissues were dissected and isolated on a cold sterile petri dish, rapidly frozen using liquid nitrogen, and stored at -80 degree celsius until analysis. Total RNA was extracted from hippocampal lobes using TRIzol (Invitrogen, USA). Each group(Con, BE6h, BE12h, BE24h) consisted of five rats. The RNA samples underwent further processing for sequencing.