Preliminary Study On The Expression Profile And Function Of Long-Non-Coding RNA In Hyperoxia-Induced Rat BPD Model

Purpose:Bronchopulmonary dysplasia is a severe chronic lung disease in preterm.Heredity,infection and hyperoxia are involved in the pathogenesis,and there is no effective treatment;IncRNAs are key regulators of various biological processes.However,the mechanism of IncRNA in the development of BPD remains unclear.The purpose of this study was to explore the differential expression of IncRNA in hyperoxia-induced bronchopulmonary dysplasia model and to predict the function of lncRNA.In addiction,establish lncRNA and target gene network map for preliminary bioinformatics study,so to provide a theoretical basis for the study of BPD.Method:1.Newborn rats were randomly assigned to hyperoxia(85%O2)or the control group:the normoxia group(21%O2).Lung tissues were collected on days 1,3,7 and 14days.The BPD animal model was validated using HE staining,Masson staining,real-time RT-PCR and Western-blot.2.LncRNA library was constructed and tested,and deep sequencing was used to reveal the differential expression of long non-coding RNAs in hyperoxia bronchopulmonary dysplasia rat models.3.Nine target IncRNAs that may be related to BPD were confirmed by RT-PCR on the samples.4.KEGG,GO function and new transcript were predicted,variable shear events were analyzed,and a network diagram of lncRNA ceRNA related to TGF-beta and P53 pathway was constructed.Results:1.The lung tissue inflammatory manifestations were alleviated and taken over by pulmonary interstitia hyperplasia and fibrocyte proliferation after 14 days of hyperoxic exposure.Lung injury scores were compared on days 3,7,and 14 respectively,with significant differences.2.The expression of several BPD biomarkers was detected by RT-PCR and Western-blot,which was consistent with the literature reported.3.A total of 1175 different lincRNAs were detected in the hyperoxia group and the normoxia group,of which 544 were up-regulated and 631 were down-regulated.Differences of lncRNA distribution between the two groups were analyzed by scatter plot,heat map analysis,volcano chart analysis and lncRNA distribution on chromosomes.4.Nine lincRNA were validated in the sample.5.673 moleculars related to GO functions were enriched,including cell location and biological process.Pathway enrichment analysis showed that lncRNA was involved in 257 KEGG pathways.A network diagram of lncRNA ceRNA related of TGF-beta and p53 was constructed.Variable shear events were analyzed.9816 and 13098 new transcripts were predicted in hyperoxia group and in normoxia group respectively.Conclusion:1.We established a hyperoxia rats BPD model successfully.The neonatal SD rats inhaled high concentration of oxygen for 14 days can construct animal model of bronchopulmonary dysplasia.2.Long time hyperoxia changes the expression of IncRNA in BPD rat model,indicate lncRNA plays a role in the occurrence and development of BPD.3.RT-PCR analysis confirmed the reliability and accuracy of RNA-seq data.4.Many target genes were involved in the developmental process,including cell component biogenesis,biological regulation,transcription regulator,and translation regulator.The BPD might be caused by the activation of the pathways of the EMC-receptor interaction,cytokine-cytokine receptor interaction,cell cycle,and cell adhesion molecules.5.Variable shear analysis shows that IncRNA is involved in the regulation of transcription.Predicting new transcripts is an important complement of the BPD pathogenesis.The network diagram constructed by IncRNA related to TGF-beta and P53 pathway,suggests that its own and the pathway involved can become a new target for the treatment of bronchopulmonary dysplasia.