Silencing of Long Non-Coding RNA X Inactive Specific Transcript (Xist) Contributes to Suppression of Bronchopulmonary Dysplasia Induced by Hyperoxia in Newborn Mice via microRNA-101-3p and the transforming growth factor-beta 1 (TGF-β1)/Smad3 Axis

Abstract

Background Bronchopulmonary dysplasia (BPD) is a chronic lung disease mostly affecting premature infants. Long non-coding RNA (lncRNA) X inactive specific transcript (Xist) is actively involved in pulmonary disease development. The present study explored the potential mechanism of Xist in BPD development. Material Methods First, newborn BPD mouse models were successfully established. lncRNAs and genes with differential expression were identified using microarray analysis. Various injuries and radial alveolar counts of lung tissues of BPD mice were detected by hematoxylin-eosin staining. Functional assays were utilized to detect alterations of superoxide dismutase (SOD), malondialdehyde (MDA), vascular endothelial growth factor, collagen I, alpha-smooth muscle Actin, TGF-β1, and Smad3. Then, dual-luciferase reporter gene assay and RNA pull-down assay were performed to clarify the targeting relationship between Xist and miR-101-3p and between miR-101-3p and high-mobility group protein B3 (HMGB3). Results In BPD mice, radial alveolar counts value and SOD activity declined while MDA level increased. Results of microarray analysis found that Xist and HMGB3 were highly expressed in BPD mice. Next, silenced Xist alleviated lung damage in BPD mice. Xist competitively bound to miR-101-3p to activate HMGB3, and overexpressed miR-101-3p mitigated lung damage in BPD mice. Additionally, silenced Xist downregulated the TGF-β1/Smad3 axis. Conclusions Our study demonstrated that silencing of Xist suppressed BPD development by binding to miR-101-3p and downregulating HMGB3 and the TGF-β1/Smad3 axis. Our results may provide novel insights for BPD treatment.

Comprehensive Analysis of Long Non-coding RNA,miRNA，and mRNA-associated Competitive Endogenous RNA Networks in the Peripheral Blood of Preterm Infants with Bronchopulmonary Dysplasia

“…In addition, (36) found that SNHG6 can mediate lung cell damage by regulating the miR-335/KLF5/NF-κB pathway. The mechanism may be that Xist competitively binds miR-101-3p to activate HMGB3, and overexpression of miR-101-3p alleviates lung injury in BPD mice (37). lncRNA MALAT1 inhibits apoptosis and plays a protective role in BPD through the Keap1/Nrf2 signaling pathway (38).…”

Umbilical cord blood–derived exosomes from healthy term pregnancies protect against hyperoxia‐induced lung injury in mice

“…This is not surprising because a variety of miRNAs are known to affect the BPD development in rodent models. In particular, it has been shown that miR‐421 negatively regulate the BPD development in hyperoxia‐insulted mice 22 ; whereas overexpression of miR‐101‐3p inhibits lung injuries in BPD mice 23 . In addition, both miR‐451 and miR‐203a‐3p actively participate in the BPD development in mice 24 and rats, 25 partially via regulating vascular growth and function increase.…”

“…In particular, it has been shown that miR‐421 negatively regulate the BPD development in hyperoxia‐insulted mice 22 ; whereas overexpression of miR‐101‐3p inhibits lung injuries in BPD mice. 23 In addition, both miR‐451 and miR‐203a‐3p actively participate in the BPD development in mice 24 and rats, 25 partially via regulating vascular growth and function increase. We have previously revealed that miR‐185‐5p, which is lower in UCB‐EXO from BPD versus non‐BPD preterm pregnancies, promote endothelial angiogenesis in vitro.…”

LncRNA XIST promotes neovascularization in diabetic retinopathy by regulating miR-101-3p/VEGFA

“…It is well-acknowledged that lncRNAs can interact with microRNAs (miRNAs/miRs) to regulate downstream mRNA and gene expression ( 14 ). XIST targets miR-101-3p and inhibits its expression in silicosis, cancers, and bronchopulmonary dysplasia ( 15 - 18 ). Interestingly, miR-101-3p can repress vascular endothelial growth factor A (VEGFA) expression by targeting VEGFA mRNA, thus reducing cell migration and invasion ( 19 ).…”