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Journal of Peking University (Health Sciences) ›› 2025, Vol. 57 ›› Issue (2): 237-244. doi: 10.19723/j.issn.1671-167X.2025.02.003

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Expression and regulatory mechanism of miR-34a in neonatal rat model of bron-chopulmonary dysplasia induced by hyperoxia

Mengyue HUO, Hua MEI*(), Yuheng ZHANG, Yanbo ZHANG, Chunli LIU   

  1. Department of Neonatology, Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010050, China
  • Received:2022-03-22 Online:2025-04-18 Published:2025-04-12
  • Contact: Hua MEI E-mail:meihuayani@sina.com
  • Supported by:
    the Natural Science Foundation of Inner Mongolia Autonomous Region Project(2020MS08034);the Major Scientific Research Project of Inner Mongolia Medical University Affiliated Hospital(NYFY ZD008)

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Abstract:

Objective: To investigate the expression and possible regulatory mechanism of miR-34a in the lung tissue of neonatal rat model of bronchopulmonary dysplasia (BPD) induced by hyperoxia. Methods: In the study, 80 newborn SD rats were randomly divided into hyperoxia group (FiO2=60%) and air group (FiO2=21%) within 2 hours after birth, 40 rats per group. Lung tissue samples of the SD rats in each group were extracted on the 1st, 7th, 14th and 21st days after birth, and the pathological changes of lung tissue were observed under light microscope after HE staining. The number of radial alveolar counts (RAC) and the mean alveolar diameter (MAD) and the thickness of alveolar septal thickness (AST) were measured to evaluate the development of alveoli. Real-time fluorescence quantitative PCR was used to detect the expression of miR-34a, angiopoietin-1 (Ang-1) and tyrosine kinase receptor-2 (Tie-2) in lung tissue of rats in hyperoxia group and air group at different time points. Enzyme-linked immunosorbent assay (ELISA) was used to detect the proteins expression of Ang-1 and Tie-2 in the lung tissues of the two groups at different time points. Results: The weight of rats in the hyperoxia group on the 7th, 14th and 21st days after birth was significantly lower than that in the air group (P all < 0.05). With the prolongation of oxygen exposure, the number of alveoli decreased, the volume increased, the structure simplified, the alveolar cavity enlarged obviously and the alveolar septum thickened in the hyperoxia group. On the 7th, 14th and 21st days after birth, the RAC in the hyperoxia group was significantly lower than that in the air group (P all < 0.05). Compared with the air group, MAD and AST increased significantly on the 7th, 14th and 21st days after birth in the hyperoxia group, and the difference was statistically significant (P all < 0.05). The expression level of miR-34a in lung tissue of hyperoxia group was significantly higher than that of air group on the 7th, 14th and 21st days after birth, and the difference was statistically significant (P all < 0.05). Compared with the air group at the same time point, the expression levels of Ang-1 and Tie-2 mRNA and protein in the hyperoxia group were lower than those in the air group on the 14th and 21st days after birth (P all < 0.05). Conclusion: The new BPD model of newborn SD rats can be successfully established by continuous exposure to 60% hyperoxia. The expression of miR-34a was up-regulated in the lung tissue of the new BPD model of neonatal rats. MiR-34a may play an important role in the occurrence and development of BPD by regulating Ang-1/Tie-2 signal pathway.

Key words: Bronchopulmonary dysplasia (BPD), miR-34a, Angiopoietin-1 (Ang-1), Tyrosine kinase receptor-2 (Tie-2), Hyperoxia

CLC Number: 

  • R722.1

Table 1

Primer sequences for quantitative real-time PCR"

Primer Sequence
miR-34a Forward 5′-CTCAACTGGTGTCGTGGA-3′
Reverse 5′-TCGGCAGGAATCAGCA-3′
Ang-1 Forward 5′-GGAACCGAGCCTACTCACAG-3′
Reverse 5′-ATCAGCGTCCTTTGTGCTGA-3′
Tie-2 Forward 5′-TTCACCAGGCTGATTGTCCG-3′
Reverse 5′-TCCTCCCCATAAACCCAGGA-3′

Figure 1

Weight changes of rats in the normoxia group and hyperoxia group at different time points *P < 0.05, compared with the normoxia group at the same time point; #P < 0.05, compared with the first day of this group; △P < 0.05, compared with the 7th day of this group; ☆P < 0.05, compared with the 14th day of this group."

Figure 2

Pathological changes in lung tissue of rats in the normoxia group and hyperoxia group at different time points (HE staining ×100) A to D, normoxia group on the 1st, 7th, 14th and 21st days; E to H, hyperoxia group on the 1st, 7th, 14th and 21st days."

Figure 3

Comparison of RAC(A), MAD(B) and AST(C) between normoxia group and hyperoxia group rats at different time points *P < 0.05, compared with the normoxia group at the same time point; # P < 0.05, compared with the first day of this group; △P < 0.05, compared with the 7th day of this group; ☆P < 0.05, compared with the 14th day of this group. RAC, radial alveolar counts; MAD, mean alveolar diameter; AST, alveolar septal thickness."

Figure 4

Amplification(A) and dissolution(B) curves of miR-34a"

Figure 5

Expression of miR-34a in lung tissue of rats in the normoxia group and hyperoxia group at different time points *P < 0.05, compared with the normoxia group at the same time point; #P < 0.05, compared with the first day of this group; △P < 0.05, compared with the 7th day of this group."

Figure 6

Expression of Ang-1 mRNA(A) and Tie-2 mRNA(B) in lung tissue of rats in the normoxia group and hyperoxia group at different time points *P < 0.05, compared with the normoxia group at the same time point; # P < 0.05, compared with the first day of this group; △P < 0.05, compared with the 7th day of this group; ☆P < 0.05, compared with the 14th day of this group."

Figure 7

Expression of Ang-1(A) and Tie-2(B) proteins in lung tissue of rats in the normoxia group and hyperoxia group at different time points *P < 0.05, compared with the normoxia group at the same time point; #P < 0.05, compared with the first day of this group; △P < 0.05, compared with the 7th day of this group; ☆P < 0.05, compared with the 14th day of this group."

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