Chunlei Liu,1,2 Changyuan Yue,1,2 Xiaoxiang Xing,1,2 Lidong Huang,1,2 Yanxin Wei,1,2 Peng Zhao,1,2 Jiansheng Li,1,3 Qingzhou Guan1 

1Henan Key Laboratory of Chinese Medicine for Respiratory Disease, Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-Constructed by Henan Province and Education Ministry of People’s Republic of China, Henan University of Chinese Medicine, Zhengzhou, 450046, People’s Republic of China; 2Academy of Chinese Medical Sciences, Henan University of Chinese Medicine, Zhengzhou, 450046, People’s Republic of China; 3Department of Respiratory Diseases, The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou, 450000, People’s Republic of China

Correspondence: Qingzhou Guan, Email a5410980@163.com

Objective: To investigate the mechanism by which the effective-component combination of Bufei Yishen formula III (ECC-BYF III) ameliorates airway epithelial barrier injury in chronic obstructive pulmonary disease (COPD) through miRNA-mRNA regulatory networks.
Methods: Differentially expressed mRNAs (DEmRNAs) and miRNAs (DEmiRNAs) were identified using the edgeR algorithm. The target genes of DEmiRNAs were predicted using four online databases. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis based on the hypergeometric distribution model was performed for DEmRNAs and the predicted target genes, respectively. DEmiRNAs and their corresponding target genes that were regulated by ECC-BYF III were subsequently identified. The reliability of these miRNAs and target genes was validated using independent datasets, qRT-PCR in human bronchial airway epithelial cells (BEAS-2B), COPD rat models, and molecular docking.
Results: Compared with the control group, 2997 DEmRNAs and 4 DEmiRNAs were identified in the model group (edgeR, P< 0.05). A total of 2430 target genes of the DEmiRNAs were predicted, and the miRNA-mRNA regulatory network was constructed. Pathway enrichment analysis revealed that DEmRNAs were enriched in 96 pathways and target genes in 112 pathways, with 53 overlapping pathways (P< 0.05). ECC-BYF III treatment reversed the expression of 13 miRNA-mRNA pairs. Further screening and validation in COPD rat models and BEAS-2B cells identified two miRNAs and five regulated hub genes (ESM1, RNF44, BCL2L1, ADAM19, and SMYD5). The reliability of these hub genes was further confirmed by independent datasets (GSE173896 and GSE11906) and molecular docking.
Conclusion: ECC-BYF III may alleviate airway epithelial barrier injury in COPD by regulating the hsa-miR-3685-ESM1 and hsa-miR-3936-RNF44/BCL2L1/ADAM19/SMYD5 networks.

Keywords: chronic obstructive pulmonary disease, airway epithelial barrier injury, ECC-BYF III, miRNA-mRNA regulatory network