MicroRNA miR-212-5p Regulates the MEK/ERK Signaling Pathway by Targeting A-Raf proto-oncogene serine/threonine-protein kinase (ARAF) to Regulate Cowshed PM2.5-Induced NR8383 Apoptosis

Ke Sun; Yize Sun; Yunna Jia; Xinran Duan; Zhenhua Ma; Xiqing Zhang; Lixia Wang; Yanbin Zhu; Yunhang Gao; Wangdui Basang

doi:10.3390/toxics11120981

MicroRNA miR-212-5p Regulates the MEK/ERK Signaling Pathway by Targeting A-Raf proto-oncogene serine/threonine-protein kinase (ARAF) to Regulate Cowshed PM_2.5-Induced NR8383 Apoptosis

Toxics. 2023 Dec 3;11(12):981. doi: 10.3390/toxics11120981.

Authors

Ke Sun^{1

2}, Yize Sun¹, Yunna Jia¹, Xinran Duan¹, Zhenhua Ma¹, Xiqing Zhang¹, Lixia Wang³, Yanbin Zhu², Yunhang Gao¹, Wangdui Basang²

Affiliations

¹ College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China.
² Institute of Animal Husbandry and Veterinary Medicine, Tibet Academy of Agricultural and Animal Husbandry Science, Lhasa 850009, China.
³ Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Changchun 130102, China.

Abstract

Objective: To investigate the role of miR-212-5p-targeted ARAF during the apoptosis of rat alveolar macrophages induced by cowshed PM_2.5. Methods: miRNA and related target genes and pathways were predicted using the KEGG, TargetScan, and other prediction websites. NR8383 macrophages were treated with cowshed PM_2.5 to establish an in vitro lung injury model in rats; meanwhile, for the assessment of cell viability, apoptosis, intracellular calcium ions, and mitochondrial membrane potential in NR8383 cells, RT-qPCR was used to detect the expression of miR-212-5p and the target gene ARAF. Results: The bioinformatic analyses showed that miR-212-5p and ARAF were involved in PM_2.5-associated cellular damage. Exposure to different concentrations (0 μg/mL, 60 μg/mL, 180 μg/mL, 300 μg/mL) with different durations (0 h, 12 h, 24 h, 48 h) of cowshed PM_2.5 resulted in apoptosis, increased intracellular calcium ions, and decreased mitochondrial membrane potential. The miR-212-5p mimic group showed an up-regulation of Bax and cleaved Caspase 3 expression but decreased Bcl2 expression compared to the NC group, and overexpression of ARAF up-regulated the expression of p-MEK1/2 and p-ERK1/2 and simultaneously reversed the above phenomena. Conclusions: miR-212-5p targets ARAF to affect the cowshed PM_2.5-induced apoptosis through the MEK/ERK signaling pathway, providing a potential target for relevant farming industry and pathology studies.

Keywords: MEK/ERK signaling pathway; apoptosis; cowshed PM2.5; miR-212-5p; mitochondrial damage.

Abstract

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