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The potential role of N6-methyladenosine modification of LncRNAs in contributing to the pathogenesis of chronic glomerulonephritis

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Abstract

Background

Increasing evidence indicates that N6-methyladenosine (m6A) modification of mRNAs has been shown to play a critical role in the occurrence and development of many diseases, while little is known about m6A modification in long non-coding RNAs (LncRNAs). Our study aims to investigate the potential functions of LncRNA m6A modifications in lipopolysaccharide (LPS)-induced mouse mesangial cells (MMCs), providing us with a new perspective on the molecular mechanisms of chronic glomerulonephritis (CGN) pathogenesis.

Methods

Differentially methylated LncRNAs were identified by Methylated RNA immunoprecipitation sequencing (MeRIP-seq). LncRNA–mRNA and LncRNA-associated LncRNA–miRNA–mRNA (CeRNA) networks were constructed by bioinformatics analysis. Furthermore, we utilized gene ontology (GO) and pathway enrichment analyses (KEGG) to explore target genes from co-expression networks. In addition, the total level of m6A RNA methylation and expression of methyltransferase and pro-inflammatory cytokines were detected by the colorimetric quantification method and western blot, respectively. Cell viability and cell cycle stage were detected by cell counting kit-8 (CCK-8) and flow cytometry.

Results

In total, 1141 differentially m6A-methylated LncRNAs, including 529 hypermethylated LncRNAs and 612 hypomethylated LncRNAs, were determined by MeRIP-seq. The results of GO and KEGG analysis revealed that the target mRNAs were mainly enriched in signal pathways, such as the NF-kappa B signaling pathway, MAPK signaling pathway, Toll-like receptor signaling pathway, and apoptosis signaling pathway. In addition, higher METTL3 expression was found in CGN kidney tissues using the GEO database. METTL3 knockdown in MMC cells drastically reduced the levels of m6A RNA methylation, pro-inflammatory cytokines IL6 and TNF-α, and inhibited cell proliferation and cycle progression.

Conclusions

Our findings provide a basis and novel insight for further investigations of m6A modifications in LncRNAs for the pathogenesis of CGN.

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Data availability

The data used and analyzed to support the findings of this study are available from the corresponding author upon request.

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Funding

This study was financially supported by the National Natural Science Foundation of China (No.81973546), the Key Scientific Research Projects of Natural Science in Colleges and Universities in Anhui Province (No.2022AH050747), and the Key Scientific Research Projects of Natural Science in Colleges and Universities in Anhui Province (No.2022AH050455).

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Author notes

  1. Tao Liu and Xing Xing Zhuang have contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmacy, The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei, 230012, Anhui, China

    Tao Liu, Xiu Juan Qin, Liang Bing Wei & Jia Rong Gao

  2. Department of Pharmacy, Chaohu Hospital of Anhui Medical University, Chaohu, 238000, Anhui, China

    Xing Xing Zhuang

  3. College of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230011, Anhui, China

    Tao Liu & Xing Xing Zhuang

  4. Anhui Province Key Laboratory of Chinese Medicinal Formula, Hefei, 230031, Anhui, China

    Jia Rong Gao

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JRG conceived and designed the study. TL and XXZ wrote the paper. JRG, XJQ, and BLW reviewed and edited the manuscript. All authors read and approved the manuscript.

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Correspondence to Jia Rong Gao.

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Liu, T., Zhuang, X.X., Qin, X.J. et al. The potential role of N6-methyladenosine modification of LncRNAs in contributing to the pathogenesis of chronic glomerulonephritis. Inflamm. Res. 72, 623–638 (2023). https://doi.org/10.1007/s00011-023-01695-2

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  • DOI: https://doi.org/10.1007/s00011-023-01695-2

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