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MicroRNA-339-5p inhibits lipopolysaccharide-induced rat mesangial cells by regulating the Syk/Ras/c-Fos pathway

Abstract

Chronic glomerulonephritis (CGN) is a disease occurred in glomeruli. The mechanism of CGN is regarded to be involved in a range of inflammatory responses. MicroRNA-339-5p (miR-339-5p) has been reported to be involved in inflammatory responses in many diseases. However, the role of miR-339-5p in CGN remains unclear. The purpose of this study was to investigate the role of miR-339-5p in lipopolysaccharide (LPS)-induced nephritis injury in vitro. The real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and western blot (WB) were used to detect the expression of miR-339-5p and Syk/Ras/c-Fos pathway. Double luciferase was performed to identify targeted binding of miR-339-5p to Syk. Cell counting kit-8 (CCK-8) and flow cytometry were used to observe cell viability and cell cycle. Enzyme-linked immunosorbent assay (ELISA) was performed to measure the concentrations of inflammatory cytokines IL-1β, IL-10, IL-6, and TNF-α. Lipopolysaccharide (LPS) could increase HBZY-1 (rat mesangial cells) cell viability, decrease the G2 phase, and promote cell proliferation and accelerate inflammatory cytokine. However, overexpression of miR-339-5p could inhibit LPS-induced HBZY-1 cell viability, decrease the expression of Syk/Ras/c-Fos signaling pathway, downregulate the expression level of inflammatory cytokines, increase the G2 phase, and inhibit cell proliferation. miR-339-5p could inhibit the proliferation and inflammation of the rat mesangial cells through regulating Syk/Ras/c-Fos signaling pathway.

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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 supported by the National Natural Science Foundation of China (NSFC)《Lnc NONRATG001910.2 competitively binds miR-339 to regulate the mechanism of Syk involvement in the pathogenesis of chronic nephritis and the intervention of Qiteng Xiaozhuo particles》(No.81973546).

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All authors contributed to the conception and design of this study. Material preparation, data collection, and analysis were performed by HJ, XLZ, XJQ, and LBW. The first draft of the manuscript was written by JRG and MMS and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript. The authors declare that all data were generated in-house and that no paper mill was used.

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Correspondence to Jia Rong Gao.

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Gao, J.R., Shi, M.M., Jiang, H. et al. MicroRNA-339-5p inhibits lipopolysaccharide-induced rat mesangial cells by regulating the Syk/Ras/c-Fos pathway. Naunyn-Schmiedeberg's Arch Pharmacol (2022). https://doi.org/10.1007/s00210-022-02261-z

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  • DOI: https://doi.org/10.1007/s00210-022-02261-z

Keywords

  • CGN
  • miR-339-5p
  • Syk/Ras/c-Fos
  • Proliferation