miR-128-3p inhibits apoptosis and inflammation in LPS-induced sepsis by targeting TGFBR2

Peng Yang; Jianhua Han; Shigeng Li; Shaoning Luo; Xusheng Tu; Zhiqiang Ye

doi:10.1515/med-2021-0222

miR-128-3p inhibits apoptosis and inflammation in LPS-induced sepsis by targeting TGFBR2

Open Med (Wars). 2021 Feb 4;16(1):274-283. doi: 10.1515/med-2021-0222. eCollection 2021.

Authors

Peng Yang¹, Jianhua Han², Shigeng Li², Shaoning Luo², Xusheng Tu², Zhiqiang Ye²

Affiliations

¹ Department of Anesthesiology, The First Affiliated Hospital, Sun Yat-sen University, 510080, Guangzhou, China.
² Department of Emergency, The Third Affiliated Hospital, Sun Yat-sen University, No. 600 Tianhe Road, Tianhe District, 510630, Guangzhou, China.

Abstract

Background: Sepsis is a systemic inflammatory response that can lead to the dysfunction of many organs. The aberrant expression of miRNAs is associated with the pathogenesis of sepsis. However, the biological functions of miR-128-3p in sepsis remain largely unknown, and its mechanism should be further investigated. This study aimed to determine the regulatory network of miR-128-3p and TGFBR2 in lipopolysaccharide (LPS)-induced sepsis.

Methods: The expression levels of miR-128-3p and transforming growth factor beta receptors II (TGFBR2) were detected by quantitative polymerase chain reaction (qPCR). The protein levels of TGFBR2, Bcl-2, Bax, cleaved caspase 3, Smad2, and Smad3 were measured by western blot. Cell apoptosis was analyzed by flow cytometry. Cytokine production was detected by enzyme-linked immunosorbent assay (ELISA). The binding sites of miR-128-3p and TGFBR2 were predicted by Targetscan online software and confirmed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay.

Results: The level of miR-128-3p was decreased, and TGFBR2 expression was increased in serum samples of sepsis patients and LPS-induced HK2 cells. Overexpression of miR-128-3p or knockdown of TGFBR2 ameliorated LPS-induced inflammation and apoptosis. Moreover, TGFBR2 was a direct target of miR-128-3p, and its overexpression reversed the inhibitory effects of miR-128-3p overexpression on inflammation and apoptosis in LPS-induced HK2 cells. Besides, overexpression of miR-128-3p downregulated TGFBR2 to suppress the activation of the Smad signaling pathway.

Conclusion: miR-128-3p could inhibit apoptosis and inflammation by targeting TGFBR2 in LPS-induced HK2 cells, which might provide therapeutic strategy for the treatment of sepsis.

Keywords: TGFBR2; apoptosis; inflammation; miR-128-3p; sepsis.