Long noncoding RNA colorectal neoplasia differentially expressed alleviates sepsis-induced liver injury via regulating miR-126-5p

Abstract

In this study, we intended to determine the detailed function and mechanism of long noncoding RNA (lncRNA) colorectal neoplasia differentially expressed (CRNDE) in liver injury induced by sepsis. Cecal ligation and perforation (CLP) models were adopted to induce sepsis in vivo with rats, and hepatic epithelial cells L02 were treated with lipopolysaccharide (LPS) to mimic sepsis in vitro. Enzyme-linked immunosorbent assay was conducted to detect the levels of tumor necrosis factor (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), and interferon-γ (IFN-γ) in the serum of rats. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to measure the expressions of CRNDE and microRNA-126-5p (miR-126-5p). Flow cytometry analysis and Cell Counting Kit-8 (CCK-8) method were carried out followed by the up- or downregulation of CRNDE and miR-126-5p to monitor the proliferation and apoptosis of L02 cells, respectively. Western blot was then applied to determine the expressions of cysteinyl aspartate specific proteinase 3 (caspase 3), poly(ADP-ribose)polymerase (PARP), cytochrome c, and BCL2-like 2 (BCL2L2). The interactions between CRNDE with miR-126-5p and miR-126-5p with BCL2L2 were determined through bioinformatics, qRT-PCR, dual luciferase reporter assay, and RNA immunoprecipitation assay. CRNDE was significantly decreased in liver tissues and hepatic cells in sepsis models. Upregulation of CRNDE promoted the viability of L02 cells and inhibited their apoptosis, while downregulation of CRNDE had opposite effects. The expression of CRNDE in liver tissues of septic rats was correlated with the expression miR-126-5p. It was also demonstrated that the transfection of miR-126-5p mimics reversed the inhibitory effect induced by CRNDE on apoptosis of L02 cells. CRNDE could specifically bind to miR-126-5p and reduce its expression, in turn promote the expression of BCL2L2. Additionally, CRNDE overexpression in rats ameliorated liver injury induced by sepsis. Downregulated CRNDE aggravates hepatic injury via regulating miR-126-5p and BCL2L2 during sepsis.

Keywords: CRNDE; liver injury; miR-126-5p; sepsis.