Abstract
Objective To investigate the effects of lncRNA CRNDE on sepsis-associated acute kidney injury in the human kidney 2 cell line and explore the potential mechanisms. Methods HK-2 cells were treated with lipopolysaccharides to induce injuries. The expression of CRNDE and miR-146a in HK-2 cells were altered by a transient transfection assay. Cell apoptosis was detected by a flow cytometry assay, and the levels of inflammatory cytokines including TNF-α, IL-6, IL-8, and IL-1β were assessed by ELISA. Furthermore, western blot analysis was performed to detect the expression levels of TLR4/NF-κB pathway-related proteins. And a luciferase reporter gene assay was used to verify if miR-146a was the target of CRNDE. Results LPS treatment increased CRNDE expression in HK-2 cells. CRNDE overexpression enhanced cell injuries in HK-2 cells significantly increasing inflammatory cytokine levels, including TNF-α, IL-6, IL-8, and IL-1β, and cell apoptosis. In addition, CRNDE overexpression further activated the TLR4/NF-κB pathways in HK-2 cells. Inversely, opposite results were observed in the miR-146a mimic treatment group, and the miR-146a inhibitor could reverse the protein expression changes of TLR4/NF-κB in the si-CRNDE and LPS treatment group. Conclusion This study demonstrated that CRNDE overexpression could activate the TLR4/NF-κB signaling pathway by regulating miR-146a, which accelerated LPS-induced inflammation and apoptosis in HK-2 cells.
Highlights
Sepsis is a systemic inflammatory response syndrome caused by bacterial (e.g., Staphylococcus aureus), viral, or fungal infections, and it is the main cause of death in intensive care units (ICUs) [1]
We used LPS to stimulate HK-2 cells to construct the acute kidney injury (AKI) cell model. qRT-PCR results showed that CRNDE expression was significantly increased after LPS stimulated HK-2 cells (Figure 1)
These results suggest that LPS can upregulate CRNDE expression in HK-2 cells
Summary
Sepsis is a systemic inflammatory response syndrome caused by bacterial (e.g., Staphylococcus aureus), viral, or fungal infections, and it is the main cause of death in intensive care units (ICUs) [1]. LPS has been widely used in the establishment of an acute renal injury model in vitro [4, 5]. In mice and other animal models, LPS can induce acute inflammation by activating nuclear factor-kappa B (NF-kappa B) while inducing AKI [6]. NF-kappa B is an important transcription factor downstream of the endotoxin signal transduction pathway. When AKI occurs, an endotoxin induces cytokine production, accompanied by the activation of the NF-kappa B signaling pathway, which leads to a series of cascade reactions of cytokines (such as tumor necrosis factor-alpha, interleukin-1, and interleukin-6), i.e., inflammation, resulting in cell death, necrosis, and other injuries. Inhibiting the activation of the NF-kappa B signaling pathway may prevent the damage of AKI in sepsis
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