Long non-coding RNA PVT1 promote LPS-induced septic acute kidney injury by regulating TNFα and JNK/NF-κB pathways in HK-2 cells.

This study aimed to investigate the effect and underlying mechanism of long non-coding RNA plasmacytoma variant translocation 1 (PVT1) in lipopolysaccharide (LPS)-induced inflammation injury in HK-2 cells. We established LPS-induced septic acute kidney injury (AKI) model in HK-2 cells. LPS-induced HK-2 cells were transfected with pc-PVT1, pc-NC, si-PVT1 or si-NC. Cell viability and apoptosis rate were detected by MTT assay and Annexin V-FITC/PI Apoptosis Detection kit, respectively. The relationships of PVT1 and inflammatory factors were evaluated by RNA Immunoprecipitation (RIP) assay. The levels of inflammatory factors, apoptosis-related proteins and the expressions of proteins related to c-Jun N-terminal kinase (JNK) and nuclear factor-κB (NF-κB) signaling pathway were detected by ELISA or Western blotting. Compared with cells with pc-NC, cell viability was remarkably decreased and cell apoptosis rate was increased in LPS-induced cells with pc-PVT1 (p<0.05). The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β were significantly increased in LPS-induced cells with pc-PVT1 compared with cells with pc-NC (p<0.05). All these changes were reversed in LPS-induced cells with si-PVT1 and si-NC (p<0.05). RTP assay revealed that PVT1 could bind to TNF-α. Furthermore, down-regulated PVT1 remarkably reduced the expressions of p-JNK and p-c-Jun, p-IκBα and p-p65 (p<0.05); while increased expressions of these proteins and inflammatory factors induced by up-regulated PVT1 were reversed by JNK or NF-κB inhibitors. PVT1 may promote inflammatory response by binding to TNF-α and inhibiting JNK/NF-κB signaling pathway in LPS-induced septic AKI cells.

Huang W ，Lan X ，Li X ，Wang D ，Sun Y ，Wang Q ，Gao H ，Yu K ... -  《-》

[Scutellarin alleviates lipopolysaccharide-induced renal injury via mediating cysteine-rich protein 61-connective tissue growth factor-nephroblastoma overexpressed gene 1 expression to inhibit nuclear factor-κB signaling pathway].

Liu X ，Qin Z ，Guan C ，Xu L ，Dai J ，Yang C ，Zhou B ，Luan H ，Zhao L ，Zhang W ，Luo C ，Xu Y ... -  《-》

Klotho protects against LPS-induced inflammation injury by inhibiting Wnt and NF-κB pathways in HK-2 cells.

Zhou Y ，Kuang Y ，Zhou J 《PHARMAZIE》

Circ_0114427 promotes LPS-induced septic acute kidney injury by modulating miR-495-3p/TRAF6 through the NF-κB pathway.

Xu L ，Cao H ，Xu P ，Nie M ，Zhao C ... -  《-》

LncRNA PVT1 accelerates LPS-induced septic acute kidney injury through targeting miR-17-5p and regulating NF-κB pathway.

Long noncoding RNA PVT1 is associated with diverse human diseases, including acute kidney injury (AKI). However, our understandings of PVT1 on septic AKI are limited. The septic AKI model was constructed through lipopolysaccharide (LPS) treatment. PVT1 and miR-17-5p levels were measured using qRT-PCR analysis. The concentrations of inflammatory cytokines were determined with ELISA kits. Cell viability and apoptosis were assessed using CCK-8 assay and flow-cytometric analysis, respectively. Protein levels were examined using western blot assay. The targeting association between miR-17-5p and PVT1 was verified by dual-luciferase reporter, RIP and RNA pull-down assays. PVT1 level was elevated and miR-17-5p level was declined in septic AKI patients' serum and LPS-stimulated HK-2 cells. Cell viability was suppressed and cell apoptosis and inflammation were promoted after LPS treatment. PVT1 knockdown or miR-17-5p elevation restored LPS-mediated HK-2 cell injury. MiR-17-5p was sponged by PVT1, and its inhibition weakened the impact of PVT1 deficiency on LPS-mediated injury of HK-2 cells. In addition, PVT1 knockdown inactivated NF-κB pathway mediated by LPS treatment, but miR-17-5p inhibition further reversed this effect. PVT1 knockdown promoted cell viability, suppressed inflammatory response and apoptosis by regulating miR-17-5p expression and NF-κB pathway in LPS-stimulated HK-2 cells.

Yuan W ，Xiong X ，Du J ，Fan Q ，Wang R ，Zhang X ... -  《-》