摘要:
Background::MicroRNAs are closely associated with the progression and outcomes of multiple human diseases, including sepsis. In this study, we examined the role of miR-23a in septic injury.Methods:Lipopolysaccharide (LPS) was used to induce sepsis in a rat model and H9C2 and HK-2 cells. miR-23a expression was evaluated in rat myocardial and kidney tissues, as well as H9C2 and HK-2 cells. A miR-23a mimic was introduced into cells to identify the role of miR-23a in cell viability, apoptosis, and the secretion of inflammatory cytokines. Furthermore, the effect of Rho-associated kinase 1 (
ROCK1), a miR-23a target, on cell damage was evaluated, and molecules involved in the underlying mechanism were identified.
Results::In the rat model, miR-23a was poorly expressed in myocardial (sham
vs. sepsis 1.00 ± 0.06
vs. 0.27 ± 0.03,
P < 0.01) and kidney tissues (sham
vs. sepsis 0.27 ± 0.03
vs. 1.00 ± 0.06,
P < 0.01). Artificial overexpression of miR-23a resulted in increased proliferative activity (DNA replication rate: Control
vs. LPS
vs. LPS + Mock
vs. LPS + miR-23a: H9C2 cells: 34.13 ± 3.12
vs. 12.94 ± 1.21
vs. 13.31 ± 1.43
vs. 22.94 ± 2.26,
P < 0.05; HK-2 cells: 15.17 ± 1.43
vs. 34.52 ± 3.46
vs. 35.19 ± 3.12
vs. 19.87 ± 1.52,
P < 0.05), decreased cell apoptosis (Control
vs. LPS
vs. LPS + Mock
vs. LPS + miR-23a: H9C2 cells: 11.39 ± 1.04
vs. 32.57 ± 2.29
vs. 33.08 ± 3.12
vs. 21.63 ± 2.35,
P < 0.05; HK-2 cells: 15.17 ± 1.43
vs. 34.52 ± 3.46
vs. 35.19 ± 3.12
vs. 19.87 ± 1.52,
P < 0.05), and decreased production of inflammatory cytokines, including interleukin-6 (Control
vs. LPS
vs. LPS + Mock
vs. LPS + miR-23a: H9C2 cells: 59.61 ± 5.14
vs. 113.54 ± 12.30
vs. 116.51 ± 10.69
vs. 87.69 ± 2.97 ng/mL;
P < 0.05,
F = 12.67, HK-2 cells: 68.12 ± 6.44
vs. 139.65 ± 16.62
vs. 143.51 ± 13.64
vs. 100.82 ± 9.74 ng/mL,
P < 0.05,
F = 9.83) and tumor necrosis factor-α (Control
vs. LPS
vs. LPS + Mock
vs. LPS + miR-23a: H9C2 cells: 103.20 ± 10.31
vs. 169.67 ± 18.84
vs. 173.61 ± 15.91
vs. 133.36 ± 12.32 ng/mL,
P < 0.05,
F = 12.67, HK-2 cells: 132.51 ± 13.37
vs. 187.47 ± 16.74
vs. 143.51 ± 13.64
vs. 155.79 ± 15.31 ng/mL,
P < 0.05,
F = 9.83) in cells. However,
ROCK1 was identified as a miR-23a target, and further up-regulation of
ROCK1 mitigated the protective function of miR-23a in LPS-treated H9C2 and HK-2 cells. Moreover,
ROCK1 suppressed sirtuin-1 (
SIRT1) expression to promote the phosphorylation of nuclear factor-kappa B (NF-κB) p65, indicating the possible involvement of this signaling pathway in miR-23a-mediated events.
Conclusion::Our results indicate that miR-23a could suppress LPS-induced cell damage and inflammatory cytokine secretion by binding to
ROCK1, mediated through the potential participation of the
SIRT1/NF-κB signaling pathway.