Effect of the HIF-1 α/miR-23a pathway in the renal tubules on sepsis-associated acute kidney injury and the underlying mechanism
Objective To investigate the role of renal tubules hypoxia inducible factor-1α(HIF-1 α)/MicroRNA-23a(miR-23a)pathway in sepsis-associated acute kidney injury(SA-AKI)and its related mechanisms.Methods Human proximal convoluted tubules epithelial cells(HK-2 cells)were cultured in vitro and treated with lipopolysaccharide(LPS)to construct the SA-AKI cell model.LPS-treated HK-2 cells were divided into LPS group,NC siRNA group,HIF-1α siRNA group,anti-miR-NC group,anti-miR-23a group,HIF-1α siRNA+miR-NC group,and HIF-1α siRNA+miR-23a group.HK-2 cells treated with blank control were considered as the control group.Relative expressions of HIF-1α,miR-23a in cells were detected by quantitative reverse transcriptase PCR(qRT-PCR).Cell viability and apoptosis ere detected by CCK-8 assay and flow cytometry,respectively.The levels of inflammatory factors,including interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor α(TNF-α)in cells were detected by enzyme-linked immunosorbent assay(ELISA).Protein expressions of HIF-1α,and those in the nuclear factor kappa B(NF-κB)pathway were detected by Western blot.Results Compared with those of control group,HK-2 cells in LPS group presented significantly higher protein and mRNA levels of HIF-1α,mRNA level of miR-23a,apoptotic rate and relative levels of IL-6,IL-1β and TNF-α,but significantly lower cell viability(P<0.05).Compared with those of LPS group,HK-2 cells in HIF-1α siRNA group presented significantly lower protein and mRNA levels of HIF-1α,mRNA level of miR-23a,apoptotic rate and relative levels of IL-6,IL-1β and TNF-α,but significantly higher cell viability(P<0.05).Compared with those of LPS group,HK-2 cells in anti-miR-23a group presented significantly lower mRNA level of miR-23a,apoptotic rate and relative levels of IL-6,IL-1β and TNF-α,but significantly higher cell viability(P<0.05).Compared with those of HIF-1α siRNA group,HK-2 cells in HIF-1α siRNA + miR-23a group presented significantly higher mRNA level of miR-23a,apoptotic rate and relative levels of IL-6,IL-1β and TNF-α,but significantly lower cell viability(P<0.05).Compared with those of control group,HK-2 cells in LPS group presented significantly higher levels of p-NF-κB p65/NF-κB p65 and p-IκBα/IκBα(P<0.05).Compared with those of LPS group,p-NF-κB p65/NF-κB p65 and p-IκBα/IκBα were significantly lower in the HIF-1α siRNA group and anti-miR-23a group(P<0.05).Compared with those of HIF-1α siRNA group,p-NF-κB p65/NF-κB p65 and p-IκBα/IκBα were significantly higher in the HIF-1α siRNA + miR-23a group(P<0.05).Conclusion Renal tubules HIF-1α regulates the NF-κB signaling pathway by regulating the expression level of miR-23a,thus participating in LPS-induced renal tubular epithelial cell injury.
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