RESUMO
MiR-429 was reported to be downregulated in contrast-induced acute kidney injury (CI-AKI). However, whether miR-429 is functionally relevant with CI-AKI needs further investigation. Human renal tubular epithelial cell (HK-2) cells were stimulated with contrast media iodixanol to establish in vitro CI-AKI model. Cell Counting Kit-8 (CCK-8) was applied to access cell viability. Flow cytometry was performed to determine apoptosis. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to evaluate level of programmed cell death 4 (PDCD4) mRNA and miR-429 while western blot was applied to evaluate level of proteins including PDCD4, B-cell leukaemia/lymphoma 2 (Bcl-2), BCL2-associated X protein (Bax), cleaved caspase 3, cleaved caspase 9, p65, phosphorylated p65. Dual luciferase assay was used to validate miR-429 targeting PDCD4. MiR-429 was downregulated whereas PDCD4 was upregulated in contrast media iodixanol-stimulated HK-2 cells. MiR-429 overexpression elevated cell viability and attenuated cell apoptosis. Moreover, the activation of nuclear factor kappa-B (NF-κB) signalling was suppressed after miR-429 overexpression, while PDCD4 overexpression reversed these effects. MiR-429 directly targeted PDCD4 and negatively regulated its expression. CI-AKI induced NF-κB signalling activation and PDCD4 overexpression further promoted NF-κB signalling activation. However, the treatment of BAY11-7082 reversed above results. Overexpression of miR-429 attenuated apoptosis and elevated cell viability in a CI-AKI cell model via targeting PDCD4 and thus restraining NF-κB signalling.
Assuntos
Injúria Renal Aguda , Proteínas Reguladoras de Apoptose , MicroRNAs , Proteínas de Ligação a RNA , Injúria Renal Aguda/induzido quimicamente , Injúria Renal Aguda/genética , Apoptose/genética , Proteínas Reguladoras de Apoptose/genética , Linhagem Celular , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , NF-kappa B/metabolismo , Proteínas de Ligação a RNA/genética , Transdução de SinaisRESUMO
A new and simple colorimetric sensor with high selectivity for acetate ion based on 1,10-phenanthroline-2,9-dicarboxyaldehyde-di-(p-substitutedphenylhydrazone) receptor 2 has been synthesized. The selectively binding ability of receptor 2 to acetate ion over other anions tested was demonstrated by UV-vis absorption spectroscopy in DMSO. Comparing with other anions studied, the UV-vis absorption spectrum in dimethyl sulfoxide shows significant response toward acetate ion with high selectivity, and meanwhile dramatic color change is observed from yellow to green in the presence of acetate ion (5 x 10(-6)mol/L). Little UV-vis absorption spectrum change has occurred when receptor 2 was titrated with other different guest (F(-), Cl(-), Br(-), I(-), H(2)PO(4)(-) and OH(-)). In addition, the (1)H NMR spectrum titration shows deprotonation of the receptor in the presence of acetate ion.
