The mechanism by which lncRNA MINCR regulates the AMPK/SIRT1 signaling pathway in LPS-induced human alveolar epithelial cell autophagy and inflammatory response
Objective To explore the mechanism by which lncRNA MINCR regulates the AMPK/SIRT1 sig-naling pathway in LPS-induced autophagy and inflammatory response in human alveolar epithelial cells.Methods A549 cells were randomly divided into the Control group(A549 cells cultured normally),LPS group(A549 cells treated with 10 μg/mL LPS),si-NC group(A549 cells transfected with si-NC plasmid and treated with 10 μg/mL LPS),si-MINCR group(A549 cells transfected with si-MINCR plasmid and treated with 10 μg/mL LPS),and si-MINCR+Compound C group(A549 cells transfected with si-MINCR plasmid and treated with 10 μg/mL LPS and 50μmol/L Compound C).qRT-PCR was used to detect the relative expression of MINCR in cells,CCK-8 assay and flow cytometry were used to assess cell viability and apoptosis rate,ELISA was used to measure the levels of inflam-matory factors IL-6,IL-8,and TNF-α in cells,and Western blotting was used to analyze the protein expression of LC3,p62,Beclin-1,AMPK,p-AMPK,and SIRT1 in cells.Results Compared to the Control group,the LPS group showed a significant increase in the relative expression of MINCR(3.84±0.52),apoptosis rate(26.84％±3.12％),levels of IL-6(416.95±45.06),IL-8(549.32±61.45),TNF-α(694.81±71.42),and the expression of p62 protein(0.87±0.10)in cells,while the ratio of LC3 Ⅱ/LC3Ⅰ(0.32±0.05),p-AMPK/AMPK ratio(0.31±0.05),Beclin-1(0.52±0.07),and SIRT1 protein(0.46±0.06)were significantly decreased(P＜0.01).Compared to the LPS group,the si-MINCR group showed a significant decrease in the relative expression of MINCR(1.45±0.26),apoptosis rate(9.62％±1.15％),levels of IL-6(198.62±21.34),IL-8(287.15±30.76),TNF-α(364.72±40.59),and the expression of p62 protein(0.87±0.10),while the ratio of LC3 Ⅱ/LC3Ⅰ(0.78±0.09),p-AMPK/AMPK ratio(0.64±0.07),Beclin-1(0.73±0.08),and SIRT1 protein(0.81±0.09)were significantly increased(P＜0.01).Com-pared to the si-MINCR group,the si-MINCR+Compound C group showed a significant increase in the relative expres-sion of MINCR(3.21±0.45),apoptosis rate(20.24％±2.26％),levels of IL-6(395.78±10.47),IL-8(543.26±56.71),TNF-α(610.54±65.33),and the expression of p62 protein(0.79±0.08),while the ratio of LC3 Ⅱ/LC3Ⅰ(0.40±0.06),p-AMPK/AMPK ratio(0.38±0.04),Beclin-1(0.56±0.06),and SIRT1 protein(0.52±0.07)were sig-nificantly decreased(P＜0.01).Conclusion Knockdown of lncRNA MINCR promotes LPS-induced autophagy in al-veolar epithelial cells and mitigates inflammatory injury,which may be associated with the activation of the AMPK/SIRT1 signaling pathway.
NETL
NSTL
万方数据
