摘要目的:探讨长链非编码RNA（lncRNA）-AC013472.3对脂多糖（LPS）刺激大鼠肺泡巨噬细胞（NR8383细胞）分泌肿瘤坏死因子（TNF）-α的影响。方法:以NR8383细胞分别建立lncRNA-AC013472.3沉默及过表达模型，沉默模型分为乱序无意义阴性小干扰RNA序列（si-con）组（转染si-con的NR8383细胞）、LPS+si-con组（10 μg/L的LPS处理转染si-con的NR8383细胞24 h）、小干扰RNA（siRNA）组（转染siRNA的NR8383细胞）和LPS+siRNA组（10 μg/L的LPS处理转染siRNA的NR8383细胞24 h）；过表达模型分为空质粒（p-con）组（转染p-con的NR8383细胞）、LPS+p-con组（10 μg/L的LPS处理转染p-con的NR8383细胞24 h）、lncRNA过表达质粒（plncRNA）组（转染plncRNA的NR8383细胞）和LPS+plncRNA组（10 μg/L的LPS处理转染plncRNA的NR8383细胞24 h），实时荧光定量PCR（qPCR）方法检测各组细胞中TNF-α mRNA相对表达量；Western印迹法检测TNF受体相关因子-6（TRAF-6）和磷酸化的核因子-κB（NF-κB）p65蛋白相对表达量。结果:在沉默模型中，LPS+si-con组TNF-α mRNA相对表达量、TRAF-6和NF-κB p65蛋白相对表达量均显著高于si-con组（2.040±0.195比1.048±0.207、0.473±0.022比0.293±0.076和0.469±0.062比0.252±0.038）（均 P<0.05）；LPS+siRNA组TNF-α mRNA相对表达量、TRAF-6和NF-κB p65蛋白相对表达量均显著高于siRNA组（4.158±0.119比1.028±0.019、0.700±0.104比0.231±0.023和0.771±0.095比0.258±0.050）（均 P<0.05）；LPS+siRNA组各指标相对表达量显著高于LPS+si-con组（均 P<0.05）。在过表达模型中，LPS+p-con组TNF-α mRNA相对表达量、TRAF-6和NF-κB p65蛋白相对表达量均显著高于p-con组（1.961±0.169比0.999±0.143、0.533±0.047比0.247±0.020和0.565±0.108比0.276±0.048）（均 P<0.05）；LPS+plncRNA组TNF-α mRNA、TRAF-6和NF-κB p65蛋白相对表达量均显著高于plncRNA组（1.322±0.110比1.043±0.093、0.347±0.035比0.232±0.023和0.405±0.072比0.268±0.031）（均 P<0.05）；LPS+plncRNA组各指标相对表达量显著低于LPS+p-con组（均 P<0.05）。 结论:lncRNA-AC013472.3可能通过抑制NF-κB信号通路的激活，从而抑制LPS刺激NR8383细胞分泌TNF-α。

abstractsObjective:To investigate the effect of long non-coding RNA-AC013472.3 on lipopolysaccharide (LPS)-stimulated secretion of tumor necrosis factor (TNF)-α in NR8383 rat alveolar macrophages.Methods:Silencing and overexpression models of lncRNA-AC013472.3 were established with NR8383 rat alveolar macrophages as the experimental subjects. The silencing models were divided into three groups: random nonsense negative small interfering RNA sequence (si-con) group (si-con group, si-con transfected NR8383 cells), LPS+si-con group (10 μg/L LPS was used to treat si-con transfected NR8383 cells for 24 h), and siRNA group (siRNA transfected NR8383 cells), and LPS+siRNA group (10 μg/L LPS was used to treat siRNA transfected NR8383 cells for 24 h). The overexpression models were divided into the empty plasmid (p-con) group (p-con transfected NR8383 cells), LPS+p-con group (10 μg/L LPS was used to treat p-con transfected NR8383 cells for 24 h), lncRNA overexpression plasmid (plncRNA) group (plncRNA transfected NR8383 cells), and the LPS+plncRNA group (10 μg/L LPS was used to treat plncRNA transfected NR8383 cells for 24 h). The mRNA levels of TNF-α in each group were examined by quantitative real-time PCR (qPCR). The protein levels of tumor necrosis factor receptor-related factor-6 (TRAF-6) and phosphorylated nuclear factor-κB (NF-κB) p65 were examined by Western blot.Results:In the silencing model, the mRNA levels of TNF-α, the protein levels of TRAF-6 and NF-κB p65 in the LPS+si-con group were significantly higher than those in the si-con group (2.040±0.195 vs 1.048±0.207, 0.473±0.022 vs 0.293±0.076 and 0.469±0.062 vs 0.252±0.038)(all P<0.05). The mRNA levels of TNF-α, the protein levels of TRAF-6 and NF-κB p65 in the LPS+siRNA group were significantly higher than those in the siRNA group (4.158±0.119 vs 1.028±0.019, 0.700±0.104 vs 0.231±0.023 and 0.771±0.095 vs 0.258±0.050)(all P<0.05). The relative expression levels of all indexes in the LPS+siRNA group were significantly higher than those in the LPS+si-con group (all P<0.05). In the overexpression model, the mRNA levels of TNF-α, the protein levels of TRAF-6 and NF-κB p65 in the LPS+p-con group were significantly higher than those in the p-con group (1.961±0.169 vs 0.999±0.143, 0.533±0.047 vs 0.247±0.020 and 0.565±0.108 vs 0.276±0.048) (all P<0.05). The mRNA levels of TNF-α, the protein levels of TRAF-6 and NF-κB p65 in the LPS+plncRNA group were significantly higher than those in the plncRNA group (1.322±0.110 vs 1.043±0.093, 0.347±0.035 vs 0.232±0.023 and 0.405±0.072 vs 0.268±0.031) (all P<0.05). The relative expression of all indexes in the LPS+plncRNA group were significantly lower than that in the LPS+p-con group (all P<0.05). Conclusion:LncRNA-AC013472.3 may inhibit the activation of NF-κB signaling pathway, thereby inhibiting the LPS-stimulated secretion of TNF-α in NR8383 rat alveolar macrophages.