抑制SUMO特异性蛋白酶1在小鼠急性肺损伤中的保护作用

Protective Effect Associated with Inhibiting SUMO Specific Protease 1 for Acute Lung Injury in Mice

目的:探讨抑制SUMO特异性蛋白酶1/低氧诱导因子-1α(SUMO-specific protease 1/hypoxia-inducible factor-1α,SENP1/HIF-1α)信号通路在减轻脂多糖(lipopolysaccharide,LPS)致小鼠急性肺损伤中的作用及机制。方法:建立小鼠LPS致急性肺损伤模型,采用实时荧光定量聚合酶链反应(Real-Time polymerase chain reaction,Real-Time PCR)法检测小鼠经LPS处理后肺组织中SENP1 mRNA的表达,采用蛋白质印迹(Western印迹)法检测HIF-1α蛋白的表达。采用小干扰RNA法敲除小鼠淋巴瘤巨噬细胞(RAW264.7)SENP1的表达,用LPS处理对照组和干扰组细胞后,采用Western印迹法检测细胞中caspase-3、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的表达,采用酶联免疫吸附试验法检测细胞中髓过氧化物酶(myeloperoxidase,MPO)的活性。结果:小鼠急性肺损伤后肺组织中HIF-1α蛋白和SENP1 mRNA的表达水平均升高。经LPS处理后,干扰组caspase-3、iNOS和MPO较对照组均显著降低(P<0.05)。结论:抑制SENP1/HIF-1α通路可减轻LPS致小鼠急性肺损伤。

Objective ：To explore the effect and mechanism of inhibiting SUMO-specific protease 1/hypoxia-inducible factor-1α（SENP1/HIF-1α） pathway for attenuating acute lung injury induced by lipopolysaccharide （LPS ） in mice .Methods ： LPS induced acute lung injury model in mice was established .The expression of SENP1 mRNA in lung tissue of mice treated by LPS was tested by Real-Time polymerase chain reaction （Real-Time PCR） ,while the expression of HIF-1α protein was tested by Western blotting .Small interfering RNA （siRNA） was used to knock out the expression of SENP1 of mouse leukemic monocyte macrophage cell （RAW 264 .7） .Once the cells from both control group （Ns group） and interfering group （Si group） were treated by LPS ,the expression of caspase-3 ,inducible nitric oxide synthase（iNOS） was detected by Western blotting and the activity of myeloperoxidase（MPO ） was tested by PCR .Results ： The expression levels of both HIF-1α protein and SENP1 mRNA increased in lung tissue of mice after acute lung injury .The levels of caspase-3 ,iNOS and M PO significantly decreased in Si group after the treatment by LPS ,compared with that in Ns group （P〈 0 .05） .Conclusions ：Inhibition of SENP1/HIF-1αpathway may attenuate acute lung injury induced by LPS in mice .