目的 探讨G蛋白通路抑制因子2（G protein Pathway Suppressor 2,GPS2）在机械通气相关肺损伤（Ventilator-induced lung injury，VILI）中的作用机制。方法 人肺微血管内皮细胞（Human pulmonary microvascular endothelial cells，HPMVEC）转染GPS2 siRNA并进行幅度为20%、频率为0.5HZ的循环拉伸的方法建立机械牵张模型。野生型C57BL/6小鼠在机械通气前通过尾静脉注射GPS2 siRNA转染复合物。通过Western Blot法检测黏附连接蛋白p120、紧密连接蛋白Occludin和GPS2的表达，免疫荧光法检测p120和Occludin的定位及表达，流式细胞仪检测活性氧（Reactive Oxygen Species，ROS）和线粒体膜电位（Mitochondrial Membrane Potential，MMP）的含量，H-E染色观察病理学结果，检测肺组织中丙二醛(（Malondialdehyde, MDA）和超氧化物歧化酶（Superoxide dismutase, SOD）水平。结果 敲低GPS2会加重机械牵张导致的p120、Occludin表达降低，进一步加重氧化应激，细胞ROS水平升高、MMP下降，肺组织MDA值升高，SOD活力降低，肺组织出现明显损伤。结论 GPS2可通过调控线粒体功能进而参与机械通气肺损伤。
Objective To investigate the regulating role of G protein Pathway Suppressor 2 (GPS2) in Ventilator-induced lung injury (VILI). Methods Human pulmonary microvascular endothelial cells (HPMVEC) were transfected with GPS2 siRNA and subjected to cyclic stretch with a amplitude of 20% and a frequency of 0.5HZ to establish a mechanical stretch model. Wild-type C57BL/6 mice were injected with the GPS2 siRNA transfection complex via the tail vein before mechanical ventilation. The expression of adhesion junction protein p120, tight junction proteins Occludin and GPS2 were detected by Western Blot. The localization and expression of p120 and Occludin were detected by immunofluorescence. ROS) and Mitochondrial Membrane Potential (MMP), and H-E staining was used to observe the pathological results. The levels of Malondialdehyde (MDA) and Superoxide dismutase (SOD) in lung tissue were detected. Results Knockdown of GPS2 aggravated the reduction of p120 and Occludin expression caused by mechanical stretch, further aggravated oxidative stress, increased ROS level, decreased MMP, increased MDA value, decreased SOD activity in lung tissue, and obvious lung tissue damage. Conclusions GPS2 can be involved in mechanical ventilation lung injury by regulating mitochondrial function.
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