DIVERGENT EFFECTS OF NEUTROPHILS ON FAS-INDUCED PULMONARY INFLAMMATION, APOPTOSIS, AND LUNG DAMAGE
DIVERGENT EFFECTS OF NEUTROPHILS ON FAS-INDUCED PULMONARY INFLAMMATION, APOPTOSIS, AND LUNG DAMAGE
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DOI：
10.1097/shk.0000000000000685
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发表时间：
2017-02-01
期刊：
影响因子：
3.1
通讯作者：
Perl, Mario
中科院分区：
文献类型：
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作者：
Bruns, Bastian;Hoenle, Theresia;Perl, Mario
肺Fas活化在急性呼吸窘迫综合征的发病机制中是必不可少的。Fas诱导的肺损伤是否依赖于中性粒细胞或主要由上皮细胞凋亡引发尚不清楚。肺上皮细胞（LEC）和肺泡巨噬细胞（AM）的贡献仍然难以捉摸。在腹膜内滴注Fas激活（Jo 2）或同种型抗体6或18小时之前，使小鼠中性粒细胞减少。LEC和AM与Jo 2孵育，并在核因子κ B、p-38丝裂原活化蛋白激酶（p38 MAPK）或细胞外信号调节激酶1/2（ERK 1/2）抑制剂存在下孵育。通过流式细胞仪珠阵列或ELISA评估细胞因子。通过活性半胱天冬酶-3蛋白质印迹和末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）定量细胞凋亡。通过支气管肺泡灌洗液（BALF）蛋白浓度和肺组织学评估肺损伤。KC，IL-6和MCP-1显着增加，在肺，血浆和BALF 18小时后，乔在中性粒细胞的存在下，在嗜中性粒细胞减少小鼠肺，MCP-1，但不是KC或IL-6，甚至进一步增强。6小时后，乔2，BALF蛋白显着增加，只有在中性粒细胞的存在。细胞凋亡不受中性粒细胞减少的影响。AM释放MCP-1，并在较低浓度的Jo 2比LEC进行凋亡。抑制p38 MAPK可显著增加AM和LEC的凋亡，而抑制ERK 1/2可减少AM和LEC的凋亡。总之，中性粒细胞是Fas诱导的肺损伤的必要组成部分，而不直接影响肺细胞凋亡本身。LEC对Fas诱导的炎症反应和凋亡的抵抗能力强于AM。
Pulmonary Fas activation is essential in the pathogenesis of the acute respiratory distress syndrome. It remains unclear whether Fas-induced lung injury is dependent on neutrophils or mainly triggered by epithelial cell apoptosis. The contribution of lung epithelial cells (LEC) and alveolar macrophages (AM) remains elusive. Mice were neutrophil reduced prior to intratracheal instillation of Fas-activating (Jo2) or isotype antibody for 6 or 18 h. LEC and AM were incubated with Jo2 and in the presence of nuclear factor kappa B, p-38 mitogen activated protein kinase (p38MAPK), or extracellular signal regulating kinase 1/2 (ERK1/2) inhibitors. Cytokines were assessed by cytometric bead array or ELISA. Apoptosis was quantified via active caspase-3 Western blotting and Terminal Deoxynucleotide Transferase dUTP Nick End Labeling (TUNEL). Lung injury was assessed by bronchoalveolar lavage fluid (BALF) protein concentration and lung histology. KC, IL-6, and MCP-1 were markedly increased in lung, plasma, and BALF 18 h after Jo2 in the presence of neutrophils; in neutrophil-reduced mice lungs, MCP-1, but not KC or IL-6, was even further enhanced. Six hours after Jo2, BALF protein was markedly increased only in the presence of neutrophils. Apoptosis remained unaffected by neutrophil reduction. AM released MCP-1 and underwent apoptosis at lower concentrations of Jo2 than LEC. Inhibition of p38MAPK significantly increased, while inhibition of ERK1/2 reduced AM and LEC apoptosis. In conclusion, neutrophils are a necessary component of Fas-induced lung damage, while not affecting lung apoptosis directly per se. LEC display higher resistance to Fas-triggered inflammation and apoptosis than AM.