Receptor Interacting Protein 3-Mediated Necroptosis Promotes Lipopolysaccharide-Induced Inflammation and Acute Respiratory Distress Syndrome in Mice.
Receptor Interacting Protein 3-Mediated Necroptosis Promotes Lipopolysaccharide-Induced Inflammation and Acute Respiratory Distress Syndrome in Mice.
复制标题
受体相互作用蛋白 3 介导的坏死性凋亡促进脂多糖诱导的小鼠炎症和急性呼吸窘迫综合征
DOI：
10.1371/journal.pone.0155723
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Wu Q
中科院分区：
文献类型：
--
作者：
Wang L;Wang T;Li H;Liu Q;Zhang Z;Xie W;Feng Y;Socorburam T;Wu G;Xia Z;Wu Q
坏死放大炎症反应，在急性呼吸窘迫综合征（ARDS）中起重要作用。坏死性凋亡是一种新近发现的由受体相互作用蛋白3（RIP 3）介导的程序性坏死。然而，脂多糖（LPS）诱导的ARDS中坏死性凋亡的潜在参与和影响仍然未知。因此，我们探讨了RIP 3介导的坏死性凋亡在LPS诱导的ARDS中的作用和机制。以递增剂量的LPS灌胃诱导小鼠不同程度的ARDS。收获肺组织进行组织学和TUNEL染色，并进行RIP 3、p-RIP 3、X连锁凋亡抑制蛋白（XIAP）、混合谱系激酶结构域样蛋白（MLKL）、总半胱天冬酶-3/8和裂解半胱天冬酶-3/8的蛋白质印迹。然后，用30 mg/kg LPS诱导野生型和RIP 3基因敲除小鼠的ARDS。碘化丙啶（PI）染色标记肺细胞坏死。检测TNF-α、IL-1 β、IL-6、IL-1α、IL-10和HMGB 1水平、组织髓过氧化物酶（MPO）活性、中性粒细胞计数和总蛋白浓度。结果显示，在高剂量LPS（30 mg/kg和40 mg/kg）诱导的重度ARDS中，RIP 3蛋白表达明显增加，同时伴有p-RIP 3和MLKL表达的增加;而在低剂量LPS（10 mg/kg和20 mg/kg）诱导的轻度ARDS中，细胞凋亡明显增加。RIP 3基因敲除可减轻LPS诱导的严重ARDS患者的低温症状，提高患者的存活率，减轻肺组织损伤。RIP 3基因敲除可减轻LPS诱导的IL-1α/β、IL-6和HMGB 1释放增加，降低组织MPO活性，减少BALF中中性粒细胞内流和总蛋白浓度。此外，在LPS诱导的ARDS中，RIP 3耗竭减少了肺中的坏死细胞并降低了MLKL的表达，但对裂解的半胱天冬酶-3没有影响。结论：RIP 3介导的坏死性凋亡是高剂量LPS诱导的小鼠严重ARDS中炎症增强和肺组织损伤的主要机制。
Necrosis amplifies inflammation and plays important roles in acute respiratory distress syndrome (ARDS). Necroptosis is a newly identified programmed necrosis that is mediated by receptor interacting protein 3 (RIP3). However, the potential involvement and impact of necroptosis in lipopolysaccharide (LPS)-induced ARDS remains unknown. We therefore explored the role and mechanism of RIP3-mediated necroptosis in LPS-induced ARDS. Mice were instilled with increasing doses of LPS intratracheally to induce different degrees of ARDS. Lung tissues were harvested for histological and TUNEL staining and western blot for RIP3, p-RIP3, X-linked inhibitor of apoptosis protein (XIAP), mixed lineage kinase domain-like protein (MLKL), total and cleaved caspases-3/8. Then, wild-type and RIP3 knock-out mice were induced ARDS with 30 mg/kg LPS. Pulmonary cellular necrosis was labeled by the propidium Iodide (PI) staining. Levels of TNF-a, Interleukin (IL)-1β, IL-6, IL-1α, IL-10 and HMGB1, tissue myeloperoxidase (MPO) activity, neutrophil counts and total protein concentration were measured. Results showed that in high dose LPS (30mg/kg and 40mg/kg) -induced severe ARDS, RIP3 protein was increased significantly, accompanied by increases of p-RIP3 and MLKL, while in low dose LPS (10mg/kg and 20mg/kg) -induced mild ARDS, apoptosis was remarkably increased. In LPS-induced severe ARDS, RIP3 knock-out alleviated the hypothermia symptom, increased survival rate and ameliorated the lung tissue injury RIP3 depletion also attenuated LPS-induced increase in IL-1α/β, IL-6 and HMGB1 release, decreased tissue MPO activity, and reduced neutrophil influx and total protein concentration in BALF in severe ARDS. Further, RIP3 depletion reduced the necrotic cells in the lung and decreased the expression of MLKL, but had no impact on cleaved caspase-3 in LPS-induced ARDS. It is concluded that RIP3-mediated necroptosis is a major mechanism of enhanced inflammation and lung tissue injury in high dose LPS- induced severe ARDS in mice.