Heme oxygenase-1 attenuates seawater drowning-induced acute lung injury through a reduction in inflammation and oxidative stress
Heme oxygenase-1 attenuates seawater drowning-induced acute lung injury through a reduction in inflammation and oxidative stress
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DOI：
10.1016/j.intimp.2019.05.019
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发表时间：
2019-09-01
影响因子：
5.6
通讯作者：
Pang, Qing-feng
中科院分区：
文献类型：
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作者：
Sun, Xue-qian;Wu, Chen;Pang, Qing-feng
目的：血红素氧合酶-1（HO-1）通过其强的抗炎、抗氧化、抗凋亡等特性在各种创伤性急性肺损伤（ALI）中发挥重要的保护作用，但其对海水吸入性急性肺损伤的保护作用及其机制尚不清楚。本研究旨在探讨HO-1对海水吸入性急性肺损伤（ALI）的治疗作用及其机制。方法：采用细胞计数试剂盒-8和乳酸脱氢酶释放法检测A549细胞的存活率和侵袭力;炎症细胞因子的转录水平（TNF-α、IL-6、IL-8和MCP-1）和细胞增殖相关的细胞因子qPCR检测海水处理后A549细胞中FoxMi、Ccnb 1和Cdc 25 C的表达，流式细胞仪检测海水处理后A549细胞凋亡情况;采用qPCR和western blotting检测HO-1 mRNA和蛋白的表达;采用荧光指示剂DCFH-DA、二氢乙锭、MitoSox Red和Fluo-4监测ROS的产生和线粒体功能。采用天狼猩红染色、TUNEL法和抗泛细胞角蛋白抗体免疫组化染色等方法，对海水淹溺小鼠肺组织的湿/干重比值、乳酸脱氢酶活性进行了分析。结果：海水暴露可降低细胞活力，增加促炎细胞因子（IL-6、IL-8和TNF-α）的产生，诱导细胞凋亡，抑制细胞增殖相关因子（FoxM 1、Ccnb 1和Cdc 25 C）的表达。此外，海水暴露导致A549细胞线粒体功能障碍。补充HO-1特异性诱导剂（血红素）或其催化产物（胆绿素）可显著减轻海水诱导的A549细胞损伤，促进细胞增殖。结论：HO-1通过其抗炎、抗氧化和抗细胞凋亡功能减轻海水溺水肺组织损伤。
Objective: Heme oxygenase-1 (HO-1) plays a critical protective role in various insults-induced acute lung injury (ALI) through its strong anti-inflammatory, anti-oxidant, and anti-apoptotic properties, but its protective role and mechanism on seawater aspiration-induced acute lung injury remains unclear. This study aimed to explore the therapeutic potential and mechanism of HO-1 to attenuate seawater aspiration-induced ALI in vivo and in vitro.Methods: The viability and invasion of A549 cell were analyzed through cell counting kit-8 and lactate dehydrogenase release assay; the transcriptional level of inflammatory cytokines (TNF-alpha, IL-6, IL-8 and MCP-1) and cell proliferation-related cytokines (FoxMi, Ccnb1 and Cdc25C) in seawater-treated A549 cell were tested by qPCR; apoptotic cells were analyzed by flow cytometryd; HO-1mRNA and protein were determined by qPCR and western blotting; the fluorescent indicators (DCFH-DA, dihydroethidium, MitoSox Red and Fluo-4) were used to monitor generation of ROS and mitochondrial function. The lung wet/dry weight radio and lactate dehydrogenase activity, Sirius red staining, TUNEL assay and immunohistochemical staining with anti-pan Cytokeratin antibody were analyzed in seawater-drowning mice. The role of HO-1 on seawater-drowning pulmonary injury was explored via HO-1 activity inhibitors (Zinc protoporphyrin) in vitro and in vivo.Results: Seawater exposure decreased the cellular viability, increased the production of pro-inflammatory cytokines (IL-6, IL-8 and TNF-alpha), induced cellular apoptosis and inhibited the expression of cell proliferation-related cytokines (FoxM1, Ccnb1 and Cdc25C). Moreover, seawater exposure led to mitochondrial dysfunction in A549 cells. Supplement of HO-1 sepcific inducer (heme) or its catalytic product (biliverdin) significantly attenuated seawater-induced A549 damage and promoted cell proliferation. However, Zinc protoporphyrin abolished the beneficial effects of HO-1 on seawater drowning-induced pulmonary tissue injury.Conclusion: HO-1 attenuates seawater drowning-induced lung injury by its anti-inflammatory, anti-oxidative, and anti-apoptosis function.