Liu Xinwei, Zhang Jingdong, Han Wenfeng, Wang Yu, Liu Yunen, Zhang Yubiao, Zhou Dapeng, Xiang Liangbi
Department of Orthopaedic Surgery, Rescue Center for Severe Wound and Trauma of Chinese PLA, The General Hospital of Shenyang Military Area Command, Shenyang, Liaoning 110016, P.R. China.
Laboratory of Severe and War‑Related Trauma Center, The General Hospital of Shenyang Military Area Command, Shenyang, Liaoning 110016, P.R. China.
Mol Med Rep. 2017 Jul;16(1):192-200. doi: 10.3892/mmr.2017.6553. Epub 2017 May 9.
The present study aimed to investigate the role of Bruton's tyrosine kinase (BTK) in the pathogenesis of lung injury induced by trauma‑hemorrhagic shock (THS), and to examine the pulmonary protective effects of BTK inhibition. Male Sprague‑Dawley rats were divided into four groups (n=12/group): i) A Sham group, which received surgery without induced trauma; ii) a THS‑induced injury group; iii) a THS‑induced injury group that also received treatment with the BTK inhibitor LFM‑A13 prior to trauma induction; and iv) a Sham group that was pretreated with LFM‑A13 prior to surgery but did not receive induced trauma. The expression of phosphorylated‑BTK protein in the lungs was measured by immunohistochemistry and western blot analysis. The bronchoalveolar lavage fluid (BALF) protein concentration, total leukocyte and eosinophil numbers, and the expression levels of peripheral blood proinflammatory factors were measured. Morphological alterations in the lungs were detected by hematoxylin and eosin staining. Pulmonary nitric oxide (NO) concentration and inducible NO synthase (iNOS) expression were also assessed. Activities of the nuclear factor (NF)‑κB and mitogen‑activated protein kinase (MAPK) signaling pathways were determined by western blotting or electrophoretic mobility shift assay. BTK was notably activated in lungs of THS rats. BALF protein concentration, total leukocytes and eosinophils, peripheral blood expression levels of tumor necrosis factor‑α, interleukin (IL)‑1β, IL‑6 and monocyte chemotactic protein 1 were significantly upregulated after THS induction, and each exhibited decreased expression upon LFM‑A13 treatment. THS‑induced interstitial hyperplasia, edema and neutrophilic infiltration in lungs were improved by the inhibition of BTK. In addition, THS‑induced NO release, iNOS overexpression, and NF‑κB and MAPK signaling were suppressed by BTK inhibition. Results from the present study demonstrate that BTK may serve a pivotal role in the pathogenesis of THS‑related lung injury, and the inhibition of BTK may significantly alleviate THS‑induced lung damage. These results provide a potential therapeutic application for the treatment of THS‑induced lung injury.
本研究旨在探讨布鲁顿酪氨酸激酶（BTK）在创伤失血性休克（THS）诱导的肺损伤发病机制中的作用，并研究抑制BTK的肺保护作用。将雄性Sprague-Dawley大鼠分为四组（每组n = 12）：i）假手术组，接受未诱导创伤的手术；ii）THS诱导损伤组；iii）在诱导创伤前也接受BTK抑制剂LFM-A13治疗的THS诱导损伤组；iv）在手术前用LFM-A13预处理但未接受诱导创伤的假手术组。通过免疫组织化学和蛋白质印迹分析测定肺中磷酸化BTK蛋白的表达。测量支气管肺泡灌洗液（BALF）蛋白浓度、总白细胞和嗜酸性粒细胞数量以及外周血促炎因子的表达水平。通过苏木精和伊红染色检测肺中的形态学改变。还评估了肺一氧化氮（NO）浓度和诱导型NO合酶（iNOS）表达。通过蛋白质印迹或电泳迁移率变动分析确定核因子（NF）-κB和丝裂原活化蛋白激酶（MAPK）信号通路的活性。BTK在THS大鼠的肺中显著激活。THS诱导后，BALF蛋白浓度、总白细胞和嗜酸性粒细胞、外周血肿瘤坏死因子-α、白细胞介素（IL）-1β、IL-6和单核细胞趋化蛋白1的表达水平显著上调，而在LFM-A13治疗后各指标表达均降低。抑制BTK可改善THS诱导的肺间质增生、水肿和中性粒细胞浸润。此外，抑制BTK可抑制THS诱导的NO释放、iNOS过表达以及NF-κB和MAPK信号传导。本研究结果表明，BTK可能在THS相关肺损伤的发病机制中起关键作用，抑制BTK可显著减轻THS诱导的肺损伤。这些结果为THS诱导的肺损伤治疗提供了潜在的治疗应用。
