Emergency Medicine, Department of General Hospital of Northern Theater Command, Laboratory of Rescue Center of Severe Trauma PLA, No. 83 Road, Shenhe District, Shenyang l10016, China.
Radiation Oncology, Department of General Hospital of Northern Theater Command, Shenyang l10016, China.
Oxid Med Cell Longev. 2019 Sep 2;2019:4848560. doi: 10.1155/2019/4848560. eCollection 2019.
Although CD28 is associated with the expression of inflammatory mediators, apoptosis-related protein, immunosuppression, and tumorigenesis, the effects of CD28 deficiency on blast exposure-induced lung injury have not been investigated. In this study, we have explored the effects of CD28 on blast exposure-induced lung injury and studied its potential molecular mechanisms. A mouse model of blast exposure-induced acute lung injury was established. Sixty C57BL/6 wild-type (WT) and CD28 knockout (CD28) mice were randomly divided into control or model groups. Lung tissue samples were collected 24 h and 48 h after blast injury. Histopathological changes and the expressions of inflammatory-related proteins were detected by hematoxylin-eosin, immunohistochemistry, and immunofluorescence staining. Apoptosis and oxidative stress were evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining and reactive oxygen species (ROS). Inflammation, apoptosis, oxidative stress, and related pathway protein expression were studied by western blotting. In addition, the levels of CD3 and CD28 proteins were measured by flow cytometry. In the current study, we found that CD28 deficiency significantly inhibited blast exposure-induced increases in the lung weight/body weight ratio and wet weight/dry weight ratio; decreased the infiltration of CD44 leukocytes, CD163 macrophages, and CD3 T cells into the lungs; reduced the expressions of proinflammatory cytokines including IL-1, TNF-, and IL-6; and markedly increased IL-10 expression. CD28 deficiency also significantly attenuated blast exposure-induced ROS, MDA5, and IRE expressions; increased SOD-1 expression; lowered the number of apoptotic cells and Bax, Caspase-3, and active Caspase-8 expressions; and increased Bcl-2 expression. Additionally, CD28 deficiency significantly ameliorated blast exposure-induced increases of p-PI3K and p-Akt and ameliorated the decrease in the p-FoxO1 expression. Our results suggest that CD28 deficiency has a protective effect on blast exposure-induced lung injury, which might be associated with the PI3K/Akt/FoxO1 signaling pathway.
虽然 CD28 与炎症介质、凋亡相关蛋白、免疫抑制和肿瘤发生有关，但 CD28 缺乏对爆震伤诱导的肺损伤的影响尚未得到研究。在这项研究中，我们探讨了 CD28 对爆震伤诱导的肺损伤的影响，并研究了其潜在的分子机制。建立了爆震伤诱导的急性肺损伤小鼠模型。将 60 只 C57BL/6 野生型（WT）和 CD28 敲除（CD28）小鼠随机分为对照组和模型组。在爆震伤后 24 小时和 48 小时采集肺组织样本。通过苏木精-伊红、免疫组织化学和免疫荧光染色检测组织病理学变化和炎症相关蛋白的表达。通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）染色和活性氧（ROS）评估细胞凋亡和氧化应激。通过 Western blot 研究炎症、凋亡、氧化应激和相关通路蛋白的表达。此外，通过流式细胞术测量 CD3 和 CD28 蛋白的水平。在本研究中，我们发现 CD28 缺乏显著抑制了爆震伤诱导的肺重/体重比和湿重/干重比的增加；减少了 CD44 白细胞、CD163 巨噬细胞和 CD3 T 细胞向肺部的浸润；降低了包括白细胞介素 1（IL-1）、肿瘤坏死因子-α（TNF-α）和白细胞介素 6（IL-6）在内的促炎细胞因子的表达；并显著增加了白细胞介素 10（IL-10）的表达。CD28 缺乏还显著抑制了爆震伤诱导的 ROS、MDA5 和 IRE 的表达；增加了 SOD-1 的表达；降低了凋亡细胞的数量和 Bax、Caspase-3 和活性 Caspase-8 的表达；并增加了 Bcl-2 的表达。此外，CD28 缺乏显著改善了爆震伤诱导的 p-PI3K 和 p-Akt 的增加，并改善了 p-FoxO1 表达的降低。我们的结果表明，CD28 缺乏对爆震伤诱导的肺损伤具有保护作用，这可能与 PI3K/Akt/FoxO1 信号通路有关。
