Department of Pediatric, Shandong Provincial Hospital, Shandong University, Jinan, Shandong, China.
Immun Inflamm Dis. 2023 Mar;11(3):e809. doi: 10.1002/iid3.809.
Acute lung injury (ALI) is a severe and fatal respiratory disease. SIRT6 exerts pivotal activities in the process of lung diseases, but whether SIRT6 impacts ALI has not been covered.
Lentivirus recombinant expressing vector SIRT6 gene (Lent-SIRT6) was constructed in mice, and there were control, lipopolysaccharide (LPS), LPS + Vehicle, and LPS + Lent SIRT6 groups. RT-qPCR and western blot detected SIRT6 expression in lung tissues. HE staining observed pathological alternations in lung tissues. Wet-to-dry ratio of the lungs was then measured. The cell count of bronchoalveolar lavage fluid (BALF) was evaluated. Serum inflammation was examined with enzyme-linked immunosorbent assay, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), and western blot were to measure apoptosis. Western blot tested the expression of ACE2/STAT3/PIM1 signaling-associated factors. At the cellular level, LPS was used to induce lung epithelial cells BEAS-2B to establish cell injury models. SIRT6 was overexpressed and ACE2 expression was inhibited by cell transfection, and the mechanism of SIRT6 in LPS-induced lung injury model was further explored by Cell Counting Kit-8 (CCK-8), western blot, quantitative reverse-transcription polymerase chain reaction, TUNEL, and other techniques.
The results of animal experiments showed that SIRT6 overexpression could reduce LPS-induced lung pathological injury, pulmonary edema, and BALF cell ratio and attenuate LPS-induced inflammatory response and cell apoptosis. In the above process, ACE2, STAT3, p-STAT3, and PIM1 expression were affected. In cell experiments, SIRT6 expression was reduced in LPS-induced BEAS-2B cells. Inhibition of ACE2 expression could reverse the inhibitory effect of SIRT6 overexpression on ACE2/STAT3/PIM1 pathway, and cellular inflammatory response and apoptosis.
SIRT6 eased LPS-evoked inflammation and apoptosis of lung epithelial cells in ALI through ACE2/STAT3/PIM1 signaling.
急性肺损伤（ALI）是一种严重且致命的呼吸系统疾病。SIRT6 在肺部疾病的发生发展过程中发挥着关键作用，但 SIRT6 是否影响 ALI 尚未得到证实。
构建 SIRT6 基因慢病毒重组表达载体（Lent-SIRT6），构建小鼠模型，分为对照组、脂多糖（LPS）组、LPS+Vehicle 组和 LPS+Lent-SIRT6 组。采用 RT-qPCR 和 Western blot 检测肺组织中 SIRT6 的表达。HE 染色观察肺组织的病理变化。测量肺组织湿干重比。评估支气管肺泡灌洗液（BALF）细胞计数。酶联免疫吸附试验（ELISA）检测血清炎症因子，末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）和 Western blot 检测细胞凋亡。Western blot 检测 ACE2/STAT3/PIM1 信号相关因子的表达。在细胞水平上，用 LPS 诱导肺上皮细胞 BEAS-2B 建立细胞损伤模型。通过细胞转染过表达 SIRT6，抑制 ACE2 的表达，进一步探讨 SIRT6 在 LPS 诱导的肺损伤模型中的作用机制，采用细胞计数试剂盒（CCK-8）、Western blot、定量逆转录聚合酶链反应（qRT-PCR）、TUNEL 等技术。
动物实验结果表明，SIRT6 过表达可减轻 LPS 诱导的肺组织病理损伤、肺水肿和 BALF 细胞比例，减轻 LPS 诱导的炎症反应和细胞凋亡。在上述过程中，ACE2、STAT3、p-STAT3 和 PIM1 的表达受到影响。在细胞实验中，LPS 诱导的 BEAS-2B 细胞中 SIRT6 表达减少。抑制 ACE2 表达可逆转 SIRT6 过表达对 ACE2/STAT3/PIM1 通路的抑制作用，减轻细胞炎症反应和凋亡。
SIRT6 通过 ACE2/STAT3/PIM1 信号减轻 LPS 诱导的 ALI 肺上皮细胞炎症和凋亡。
