Intensive Care Unit, Shanghai Eighth People's Hospital, Shanghai 200235, P.R. China.
Mol Med Rep. 2024 Sep;30(3). doi: 10.3892/mmr.2024.13295. Epub 2024 Jul 26.
Sepsis is a life‑threatening multiple organ failure disease caused by an uncontrolled inflammatory response and can progress to acute lung injury (ALI). Heat‑shock protein B8 (HSPB8) serves a cytoprotective role in multiple types of diseases; however, to the best of our knowledge, the regulatory role of HSPB8 in sepsis‑induced ALI remains unclear. A549 human alveolar type II epithelial cells were treated with lipopolysaccharide (LPS) for 24 h to simulate a sepsis‑induced ALI model. Cell transfection was performed to overexpress HSPB8, and cells were treated with mitochondrial division inhibitor‑1 (Mdivi‑1) for 2 h before LPS induction to assess the underlying mechanism. Protein expression was evaluated using western blotting and an immunofluorescence assay. Cytokines were examined using ELISA assay kits and antioxidant enzymes were examined using their detection kits. Cell apoptosis was detected using flow cytometry. The mitochondrial membrane potential was detected by JC‑1 staining. HSPB8 was upregulated in A549 cells treated with LPS and HSPB8 overexpression attenuated LPS‑induced inflammatory cytokine levels, oxidative stress and apoptosis in A549 cells. LPS inhibited mitophagy and reduced the mitochondrial membrane potential in A549 cells, which was partly inhibited by HSPB8 overexpression. Furthermore, Mdivi‑1 decreased the inhibitory effect of HSPB8 on the inflammatory response, oxidative stress and apoptosis in LPS‑treated A549 cells. In conclusion, HSPB8 overexpression attenuated the LPS‑mediated inflammatory response, oxidative stress and apoptosis in A549 cells by promoting mitophagy, indicating HSPB8 as a potential therapeutic target in sepsis‑induced ALI.
脓毒症是一种危及生命的多器官功能衰竭疾病，由失控的炎症反应引起，并可进展为急性肺损伤（ALI）。热休克蛋白 B8（HSPB8）在多种疾病中发挥细胞保护作用；然而，据我们所知，HSPB8 在脓毒症诱导的 ALI 中的调节作用尚不清楚。用脂多糖（LPS）处理 A549 人肺泡 II 型上皮细胞 24 h 模拟脓毒症诱导的 ALI 模型。进行细胞转染以过表达 HSPB8，并在 LPS 诱导前用线粒体分裂抑制剂-1（Mdivi-1）处理 2 h，以评估潜在机制。使用 Western blot 和免疫荧光测定评估蛋白质表达。使用 ELISA 试剂盒检测细胞因子，使用其检测试剂盒检测抗氧化酶。使用流式细胞术检测细胞凋亡。通过 JC-1 染色检测线粒体膜电位。LPS 处理的 A549 细胞中 HSPB8 上调，过表达 HSPB8 可减轻 LPS 诱导的 A549 细胞炎症细胞因子水平、氧化应激和细胞凋亡。LPS 抑制线粒体自噬并降低 A549 细胞的线粒体膜电位，而过表达 HSPB8 可部分抑制该作用。此外，Mdivi-1 降低 HSPB8 对 LPS 处理的 A549 细胞炎症反应、氧化应激和细胞凋亡的抑制作用。综上所述，HSPB8 过表达通过促进线粒体自噬减轻 LPS 介导的 A549 细胞炎症反应、氧化应激和细胞凋亡，表明 HSPB8 可能是脓毒症诱导的 ALI 的潜在治疗靶点。
