Department of Oncology, Chongqing Traditional Chinese Medicine Hospital, 400021, Chongqing, China.
J Radiat Res. 2024 May 23;65(3):291-302. doi: 10.1093/jrr/rrae017.
This study was aimed to investigate the effect of hydrogen-rich solution (HRS) on acute radiation pneumonitis (ARP) in rats. The ARP model was induced by X-ray irradiation. Histopathological changes were assessed using HE and Masson stains. Inflammatory cytokines were detected by ELISA. Immunohistochemistry and flow cytometry were performed to quantify macrophage (CD68) levels and the M2/M1 ratio. Western blot analysis, RT-qPCR, ELISA and flow cytometry were used to evaluate mitochondrial oxidative stress injury indicators. Immunofluorescence double staining was performed to colocalize CD68/LC3B and p-AMPK-α/CD68. The relative expression of proteins associated with autophagy activation and the adenosine 5'-monophosphate-activated protein kinase/mammalian target of rapamycin/Unc-51-like kinase 1 (AMPK/mTOR/ULK1) signaling pathway were detected by western blotting. ARP decreased body weight, increased the lung coefficient, collagen deposition and macrophage infiltration and promoted M1 polarization in rats. After HRS treatment, pathological damage was alleviated, and M1 polarization was inhibited. Furthermore, HRS treatment reversed the ARP-induced high levels of mitochondrial oxidative stress injury and autophagy inhibition. Importantly, the phosphorylation of AMPK-α was inhibited, the phosphorylation of mTOR and ULK1 was activated in ARP rats and this effect was reversed by HRS treatment. HRS inhibited M1 polarization and alleviated oxidative stress to activate autophagy in ARP rats by regulating the AMPK/mTOR/ULK1 signaling pathway.
本研究旨在探讨富氢溶液（HRS）对大鼠急性放射性肺炎（ARP）的影响。采用 X 射线照射诱导 ARP 模型。使用 HE 和 Masson 染色评估组织病理学变化。通过 ELISA 检测炎症细胞因子。通过免疫组化和流式细胞术定量检测巨噬细胞（CD68）水平和 M2/M1 比值。通过 Western blot 分析、RT-qPCR、ELISA 和流式细胞术评估线粒体氧化应激损伤指标。通过免疫荧光双重染色共定位 CD68/LC3B 和 p-AMPK-α/CD68。通过 Western blot 检测与自噬激活相关的蛋白的相对表达以及腺苷 5'-单磷酸激活蛋白激酶/哺乳动物雷帕霉素靶蛋白/UNC-51 样激酶 1（AMPK/mTOR/ULK1）信号通路。ARP 降低了大鼠的体重，增加了肺系数、胶原沉积和巨噬细胞浸润，并促进了 M1 极化。HRS 治疗后，病理损伤减轻，M1 极化受到抑制。此外，HRS 治疗逆转了 ARP 诱导的线粒体氧化应激损伤和自噬抑制的高表达。重要的是，AMPK-α 的磷酸化在 ARP 大鼠中受到抑制，mTOR 和 ULK1 的磷酸化被激活，而 HRS 治疗则逆转了这种效应。HRS 通过调节 AMPK/mTOR/ULK1 信号通路抑制 M1 极化和缓解氧化应激来激活 ARP 大鼠的自噬。
