Department of Orthopaedics, the Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi, China.
Hou Zonglian Medical Experimental Class, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi, China.
J Recept Signal Transduct Res. 2020 Jun;40(3):209-217. doi: 10.1080/10799893.2020.1727925. Epub 2020 Feb 21.
This study aims to investigate the protective effects and possible mechanism of methane-rich saline (MS) on lung ischemia-reperfusion injury (LIRI) in rats. MS (2 ml/kg and 20 ml/kg) was injected intraperitoneally in rats after LIRI. Lung injury was assayed by Hematoxylin-eosin (HE) staining and wet-to-dry weight (W/D). The cells in the bronchoalveolar lavage fluid (BALF) and blood were counted. Oxidative stress was examined by the level of malondialdehyde (MDA) and superoxide dismutase (SOD). Inflammatory factors including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-10 (IL-10) were determined by ELISA. Lung tissue apoptosis was detected by TUNEL staining and western blotting of Bcl-2, Bax, and caspase-3. The expressions of IкBα, p38, PI3K, AKT, and NF-κB were analyzed with Western blotting. MS effectively decreased the lung W/D ratio as well as the lung pathological damage and reduced the localized infiltration of inflammatory cells. Methane suppressed the expression of the PI3K-AKT-NFκB signaling pathway during the lung IR injury, which inhibited the activation of NF-kB and decreased the level of inflammatory cytokines, such as TNF-α, IL-1β, and IL-10. Moreover, we found that MS treatment relieved reactive oxygen species (ROS) damage by downregulating MDA and upregulating SOD. MS treatment also regulated apoptosis-related proteins, such as Bcl-2, Bax, and caspase-3. MS could repair LIRI and reduce the release of oxidative stress, inflammatory cytokines, and cell apoptosis via the PI3K-AKT-NFκB signaling pathway, which may provide a novel and promising strategy for the treatment of LIRI.
本研究旨在探讨富含甲烷的盐水（MS）对大鼠肺缺血再灌注损伤（LIRI）的保护作用及可能机制。在 LIRI 后，大鼠腹腔内注射 MS（2ml/kg 和 20ml/kg）。通过苏木精-伊红（HE）染色和湿重/干重（W/D）比评估肺损伤。计数支气管肺泡灌洗液（BALF）和血液中的细胞。通过丙二醛（MDA）和超氧化物歧化酶（SOD）水平检测氧化应激。通过酶联免疫吸附试验（ELISA）测定肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-10（IL-10）等炎症因子。通过 TUNEL 染色和 Bcl-2、Bax 和 caspase-3 的 Western blot 检测肺组织凋亡。用 Western blot 分析 IκBα、p38、PI3K、AKT 和 NF-κB 的表达。MS 可有效降低肺 W/D 比值及肺组织病理损伤，减少炎症细胞局部浸润。甲烷抑制了肺 IR 损伤过程中 PI3K-AKT-NFκB 信号通路的表达，抑制了 NF-κB 的激活，降低了 TNF-α、IL-1β和 IL-10 等炎症细胞因子的水平。此外，我们发现 MS 处理通过下调 MDA 和上调 SOD 来缓解活性氧（ROS）损伤。MS 处理还调节了凋亡相关蛋白，如 Bcl-2、Bax 和 caspase-3。MS 可通过 PI3K-AKT-NFκB 信号通路修复 LIRI，减少氧化应激、炎症细胞因子和细胞凋亡的释放，这可能为 LIRI 的治疗提供一种新的有前途的策略。
