| 摘要: |
| ［摘要］ 目的 探究还原型谷胱甘肽（GSH）对pristane诱导的系统性红斑狼疮相关弥漫性肺泡出血（SLE-DAH）小鼠模型的保护作用和机制。方法 体内实验：选择6～10周龄C57BL/6J雌性小鼠，腹腔注射0.5 mL pristane构建弥漫性肺泡出血（DAH）模型，其后第3、6、9、12天予腹腔注射GSH，观察小鼠肺组织大体、肺组织病理及体质量变化，检测肺泡灌洗液（BALF）中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）水平。体外实验：β-CD-pristane处理MLE-12细胞，观察细胞形态及密度，评估细胞活力，mRNA-seq分析基因表达变化，GSH干预后检测细胞增殖情况和活性氧（ROS）水平。结果 pristane处理后小鼠肺组织大体可见不同程度出血改变，肺组织大体出血百分比及DAH评分显著升高（P<0.05），肺组织结构紊乱，炎症细胞浸润明显，体质量显著下降（P<0.05），且BALF中TNF-α、IL-6、IL-1β水平显著升高（P<0.05）。β-CD-pristane处理后MLE-12细胞形态改变，活力显著降低（P<0.05），RNA-seq测序结合京都基因与基因组百科全书（KEGG）通路富集分析及基因本体（GO）功能富集分析提示，β-CD-pristane可能通过谷胱甘肽代谢通路与炎症反应致细胞损伤。GSH干预后，MLE-12细胞增殖恢复（P<0.05），细胞内ROS水平显著下降（P<0.05），pristane诱导的DAH小鼠肺组织大体出血百分比及DAH评分显著下降（P<0.05），体质量显著上升（P<0.05），且BALF中TNF-α、IL-6、IL-1β水平显著下降（P<0.05）。结论 GSH干预可减轻pristane诱导的小鼠DAH及炎症反应，其作用机制可能与抑制肺泡上皮细胞氧化应激及促进肺泡上皮细胞增殖有关。 |
| 关键词: 系统性红斑狼疮相关弥漫性肺泡出血 还原型谷胱甘肽 氧化应激 炎症反应 |
| DOI：10.3969/j.issn.1674-3806.2026.02.11 |
| 分类号:R 593.241 |
| 基金项目:重庆市自然科学基金项目（编号：CSTB2024NSCQ-KJFZZDX0049）；重庆市杰出青年科学基金项目（编号：CSTB2023NSCQ-JQX0033） |
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| Study on the protective effect of reduced glutathione on pristane-induced diffuse alveolar hemorrhage in mice |
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LIAN Shaopan1, ZHOU Ying1, LU Conghua2, SHI Chenjun2, LIN Caiyu2, LI Li2, DAI Huanzi1
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1.Department of Rheumatology and Immunology, Army Characteristic Medical Center of PLA, Chongqing 400042, China; 2.Department of Respiratory and Critical Care Medicine, Army Characteristic Medical Center of PLA, Chongqing 400042, China
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| Abstract: |
| ［Abstract］ Objective To explore the protective effect and mechanism of reduced glutathione(GSH) on a mouse model of pristane-induced systemic lupus erythematosus-associated diffuse alveolar hemorrhage(SLE-DAH). Methods In vivo experiments: Female C57BL/6J mice aged 6 to 10 weeks were selected and used to construct a diffuse alveolar hemorrhage(DAH) model by intraperitoneal injection of 0.5 mL pristane. Subsequently, these mice were intraperitoneally injected with GSH on the third day, the 6th day, the 9th day and the 12th day after constructing the model to observe the gross changes in the lungs, pathological changes in lung tissues, and changes in body weight of the mice. The levels of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-1β(IL-1β) in the bronchoalveolar lavage fluid(BALF) of the mice were detected. In vitro experiments: Mouse lung epithelial(MLE)-12 cells were treated with β-CD-pristane and the morphology and density of the cells were observed, and the viability of the cells was assessed. The changes in gene expressions were analyzed by using miRNA-sequencing(mRNA-seq), and the cell proliferation and the level of reactive oxygen species(ROS) were detected after GSH intervention. Results After the treatment with pristane, the lungs of the mice showed the changes in different degrees of hemorrhage on the whole, and the percentage of gross lung hemorrhage in the lung tissues and DAH scores were significantly increased(P<0.05), with disordered lung tissue structure, significant infiltration of inflammatory cells and significantly decreased weigh(P<0.05), and the levels of TNF-α, IL-6, IL-1β in BALF were significantly increased(P<0.05). After the treatment with β-CD-pristane, the morphology of MLE-12 cells changed, and their viability was significantly decreased(P<0.05). The combination of RNA sequencing(RNA-seq) with Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis and Gene Ontology(GO) functional enrichment analysis suggested that β-CD-pristane might cause cell damage through glutathione metabolism pathways and inflammatory response. After the intervention with GSH, the proliferation of MLE-12 cells was restored(P<0.05), and the intracellular ROS levels were significantly decreased(P<0.05), and the percentage of gross lung hemorrhage in pristane-induced DAH mice and DAH scores were significantly decreased(P<0.05), and the body weight was significantly increased(P<0.05), and the levels of TNF-α, IL-6 and IL-1β in BALF were significantly decreased(P<0.05). Conclusion GSH intervention can alleviate pristane-induced DAH and inflammatory response in mice. The mechanism of action may be associated with the inhibition of oxidative stress in alveolar epithelial cells and the promotion of proliferation of alveolar epithelial cells. |
| Key words: Systemic lupus erythematosus-associated diffuse alveolar hemorrhage(SLE-DAH) Reduced glutathione(GSH) Oxidative stress Inflammatory response |
