目的探究硫化氢缓释剂GYY4137在脓毒症急性肺损伤中的作用及调节机制。
方法使用盲肠结扎术（CLP）构建脓毒症小鼠急性肺损伤模型。手术结束30 min后，按50 mg/kg剂量腹腔注射GYY4137或者等体积0.9%氯化钠溶液。将C57BL/6J小鼠按随机数字表法随机分为4组，即假手术组（Sham组）、假手术+GYY4137组（GYY4137组）、脓毒症组（CLP组）、脓毒症+GYY4137组（CLP+GYY4137组）。小动物全身体积描记系统检测小鼠呼吸参数分钟通气量和呼气中期流速。HE观察肺组织病理损伤、免疫荧光检测炎症细胞、实时定量PCR检测白细胞介素（IL）-1β、IL-6、血管内皮钙黏蛋白、细胞间黏附分子1、血管细胞黏附分子1的mRNA表达、Western blotting检测NOD样受体家族pyrin结构域3（NLRP3）、Pro-IL-1β、IL-1β和环磷酸鸟苷-腺苷合成酶（cGAS）/干扰素基因刺激因子（Sting）/核因子-κB（NF-κB）信号轴蛋白表达。体外培养小鼠肺微血管内皮细胞，利用脂多糖（1 μg/ml）和GYY4137（25 μmol/L）构建脓毒症损伤及探究硫化氢作用机制的细胞模型。实时定量PCR检测细胞炎症因子及黏附分子表达。使用Shapiro-Wilk test验证数据是否符合正态分布，并通过 Brown-Forsythe test 验证方差齐性。 P<0.05 表示差异有统计学意义。
结果（1）与Sham组相比，CLP组小鼠呼吸参数分钟通气量［（36.32±3.91）ml/min］和呼气中期流速［（1.43±0.26）ml/s］分别均低于Sham组［（50.14±6.07）ml/min和（2.70±0.46）ml/s］，CLP+GYY4137组分钟通气量［（45.83±2.33）ml/min］和呼气中期流速［（2.02±0.16）ml/s］均高于CLP组，差异均有统计学意义（均 P<0.05）。与Sham组相比，CLP组小鼠肺组织损伤明显，肺泡塌陷、肺组织间隔增厚、渗出更多，肺组织损伤评分平均为4分，高于Sham组的1分，CLP+GYY4137组肺损伤明显减轻，评分为2.3分，低于CLP组，差异均有统计学意义（均 P<0.05）。（2）CLP组炎症细胞浸润、炎症因子及黏附分子mRNA水平均高于Sham组，CLP+GYY4137组均低于CLP组，差异均有统计学意义（均 P<0.05）。（3）CLP组NLRP3、Pro-IL-1β、IL-1β、cGAS、Sting、NF-κB蛋白表达均高于Sham组（均 P<0.05），CLP+GYY4137组均低于CLP组，差异均有统计学意义（均 P<0.05）。（4）脂多糖刺激的肺内皮炎症因子和黏附分子水平均高于对照组，GYY4137和脂多糖共处理组均低于脂多糖组，差异均有统计学意义（均 P<0.05）。
结论硫化氢缓释剂GYY4137通过cGAS/Sting/NF-κB信号通路抑制NLRP3炎症小体激活，调节脓毒症小鼠急性肺损伤的炎症反应。
ObjectiveTo investigate the effects and mechanisms of the hydrogen sulfide donor GYY4137 on acute lung injury in sepsis.
MethodsAn acute lung injury model was established using the method of cecal ligation and puncture(CLP). Mice received an intraperitoneal injection of GYY4137 (50 mg/kg) or saline 30 minutes after surgery. C57BL/6J mice were divided into four groups: the sham operation group (Sham), the sham operation with GYY4137 group (GYY4137), the sepsis group (CLP), and the sepsis with GYY4137 group (CLP+GYY4137). Respiratory parameters (minute ventilation volume and expiratory flow 50) were measured using a whole-body plethysmography system. Lung tissue was evaluated by hematoxylin and eosin (H&E) staining, and inflammatory cells were identified by immunofluorescence. mRNA expression of inflammatory factors (interleukin-1β, interleukin-6) and adhesion molecules (vascular endothelial cadherin, intercellular cell adhesion molecule-1, vascular cell adhesion molecule-1) were quantified by real-time quantitative polymerase chain reaction (RT-PCR). Levels of NLRP3, Pro-IL-1β, IL-1β, and the cyclic guanosine monophosphate synthase (cGAS)/stimulator of interferon genes (Sting)/NF-κB signaling proteins were analyzed by Western blotting. In vitro, murine pulmonary microvascular endothelial cells were cultured and exposed to lipopolysaccharide (LPS, 1 μg/ml) and GYY4137 (25 μmol/L) to simulate sepsis-induced damage and to study the mechanism of hydrogen sulfide action. Levels of inflammatory factors and adhesion molecules in cells were quantified by RT-PCR. Data were analyzed for normal distribution using Shapiro-Wilk test, then variance homogeneity by Brown-Forsythe test. P-value<0.05 was set as statistical significance for all analyses.
Results(1) MV and EF50 values were significantly lower in the CLP group [(36.32±3.91)ml/min and (1.43±0.26)ml/s, respectively] compared to the Sham group [(50.14±6.07)ml/min and (2.70±0.46)ml/s, respectively]. These parameters were higher in the CLP+GYY4137 group [(45.83±2.33)ml/min and (2.02±0.16)ml/s, respectively] compared to the CLP group ( P<0.05). (2) Severe lung tissue damage, alveolar collapse, increased septal thickening and exudation were observed in the CLP group and significantly reduced in the CLP+GYY4137 group ( P<0.05). (3) Infiltration of inflammatory cells and mRNA levels of inflammatory factors and adhesion molecules were increased in the CLP group compared to the Sham group ( P<0.05), and decreased in the CLP+GYY4137 group compared to the CLP group ( P<0.05). (4) Protein levels of NLRP3, Pro-IL-1β, IL-1β, cGAS, Sting, and NF-κB were higher in the CLP group compared to the Sham group ( P<0.05) and were reduced in the CLP+GYY4137 group compared to the CLP group ( P<0.05). (5) LPS induced higher levels of inflammatory factors and adhesion molecules in pulmonary endothelial cells compared to the control, which were reduced in cells co-treated with GYY4137 and LPS ( P<0.05).
ConclusionThe hydrogen sulfide donor GYY4137 effectively modulates acute lung injury in septic mice by inhibiting NLRP3 inflammasome activation via the cGAS/Sting/NF-κB signaling pathway.
钱会涛,陈齐红. 硫化氢抑制NLRP3调节脓毒症小鼠急性肺损伤的作用机制[J]. 中华结核和呼吸杂志,2025,48(02)：130-137.
DOI:10.3760/cma.j.cn112147-20240718-00413版权归中华医学会所有。
未经授权，不得转载、摘编本刊文章，不得使用本刊的版式设计。
除非特别声明，本刊刊出的所有文章不代表中华医学会和本刊编委会的观点。
钱会涛：实验设计、研究实施、数据整理、统计学分析、文章撰写；陈齐红：实验设计、研究指导、论文审阅、资金支持
文档即答
智问智答
机器翻译
你好，我可以帮助您更好的了解本文，请向我提问您关注的问题。
