硫化氢抑制NLRP3调节脓毒症小鼠急性肺损伤的作用机制  

Hydrogen sulfide modulates acute lung injury in sepsis by inhibiting NLRP3 inflammasome activation in mice

作　　者：钱会涛 陈齐红 Qian Huitao;Chen Qihong(Department of Critical Care Medicine,Jiangdu People′s Hospital Affiliated to Yangzhou University,Yangzhou 225200,China)

机构地区：[1]扬州大学附属江都人民医院重症医学科,扬州225200

出　　处：《中华结核和呼吸杂志》2025年第2期130-137,共8页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家自然科学基金(81670065);江苏省卫生健康委科研项目(Z2022008);扬州市卫生健康委员会科研项目(2023-2-27)。

摘　　要：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(1μg/ml)和GYY4137(25μmol/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Objective To investigate the effects and mechanisms of the hydrogen sulfide donor GYY4137 on acute lung injury in sepsis.Methods An 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,1μg/ml)and GYY4137(25μmol/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

关 键 词：硫化氢 脓毒症 急性肺损伤 NLRP3炎症小体 

分 类 号：R459.7[医药卫生—急诊医学] R563.8[医药卫生—治疗学]