信号转导与转录激活因子6通过调控铁死亡介导脓毒症小鼠骨骼肌细胞损伤  

Signal transducer and activator of transcription 6 mediates skeletal muscle cell injury in septic mice by regulating ferroptosis

作　　者：李福星[1] 王舒[1] 周仪华[1] 龚园其[1] Li Fuxing;Wang Shu;Zhou Yihua;Gong Yuanqi(Department of Intensive Care Unit,the Second Affiliated Hospital of Nanchang University,Nanchang 330006,Jiangxi,China)

机构地区：[1]南昌大学第二附属医院重症医学科,江西南昌330006

出　　处：《中华危重病急救医学》2023年第8期813-817,共5页Chinese Critical Care Medicine

基　　金：江西省自然科学基金(20202BAB206058)。

摘　　要：目的:探讨在脓毒症模型中信号转导与转录激活因子6(STAT6)对骨骼肌细胞铁死亡的影响及潜在机制。方法:将24只8周龄雄性SPF级昆明小鼠按随机数字表法分为正常对照组、假手术组、脓毒症模型组、STAT6抑制剂预处理组,每组6只。采用盲肠结扎穿孔术(CLP)构建小鼠脓毒症模型;假手术组开腹暴露盲肠后关腹。STAT6抑制剂预处理组于制模前1 h腹腔注射AS1517499溶液10 mg/kg;假手术组及模型组腹腔注射等量生理盐水。正常对照组小鼠不予任何手术及药物干预。于制模后24 h处死小鼠取后肢肌肉组织,苏木素-伊红(HE)染色后,光镜下观察组织病理学改变;醋酸铀-枸橼酸铅双染后,透射电镜下观察线粒体形态变化;采用蛋白质免疫印迹试验(Western blotting)检测肌肉组织中铁死亡标志物蛋白几丁质酶-3样蛋白1(CHI3L1)、环氧合酶-2(COX-2)、酰基辅酶A合成酶长链家族成员4(ACSL4)、铁蛋白重链1(FTH1)、谷胱甘肽过氧化物酶4(GPx4)的表达水平。结果:光镜下显示,正常对照组和假手术组小鼠骨骼肌形态结构正常;模型组骨骼肌结构疏松,肌纤维变小及萎缩,炎症细胞浸润,甚至出现肌纤维缺失;与模型组比较,STAT6抑制剂预处理组骨骼肌结构紧密,骨骼肌萎缩情况有所改善。透射电镜下显示,正常对照组和假手术组骨骼肌细胞超微形态正常;模型组骨骼肌超微形态学特征显示,细胞膜断裂和出泡,线粒体变小、膜密度增高、线粒体嵴减少或消失、线粒体外膜断裂,细胞核大小正常但缺乏染色质凝聚;与模型组比较,STAT6抑制剂预处理组肌细胞超微结构损伤有所改善。与正常对照组和假手术组比较,模型组小鼠肌肉组织中CHI3L1、COX-2、ACSL4、FTH1的蛋白表达均明显升高,GPx4蛋白表达明显下降,说明脓毒症小鼠模型骨骼肌细胞出现特征性线粒体损伤,同时铁死亡标志物蛋白表达异常;与模型组相比,STAT6抑制剂预�Objective To explore the effect of signal transducer and activator of transcription 6(STAT6)on ferroptosis in skeletal muscle cells in sepsis model and its potential mechanism.Methods Twenty-four 8-week-old male specific pathogen free Kunming mice were divided into normal control group,sham group,sepsis model group and STAT6 inhibitor pretreatment group according to random number table method with 6 mice in each group.A mouse sepsis model was reproduced by cecal ligation and perforation(CLP).In the sham group,the skin of mice was sutured after exposing the cecum tissue.In the STAT6 inhibitor pretreatment group,10 mg/kg AS1517499 was injected intraperitoneally 1 hour before model reproduction.The sham group and the model group were intraperitoneally injected with the same volume of normal saline.Mice in the normal control group did not receive any operation or drug intervention.The mice were sacrificed 24 hours after model reproduction,and the muscle tissue of hind limb was obtained under sterile condition.Hematoxylin-eosin(HE)staining was used to observe the histopathology with optical microscope,and mitochondrial morphological changes were observed by transmission electron microscopy after double staining with uranium acetate lead citrate.The ferroptosis marker proteins expressions of chitinase-3-like protein 1(CHI3L1),cyclooxygenase-2(COX-2),acyl-CoA synthetase long-chain family member 4(ACSL4),ferritin heavy chain 1(FTH1),and glutathione peroxidase 4(GPx4)were detected by Western blotting.Results Under the optical microscope,the morphology and structure of skeletal muscle tissues in the normal control and sham groups were normal.In the model group,the structure of skeletal muscle tissues was loose,the muscle fiber became smaller and atrophic,inflammatory cell infiltration and even muscle fiber loss were found.Compared with the model group,the structure of skeletal muscle tissues was tight and skeletal muscle atrophy was improved in the STAT6 inhibitor pretreatment group.The ultrastructure of skeletal muscle ce

关 键 词：脓毒症所致肌病 铁死亡 信号转导与转录激活因子6 几丁质酶-3样蛋白1 

分 类 号：R459.7[医药卫生—急诊医学]