HSF1在小鼠呼吸机相关性肺损伤内源性保护机制中的作用:与HMGB1的关系  

Role of HSF1 in endogenous protective mechanism underlying mechanical ventilator-induced lung injury in mice:relationship with HMGB1

作　　者：徐兴桂 牟传琳[1] 孙莉莉 毕夏 孙立新[1] 王明山[1] 马福国[1] 韩伟[3] Xu Xinggui;Mu Chuanlin;Sun Lili;Bi Xia;Sun Lixin;Wang Mingshan;Ma Fuguo;Han Wei(Department of Anesthesiology,Qingdao Municipal Hospital,Qingdao 266071,China;Department of Nephrology and Immunology,Qingdao Women and Children′s Hospital,Qingdao 266034,China;Department of Respiratory and Critical Care Medicine,Qingdao Municipal Hospital,Qingdao 266071,China)

机构地区：[1]青岛市市立医院麻醉科,青岛266071 [2]青岛市妇女儿童医院肾脏免疫科,青岛266034 [3]青岛市市立医院呼吸与危重症医学科,青岛266071

出　　处：《中华麻醉学杂志》2023年第2期210-215,共6页Chinese Journal of Anesthesiology

基　　金：山东省医学会科研基金(YXH2022ZX02107)。

摘　　要：目的评价热休克转录因子1(HSF1)在小鼠呼吸机相关性肺损伤(VILI)内源性保护机制中的作用及其与高迁移率族蛋白B1(HMGB1)的关系。方法SPF级健康C57BL/6雄性小鼠40只,6~8周龄,体质量20~25 g,采用随机数字表法分为4组(n=10):对照组(C组)、VILI组、阴性对照siRNA+VILI组(NV组)和HSF1 siRNA+VILI组(siRNA组)。机械通气前48 h时,NV组气道内注射阴性对照siRNA 5 nmol,siRNA组气道内注射HSF1 siRNA 5 nmol,均用无菌磷酸盐缓冲液稀释至50μl。VILI组、NV组和siRNA组机械通气(V_(T) 35 ml/kg,RR 75次/min,I∶E 1∶2,FiO_(2)21%)4 h构建小鼠VILI模型,C组仅行气管切开,保留自主呼吸。分别于气管插管后即刻和机械通气4 h时,行动脉血气分析,记录PaO_(2);然后深麻醉下处死小鼠,收集支气管肺泡灌洗液(BALF)和肺组织,采用ELISA法测定BALF中TNF-α、IL-1β、HMGB1浓度,测定肺组织湿重/干重(W/D)比值,HE染色法观察肺组织病理学结果,并行肺损伤评分,采用qRT-PCR法检测肺组织HMGB1和HSF1的mRNA表达,Western blot法检测肺组织HMGB1和HSF1的表达。结果与C组比较,VILI组、NV组和siRNA组机械通气4 h时PaO_(2)降低,BALF中TNF-α、IL-1β、HMGB1浓度、肺组织W/D比值、肺损伤评分、HMGB1及其mRNA的表达水平升高(P<0.05或0.01);与VILI组和NV组比较,siRNA组机械通气4 h时PaO_(2)降低,BALF中TNF-α、IL-1β、HMGB1浓度、肺组织W/D比值、肺损伤评分、HMGB1及其mRNA表达水平升高,HSF1及其mRNA表达下调(P<0.05或0.01)。VILI组和NV组上述各指标比较差异无统计学意义(P>0.05)。结论HSF1参与了小鼠VILI内源性保护机制,可能与下调HMGB1的表达,减轻肺组织炎症反应有关。Objective To evaluate the role of heat shock transcription factor 1(HSF1)in the endogenous protective mechanism underlying mechanical ventilator-induced lung injury(VILI)in mice and the relationship with high mobility group box 1(HMGB1).Methods Forty SPF healthy male C57BL/6 mice,aged 6-8 weeks,weighing 20-25 g,were divided into 4 groups(n=10 each)by the random number table method:control group(group C),VILI group(group VILI),negative control siRNA+VILI group(group NV)and HSF1 siRNA+VILI group(group siRNA).At 48 h before mechanical ventilation,negative control siRNA 5 nmol and HSF1 siRNA 5 nmol were intratracheally injected in NV and siRNA groups respectively,and the solution was diluted to 50μl with the sterile phosphate buffer in both groups.Group C kept spontaneous breathing for 4 h,and the rest animals were mechanically ventilated(tidal volume 35 ml/kg,respiratory rate 75 breaths/min,inspiratory/expiratory ratio 1∶2,fraction of inspired oxygen 21%)for 4 h.Blood samples from the femoral artery were collected for arterial blood gas analysis immediately after endotracheal intubation and at 4 h of ventilation,and PaO_(2) was recorded.Then the mice were sacrificed under deep anesthesia to collect lung tissues and bronchoalveolar lavage fluid(BALF).The concentrations of interleukin-1beta(IL-1β),tumor necrosis factor-alpha(TNF-α)and HMGB1 in BALF were determined by enzyme-linked immunosorbent assay.The pathological results were observed by hematoxylin-eosin staining,and lung injury was assessed and scored.The wet/dry(W/D)weight ratio of lung tissues was calculated.The expression of HMGB1 and HSF1 mRNA in lung tissues(by quantitative real-time polymerase chain reaction)and expression of HMGB1 and HSF1 protein in lung tissues(by Western blot)were determined.Results Compared with group C,PaO_(2) was significantly decreased at 4 h of ventilation,the concentrations of TNF-α,IL-1βand HMGB1 in BALF,W/D ratio and lung injury score were increased,and the expression of HMGB1 protein and mRNA in lung tissues was up-regu

关 键 词：热休克转录因子 呼吸机相关性肺损伤 高迁移率族蛋白质类 

分 类 号：R563[医药卫生—呼吸系统]