高氧诱发幼鼠急性肺损伤时Wnt/β-catenin信号通路与自噬的关系  

Relationship between Wnt/β-catenin signaling pathway and autophagy during hyperoxia-induced acute lung injury in infantile rats

作　　者：贾金娥 王建波[1] 于志强[1] Jia Jine;Wang Jianbo;Yu Zhiqiang(Department of Anesthesiology,Tianjin Central Hospital of Obstetrics and Gynecology,Tianjin 300052,China)

机构地区：[1]天津市中心妇产科医院麻醉科,天津300052

出　　处：《中华麻醉学杂志》2023年第10期1232-1236,共5页Chinese Journal of Anesthesiology

基　　金：天津市卫生健康科技项目 (TJWJ2022MS042)。

摘　　要：目的评价高氧诱发幼鼠急性肺损伤时Wnt/β-连环蛋白(β-catenin)信号通路及与自噬的关系。方法清洁级健康雄性SD幼鼠24只,14日龄,体质量40~50 g,采用随机数字表法分为4组(n=6):对照组(C组)、高氧诱发急性肺损伤(HALI)组、HALI+IWP-2组(HI组)和HALI+DMF组(HD组)。采用吸入高浓度氧气(氧浓度>90%)72 h的方法制备HALI模型。于制模前30 min,HI组腹腔注射IWP-215 mg/kg,HD组腹腔注射相同容量DMF溶剂,C组与HALI组腹腔注射相同容量生理盐水。各组于每日同一时间点给药,连续给药3 d。制模结束后,经颈总动脉取血标本行血气分析,计算氧合指数(OI);然后深麻醉下处死幼鼠,取肺组织,测定湿重/干重(W/D)比值,光镜下观察肺组织病理学结果并行肺损伤评分,采用ELISA法测定IL-6、IL-1β和TNF-α的含量,Western blot法检测Wnt3a、β-catenin、微管相关蛋白轻链3(LC3)、Beclin1和p62的表达,计算LC3Ⅱ/LC3Ⅰ比值。结果与C组比较,HALI组、HI组和HD组OI降低,肺组织W/D比值、肺损伤评分、IL-6、IL-1β和TNF-α含量升高,Wnt3a、β-catenin和Beclin1表达上调,p62表达下调,LC3Ⅱ/LC3Ⅰ比值升高(P<0.05);与HALI组比较,HI组OI降低,肺组织W/D比值、肺损伤评分、IL-6、IL-1β和TNF-α含量升高,Wnt3a、β-catenin和p62表达下调,Beclin1表达上调,LC3Ⅱ/LC3Ⅰ比值升高(P<0.05),HD组上述指标差异无统计学意义(P>0.05);与HI组比较,HD组OI升高,肺组织W/D比值、肺损伤评分、IL-6、IL-1β和TNF-α含量降低,Wnt3a、β-catenin和p62表达上调,Beclin1表达下调,LC3Ⅱ/LC3Ⅰ比值降低(P<0.05)。结论Wnt/β-catenin信号通路可能通过对自噬的负向调控参与幼鼠HALI的过程。Objective To evaluate the relationship between Wnt/β-catenin signaling pathway and autophagy during hyperoxia-induced acute lung injury in infantile rats.Methods A total of 24 clean-stage healthy male infantile Sprague-Dawley rats,aged 14 days,weighing 40-50 g,were allocated into 4 groups(n=6 each)using a random number table method:control group(C group),hyperoxic acute lung injury(HALI)group,HALI+IWP-2 group(HI group)and HALI+DMF group(HD group).HALI model was developed by inhaling oxygen at a concentration greater than 90%for 72 h.Starting from 30 min before developing the model,IWP-215 mg/kg was intraperitoneally injected every day for 3 consecutive days in HI group,the equal volume of DMF solution was injected every day for 3 consecutive days in HD group,and the equal volume of normal saline was intraperitoneally injected instead in C and HALI groups.Blood samples were taken from the common carotid artery for blood gas analysis at the end of developing the model,and oxygenation index(OI)was calculated.Then the infantile rats were sacrificed under deep anesthesia,and lungs were removed for examination of the pathological changes which were scored and for determination of the weight to dry weight ratio(W/D ratio),contents of interleukin-6(IL-6),IL-1β,and tumor necrosis factor-alpha(TNF-α)(by enzyme-linked immunosorbent assay)and expression of Wnt3a,β-catenin,microtubule-associated protein 1 light chain 3(LC3),Beclin1 and p62(by Western blot).LC3Ⅱ/LC3Ⅰratio was calculated.Results Compared with C group,the OI was significantly decreased,the W/D ratio,lung injury score and contents of IL-6,IL-1βand TNF-αwere increased,and the expression of Wnt3a,β-catenin and Beclin1 was up-regulated,the expression of p62 was down-regulated,and LC3Ⅱ/LC3Ⅰratio was increased in HALI,HD and HI groups(P<0.05).Compared with HALI group,the OI was significantly decreased,the W/D ratio,lung injury score and contents of IL-6,IL-1βand TNF-αwere increased,and the expression of Wnt3a,β-catenin and p62 was down-regulated,the

关 键 词：氧 急性肺损伤 WNT蛋白质类 Β连环蛋白 自噬 

分 类 号：R614[医药卫生—麻醉学]