雷帕霉素及去铁敏对缺血缺氧创面愈合的影响  

Effects of rapamycin and deferoxamin on wound healing after ischemia and hypoxia

作　　者：聂开瑜[1] 胡鹏[1] 王达利[1] 魏在荣[1] 曾学琴 孙广峰[1] 

机构地区：[1]遵义医学院附属医院烧伤整形外科,贵州遵义563000

出　　处：《中国修复重建外科杂志》2017年第6期718-722,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：贵州省科学技术基金(黔科合J字[2011]2266号)~~

摘　　要：目的探讨雷帕霉素及去铁敏对缺血缺氧创面愈合的影响及其作用机制。方法 SPF级雄性成年SD大鼠40只,体质量(300±20)g,于背部制备缺血缺氧创面模型;将其随机分为4组(n=10),分别为空白对照组(A组)、去铁敏干预组(B组)、雷帕霉素干预组(C组)、去铁敏+雷帕霉素共同干预组(D组)。模型制备后3、6、9 d,A、B、C、D组分别腹腔注射生理盐水、去铁敏(10 mg/kg)、雷帕霉素(3 mg/kg)、去铁敏(10 mg/kg)+雷帕霉素(3 mg/kg)。模型制备后观察创面愈合情况并记录愈合时间;于创面完全愈合后第2天切取创面愈合组织,采用实时荧光定量PCR及Western blot检测创面组织中雷帕霉素靶蛋白(mammalian target of rapamycin,m TOR)、缺氧诱导因子1α(hypoxia inducible factor 1α,HIF-1α)、VEGF m RNA及蛋白的表达。结果各组大鼠均成活至实验完成,创面均愈合;其中A、B、D组创面愈合时间均较C组明显缩短(P<0.05);A、B、D组间比较差异无统计学意义(P>0.05)。实时荧光定量PCR检测示,C、D组m TOR m RNA表达较A、B组明显下调(P<0.05),A、B组间比较差异有统计学意义(P<0.05),C、D组间比较差异无统计学意义(P>0.05)。B、D组HIF-1α、VEGF m RNA表达较A、C组明显上调,A组较C组表达上调,比较差异有统计学意义(P<0.05);B、D组间比较差异无统计学意义(P>0.05)。Western blot检测示,C、D组m TOR蛋白相对表达量较A、B组明显下调(P<0.05),C、D组间比较差异无统计学意义(P>0.05)。A、B、C组HIF-1α蛋白相对表达量明显高于D组(P<0.05),A、B、C组间比较差异无统计学意义(P>0.05)。B、C、D组VEGF蛋白相对表达量较A组明显下调,D组低于B、C组,C组低于B组,比较差异均有统计学意义(P<0.05)。结论去铁敏可促进大鼠缺血缺氧创面愈合,而雷帕霉素作用相反;可能与慢性缺血缺氧创面中存在m TOR与HIF-1信号调节通路有关。Objective To explore the effect and mechanism of rapamycin and deferoxamin on wound healing after ischemia and hypoxia. Methods The model of ischemia and hypoxia wound was made on the back of 40 SPF male adult Sprague Dawley rats, weight （300±20） g; they were randomly divided into 4 groups （n=10）： the control group （group A）, deferoxamine intervention group （group B）, rapamycin intervention group （group C）, and deferoxamine＋rapamycin intervention group （group D）. At 3, 6, and 9 days after model preparation, rats of groups A, B, C, and D were intraperitoneally injected with normal saline, deferoxamin （10 mg/kg）, rapamycin （3 mg/kg）, deferoxamin （10 mg/kg）＋ rapamycin （3 mg/kg） respectively. The wound healing was observed and the healing time was recorded in each group; the wound healing tissue was harvested to test the mRNA and protein expressions of mammalian target of rapamycin （roTOR）, hypoxia inducible factor let （HIF-1α）, and vascular endothelial growth factor （VEGF） by real-time fluorescence quantitative PCR and Western blot at 2 days after wound healing. Results All rats survived to the end of the experiment, and wounds healed; the healing time of groups A, B, and D was significantly shorter than that of group C （P〈0.05）, but there was no significant difference between groups A, B, and D （P〉0.05）. Real-time fluorescence quantitative PCR showed that the expression of roTOR mRNA in groups C and D was significantly decreased when compared with the expressions in groups A and B （P〈0.05）; there was significant difference between groups A and B （P〈0.05）, but no significant difference between groups C and D （P〉0.05）. The expressions of HIF-lct mRNA and VEGF mRNA were signi- ficantly higher in groups B and D than groups A and C, and in group A than group C （P〈0.05）, but there was no signifi- cant difference between groups B and D （P〉0.05）. Western blot showed that the relative expressions of mTOR protein i

关 键 词：雷帕霉素 去铁敏 雷帕霉素靶蛋白 缺氧诱导因子1Α 创面愈合 

分 类 号：R622[医药卫生—整形外科]