Conclusion
Defe-roxamin can promote the wound healing of rats after ischemia and hypoxia, and the effect of rapamycin is opposite. It may be related to the existence of mTOR and HIF-1 signaling pathway in chronic ischemia-hypoxia wound. 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，mTOR）、缺氧诱导因子 1α（hypoxia inducible factor 1α，HIF-1α）、VEGF mRNA 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 mTOR mRNA 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 mRNA 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 mTOR 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 mTOR 与 HIF-1 信号调节通路有关。.
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 intra-peritoneally 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 (mTOR), hypoxia inducible factor 1α (HIF-1α), and vascular endothelial growth factor (VEGF) by real-time fluorescence quantitative PCR and Western blot at 2 days after wound healing.
Objective
To explore the effect and mechanism of rapamycin and deferoxamin on wound healing after ischemia and hypoxia.
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 mTOR 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-1α 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 in groups C and D were significantly decreased when compared with the expressions in groups A and B ( P<0.05), but there was no significant difference between groups C and D ( P>0.05). The relative expressions of HIF-1α protein in groups A, B, and C were significantly increased when compared with expression in group D ( P<0.05), but there was no significant difference between groups A, B, and C ( P>0.05). The relative expression of VEGF protein were significantly lower in groups B, C, and D than group A, in group D than groups B and C, and in group C than group B ( P<0.05).