局部缺血对mTOR信号通路的影响

发布时间：2018-06-13 02:06

本文选题：小鼠 + 皮肤缺血　；参考：《重庆医科大学》2012年硕士论文

【摘要】：第一部分小鼠皮肤局部缺血模型的建立目的：研究建立稳定的昆明小鼠皮肤局部缺血模型。方法：取40只雌性昆明小鼠,随机分为A/B两组,每组20只。分别在小鼠背部建立深达皮肤筋膜层的U型皮瓣。A组为3.3×1.5cm皮瓣组,B组为2.5×1.5cm皮瓣组。皮瓣游离缘垫以医用橡胶,缝合切口,造成皮肤缺血模型,术后观察皮瓣发绀水肿及坏死情况,并用散斑全景实时血流成像仪(Moor FLPI)检测皮瓣血流。结果：肉眼观察,3.3×1.5cm皮瓣组在术后第2天,50%小鼠皮瓣末端坏死,坏死面积达皮瓣面积的(37.20±4.83)%。术后第3天100%小鼠皮瓣末端坏死。并随时间的延长,坏死面积逐渐扩大。2.5×1.5皮瓣组在术后第1天皮瓣出现发绀,水肿,发绀水肿面积占(77.46±4.51)%,术后第3、第5天发绀水肿区面积逐渐减小,至术后第7天,发绀、水肿完全消退。散斑全景实时血流成像仪(Moor FLPI)检测皮肤血流显示,术后第1天皮瓣显著性缺血(p0.05),术后第7天血流基本恢复到术前水平。结论：成功建立了昆明小鼠皮肤局部缺血模型,该模型具有操作简单、成本低、稳定、有效等优点。第二部分局部缺血对mTOR信号通路的影响目的：通过研究缺血对组织中P-4E-BPl,P-p70S6K,P-S6蛋白表达的影响,探讨缺血对蛋白质翻译mTOR信号通路的影响。方法：将30只昆明小鼠随机分为6组(每组5只)：正常对照组(即缺血0分钟组),缺血15分钟组,缺血30分钟组,缺血1小时组,缺血4小时组,缺血24小时组。除对照组外,分别在其余各实验组小鼠背部建立2.5×1.5cm U型皮瓣缺血模型。蛋白质免疫印迹法检测不同缺血时相点组织中P-4E-BP1,P-p70S6K,P-S6蛋白的表达。结果：在缺血30分钟及1小时时,组织中P-p70S6K,P-S6蛋白的表达量均明显高于正常组(P0.05)；在缺血15分钟、30分钟及1小时时,组织中P-4E-BP1蛋白的表达量表达明显高于正常组(P0.05)；在缺血24小时时,组织中P-4E-BP1、 P-p70S6K、P-S6的表达量均明显低于正常组(P0.05或P0.01)。结论：暂时性缺血(缺血时间小于1小时)可增加4E-BP1、p70S6K、S6蛋白的磷酸化,促进蛋白质翻译；而较长时间缺血(缺血24小时)可抑制4E-BP1、p70S6K、S6蛋白的磷酸化,抑制蛋白质翻译。
[Abstract]:Objective: to establish a stable skin ischemia model in Kunming mice. Methods: 40 female Kunming mice were randomly divided into two groups: group A / R B (n = 20). U type skin flaps with deep skin fascia layer were established on the back of mice respectively. Group A was 3.3 脳 1.5cm flap group and group B was 2.5 脳 1.5cm flap group. The skin ischemia model was established by using medical rubber and suture incision. The cyanotic edema and necrosis of the flap were observed after operation. The blood flow of the flap was detected by the real time speckle panoramic flow imager (moor FLPI). Results: on the second day after operation, 50% of the skin flaps in the 3. 3 脳 1.5cm skin flap group were necrotic, and the necrotic area was 37.20 卤4. 83% of the flap area. On the third day after operation, 100% of the skin flaps were necrotic. With the extension of time, the necrotic area gradually expanded. 2.5 脳 1.5 flap group showed cyanosis, edema and cyanosis on the first day after operation. The area of cyanotic edema was 77.46 卤4.51, and the area of cyanotic edema gradually decreased on the 3rd and 5th day after operation, and cyanosis occurred on the 7th day after operation. The edema completely subsided. Spot panoramic real time flow imager (moor FLPI) was used to detect the skin blood flow. On the first day after operation, the skin flap had significant ischemia (p0.05), and on the 7th day after operation, the blood flow was basically recovered to the preoperative level. Conclusion: the skin ischemia model of Kunming mice has been successfully established, which has the advantages of simple operation, low cost, stable and effective. Part two effects of local ischemia on mTOR signaling pathway. Objective: to investigate the effects of ischemia on the expression of P-4E-BPlP P-p70S6KTOR signal pathway. Methods: thirty Kunming mice were randomly divided into 6 groups (5 rats in each group): normal control group (0 minutes ischemia group, 15 minutes ischemia group, 30 minutes ischemia group, 1 hour ischemia group, 4 hours ischemia group, 24 hours ischemia group). In addition to the control group, 2. 5 脳 1.5cm U skin flap ischemia models were established in each experimental group. The expression of P-4E-BP1C P-p70S6KP6 protein was detected by Western blot. Results: the expression of P-p70S6KTP-S6 protein was significantly higher than that of normal group at 30 minutes and 1 hour after ischemia, and the expression of P-4E-BP1 protein in tissue was significantly higher than that in normal group at 15 minutes, 30 minutes and 1 hour after ischemia. After 24 hours of ischemia, the expression of P-4E-BP1 and P-p70S6KTP-S6 was significantly lower than that of normal group (P0.05 or P0.01). Conclusion: transient ischemia (ischemia time less than 1 hour) can increase the phosphorylation of 4E-BP1p70S6KS6 protein and promote protein translation, while prolonged ischemia (24 hours ischemia) can inhibit the phosphorylation of 4E-BP1p70S6KS6 protein and inhibit protein translation.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R-332;R622

【参考文献】