自噬在深二度烧伤创面早期加深中的表达及作用

发布时间：2018-06-04 21:49

本文选题：烧伤 + 自噬　；参考：《中国人民解放军医学院》2013年硕士论文

【摘要】：目的：探讨自噬在深二度烧伤创面早期加深中的表达及作用。方法：采用220-250g雄性Wistr大鼠作为研究对象，实验分两部分，第一部分观察大鼠深二度烧伤后创面自噬的表达规律，探讨自噬与深二度创面加深的可能联系：Wistar大鼠背部制作深二度烫伤模型，烫伤面积10%，观察烫伤后1h、6h、12h、24h、48h、72h创面组织自噬标志蛋白LC3、Beclin-1的表达及变化规律；HE染色及Masson染色显示创面组织形态和创面深度的变化。第二部分研究雷帕霉素对大鼠深二度烧伤创面自噬及创面早期加深的影响：Wistar大鼠背部制作深二度烫伤模型，烫伤面积10%，治疗组腹腔注射雷帕霉素，对照组腹腔注射等量溶媒，观察两组烫伤后6h、1d、2d、3d创面组织自噬体，自噬标志蛋白LC3、Beclin-1的表达及变化规律；TUNEL法检测创面凋亡；同时检测烧伤后创面局部多普勒血流（LDF）值，，创面组织中IL-8、Na-K-ATP酶、髓过氧化物酶(MPO)活性和丙二醛(MDA)的含量变化，HE染色及Masson染色显示创面组织坏死及加深程度。健康大鼠组不做任何处理，作为健康对照。结果：第一部分实验结果：烫伤后创面组织自噬水平持续下降，至伤后24h达到最低，此后稍有升高，但仍远低于正常水平；创面组织真皮深层自噬被激活；烫伤后创面深度随着时间的推移进行性加深。第二部分实验结果：治疗组与对照组相比，自噬体明显增多，LC3、Beclin-1表达明显增高，创面凋亡率明显低于对照组；治疗组创面LDF值及Na-K-ATP酶活性比对照组明显升高，IL-8、MPO活性和MDA的含量较对照组明显下降，创面深度较对照组减轻。结论：1.大鼠深二度烫伤后早期创面组织自噬减少，凋亡增加，可能是参与创面加深的因素；真皮深层自噬的激活，可能是一种保护机制。2.雷帕霉素使大鼠深二度烧伤创面自噬增强，抑制烧伤创面早期加深。
[Abstract]:Objective: to investigate the expression and role of autophagy in the early deepening of deep second degree burn wounds. Methods: 220-250g male Wistr rats were used as the research objects. The experiment was divided into two parts. The first part was to observe the expression of autophagy after deep second degree burn in rats. To explore the possible connection between autophagy and deep second degree wound deepening, the model of deep second degree scald was made on the back of Wistar rats. The expression and changes of autophagy marker protein LC3 and Beclin-1 in wound tissue were observed at 1 h, 6 h, 12 h, 24 h and 72 h after scalding. The changes of tissue morphology and depth of wound were observed by HE staining and Masson staining. The second part was to study the effect of rapamycin on the autophagy of deep second degree burn wound and the early deepening of wound in rats. The rat model of deep second degree scalding was made in the back of rats. The scalded area was 10. The treatment group was injected with rapamycin intraperitoneally, and the control group was injected with the same amount of solvent. The expression of autophagy, the expression of autophagy protein LC3 and Beclin-1 and the Tunel method were used to detect the wound apoptosis, and the local Doppler blood flow (LDF) and IL-8 Na-K-ATPase in the wound tissue were measured. The changes of myeloperoxidase (MPO) activity and malondialdehyde (MDA) content in the wound showed the degree of necrosis and deepening of wound tissue by HE staining and Masson staining. The healthy rats group did not do any treatment, as a healthy control. Results: the results of the first part of the experiment were as follows: the level of autophagy of wound tissue decreased continuously after scald, reached the lowest at 24 hours after burn, then increased slightly, but was still far below the normal level, the deep dermal autophagy of wound tissue was activated. The depth of burn wound deepened with time. The second part of the experimental results: compared with the control group, autophagy significantly increased the expression of LC3 / Beclin-1, and the rate of wound apoptosis was significantly lower than that of the control group. Compared with the control group, the LDF value and the activity of Na-K-ATP enzyme in the treatment group were significantly higher than those in the control group. The activity of IL-8 MPO and the content of MDA in the treatment group were significantly lower than those in the control group. Conclusion 1. The decrease of autophagy and the increase of apoptosis in the early stage after deep second degree scalding may be the factors involved in the deepening of the wound, and the activation of deep dermal autophagy may be a protective mechanism. Rapamycin enhanced the autophagy of deep second degree burn wound and inhibited the early deepening of burn wound.
【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R644

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