NLRP3炎性体在深二度烧伤创面早期加深中的作用及自噬对其的调控

发布时间：2018-03-30 15:27

本文选题：烧伤　切入点：创面加深　出处：《中国人民解放军医学院》2016年博士论文

【摘要】：目的：一、烧伤创面过度炎症反应是导致创面早期加深的重要因素之一,而NLRP3炎性体在机体固有免疫和炎症应答中发挥重要作用,但NLRP3炎性体在烧伤创面早期加深中的作用尚不清楚,本研究的第一个目的在于探讨在深Ⅱ度烧伤创面加深过程中NLRP3炎性体的表达和作用。二、自噬是一种高度保守的溶酶体依赖的降解途径,我们前期研究发现自噬对烧伤创面加深有保护作用,但具体机制尚不明了。自噬与NLRP3炎性体在深二度烧伤创面早期加深过程中存在什么样的联系,自噬是否能通过调控NLRP3炎性体介导的炎性通路来抑制烧伤创面过度炎症反应,从而发挥对烧伤创面加深的保护作用,是第二部分研究的主要内容。方法：实验分为两部分。第一部分研究NLRP3炎性体在深二度烧伤创面早期加深中的表达和作用.采用Wistar大鼠,建立深二度烫伤模型,观察烫伤后1h,24h,48h,和72h创面组织NLRP3炎性体的表达及变化规律,并使用NLRP3炎性体特异性抑制剂MNS抑制其活性,观察对创面NLRP3炎性体活性表达的影响,以及对创面炎性浸润、血流量、创面深度,以及创面愈合时间的影响。第二部分探讨自噬对烧伤创面NLRP3炎性体的调控及对创面早期加深的影响。大鼠烫伤后随机分为三组：分别给予溶媒对照、自噬增强剂雷帕霉素和自噬抑制剂氯喹进行治疗,检测各组创面组织自噬水平、NLRP3炎性体活性、炎性因子、自由基损伤和病理HE染色及Masson染色。结果：第一部分：烫伤后烧伤创面组织NLRP3炎性体活性明显增强,NLRP3炎性体主要表达于创面间生态组织的巨噬细胞。NLRP3炎性体抑制剂MNS能显著抑制创面NLRP3炎性体活性和炎性因子的表达,减少炎性浸润,增加创面血流量,减轻创面早期加深,加速创面愈合。第二部分：与对照组相比,雷帕霉素治疗组创面自噬水平明显升高,NLRP3炎性体活性显著降低,创面炎性浸润减轻,自由基损伤减少,创面加深程度减轻。而氯喹治疗组相比于对照组,其创面自噬水平降低,NLRP3炎性体活性增高,创面炎性浸润加剧,自由基损伤加重,创面加深程度更为明显。结论：第一部分：大鼠深二度烫伤后创面组织NLRP3炎性体活性增加且主要表达于间生态组织的巨噬细胞,抑制其活性可改善创面早期加深,并加速创面愈合。第二部分：激活或抑制自噬可调控创面NLRP3炎性体活性,对创面深度变化产生影响,表明自噬对烧伤创面早期加深的保护作用可通过调控创面NLRP3炎性体活性而发挥作用。
[Abstract]:Objective: first, the excessive inflammatory reaction of burn wound is one of the important factors leading to the early deepening of the wound, and the NLRP3 inflammatory body plays an important role in the innate immunity and inflammatory response of the body. However, the role of NLRP3 inflammatory body in the early stage of burn wound deepening is not clear. The first purpose of this study is to explore the expression and role of NLRP3 inflammatory body in the process of deep second degree burn wound deepening. Autophagy is a highly conserved lysosomal dependent degradation pathway. Our previous studies have shown that autophagy has a protective effect on burn wound deepening. However, the specific mechanism is not clear. What is the relationship between autophagy and NLRP3 inflammatory body in the early deepening process of deep second degree burn wound, and whether autophagy can inhibit the excessive inflammatory reaction of burn wound by regulating the inflammatory pathway mediated by NLRP3 inflammatory body. The main contents of the second part of the study were as follows: methods: the experiment was divided into two parts. The first part was to study the expression and role of NLRP3 inflammatory body in the early deepening of deep second degree burn wound. Wistar rats were used. A deep second degree scalding model was established to observe the expression and changes of NLRP3 inflammatory body in wound tissue from 24 h to 48 h and 72 h after scald. MNS, a specific inhibitor of NLRP3, was used to inhibit its activity, and the effect on the expression of NLRP3 inflammatory body activity was observed. And the inflammatory infiltration, blood flow, depth of the wound, The second part was to investigate the effect of autophagy on the NLRP3 inflammatory body of burn wound and its influence on the early deepening of burn wound. Rats were randomly divided into three groups after scalding: the control group was given solvent respectively. Autophagy enhancer rapamycin and autophagy inhibitor chloroquine were used to detect the level of autophagy in wound tissue and the activity of NLRP3 inflammatory body and inflammatory factor. Free radical injury and pathological HE staining and Masson staining. Results: part one: the activity of NLRP3 inflammatory body in burn wound tissue after scalding was significantly enhanced, mainly expressed in macrophages. NLRP3 inflammatory body inhibited in the ecological tissue of the wound. MNS could significantly inhibit the activity of NLRP3 inflammatory body and the expression of inflammatory factors. In the second part, compared with the control group, the level of wound autophagy in the rapamycin treated group was significantly increased, and the activity of NLRP3 inflammatory body was significantly decreased in the rapamycin treated group. Compared with the control group, the inflammatory infiltration, free radical injury and depth of wound in chloroquine group were decreased, the activity of NLRP3 inflammatory body was increased, the inflammatory infiltration of wound was aggravated, and the free radical injury was aggravated in chloroquine group. Conclusion: in the first part, the activity of NLRP3 inflammatory body in the wound tissue is increased and mainly expressed in the macrophages of mesenchymal tissue after deep second degree scalding. Inhibition of the activity can improve the early deepening of the wound. The second part: activation or inhibition of autophagy can regulate the activity of NLRP3 inflammatory body and influence the depth of wound. These results suggest that autophagy may play an important role in the protection of burn wound by regulating the activity of NLRP3 inflammatory body.
【学位授予单位】：中国人民解放军医学院
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R644

【相似文献】