CAJ | 학술논문

目的观察创面愈合过程中血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)及其受体表达的动态变化,以及这种变化与创面愈合过程中细胞增殖和凋亡活动之间的关系,探讨AngⅡ在创面愈合过程中的可能作用.方法建立小鼠背部全层皮肤缺损创面模型,于创面形成后第0、1、3、5、7、9、11、13、15天切取创面组织标本,用ELISA法检测创面局部组织AngⅡ产生的变化;采用BrdU及TUNEL染色检测创面细胞增殖和凋亡的变化;用免疫组织化学染色和RT-PCR检测创面局部组织AngⅡ受体AT1和AT2表达的组织细胞定位和mRNA水平的变化.结果小鼠全层皮肤缺损创面局部组织AngⅡ、BrdU标记指数均在伤后逐渐增加,并于第7天达到峰值后即逐渐下降.TUNEL染色阳性细胞数在伤后即开始缓慢增加,并于创面完成上皮化后增加趋势更为明显.正常小鼠皮肤AT1和AT2受体在整个表皮层均有阳性表达,但在真皮层,AT1和AT2受体仅在微血管内皮细胞有阳性表达.AT1受体在角质形成细胞、成纤维细胞均有表达,阳性染色信号在伤后逐渐增加,在第7天最强,以后逐渐下降.AT2受体阳性染色信号也在伤后逐渐增加,7 d以后则逐渐下降.但当创面上皮化完成后,AT2受体阳性染色信号再次增加.RT-PCR结果显示:AT1和AT2受体mRNA均有表达,AT1、AT2受体mRNA表达在伤后第7天均达到峰值,此后则逐渐下降,且AT2受体mRNA表达在创面上皮化完成后表达再次增加.结论在创面愈合过程中,AngⅡ可能通过其产生及受体表达的变化调控创面的愈合及后期的塑形改建.AT1受体可能与细胞增殖活动密切相关;AT2受体可能与细胞凋亡及愈合过程中组织重建有关.
목적 관찰창면유합과정중혈관긴장소Ⅱ(angiotensinⅡ,AngⅡ)급기수체표체적동태변화,이급저충변화여창면유합과정중세포증식화조망활동지간적관계,탐토AngⅡ재창면유합과정중적가능작용.방법 건립소서배부전층피부결손창면모형,우창면형성후제0、1、3、5、7、9、11、13、15천절취창면조직표본,용ELISA법검측창면국부조직AngⅡ산생적변화;채용BrdU급TUNEL염색검측창면세포증식화조망적변화;용면역조직화학염색화RT-PCR검측창면국부조직AngⅡ수체AT1화AT2표체적조직세포정위화mRNA수평적변화.결과 소서전층피부결손창면국부조직AngⅡ、BrdU표기지수균재상후축점증가,병우제7천체도봉치후즉축점하강.TUNEL염색양성세포수재상후즉개시완만증가,병우창면완성상피화후증가추세경위명현.정상소서피부AT1화AT2수체재정개표피층균유양성표체,단재진피층,AT1화AT2수체부재미혈관내피세포유양성표체.AT1수체재각질형성세포、성섬유세포균유표체,양성염색신호재상후축점증가,재제7천최강,이후축점하강.AT2수체양성염색신호야재상후축점증가,7 d이후칙축점하강.단당창면상피화완성후,AT2수체양성염색신호재차증가.RT-PCR결과 현시:AT1화AT2수체mRNA균유표체,AT1、AT2수체mRNA표체재상후제7천균체도봉치,차후칙축점하강,차AT2수체mRNA표체재창면상피화완성후표체재차증가.결론 재창면유합과정중,AngⅡ가능통과기산생급수체표체적변화조공창면적유합급후기적소형개건.AT1수체가능여세포증식활동밀절상관;AT2수체가능여세포조망급유합과정중조직중건유관.
Objective This study was undertaken to observe the change in the local level of angiotensin Ⅱ (Ang Ⅱ) and the expression of its corresponding receptors AT1 and AT2 during wound healing, and explore the possible role of Ang Ⅱ in wound healing . Methods A model of full-thickness cutaneous wound was developed on the back of C57/BL6 mice. Specimens were taken from the wound of each mouse on the day 0, 1, 3, 5, 7, 9, 11, 13 and 15 after wounding. The change in the generation of Ang Ⅱ in wounded tissue during the healing process was detected with ELISA. The proliferation and the apoptosis of cells were detected by bromodeoxyuridine (Brdu) and terminal deoxyuncleotidyl transferase mediated deoxyuridine triphosphate nick end labeling (TUNEL) method in wounded skin during the healing process, respectively. The cellular localization and the mRNA level change of Ang Ⅱ receptors in wounded tissue during healing were detected with immunostaining and RT-PCR. Results Ang Ⅱ produced in wounded skin was increased in the first 7 days to reach the peak, and then gradually decreased during wound healing. BrdU labeling index was increased gradually in the first 7 days to reach the peak, and then gradually decreased during wound healing. The number of TUNEL-positive cells was increased slowly in the first 7 days after wounding. The increase in the number of TUNEL-positive cells was more markedly after epithelization of the wound. In normal mice, AT1 and AT2 receptor were found positively expressed in the whole epidermal layer, while positive expression was only found in the endothelial cells of the capillary vessels within the dermal layer, and positive expression was also found in appendages of the skin, i.e. hair follicle, sweat gland and sebaceous gland respectively. Positive staining signal of both AT1 and AT2 receptors were increased in the first 7 days to reach the peak, then gradually decreased. Expression of AT2R was increased again following the epithelization of wound. The result of RT-PCR showed that the expression of both AT1 and AT2 receptors was detectable, and AT1 receptor was increased in the first 7 days to the peak, and then gradually decreased during wound healing, while AT2 receptor expression reached its peak value on day 7, then gradually decreased, and increased again following the epithelization of wound. Conclusions These results indicate that Ang Ⅱ participate in wound repair and related to remolding in the late stage of wound healing through the change in production of angiotensin Ⅱ and expression of AT1 and AT2 receptors. AT1 receptor might be closely associated with cell proliferation,while AT2 receptor might play a role in cell apoptosis and remolding during wound healing.