Hippo信号通路在病理性瘢痕发生机制中作用研究

发布时间：2018-05-13 04:27

本文选题：病理性瘢痕 + Hippo信号通路　；参考：《扬州大学》2017年硕士论文

【摘要】：研究背景:瘢痕是临床上常见的疾病之一,可分为病理性瘢痕和生理性瘢痕两类。瘢痕常形成于创伤后,适度的瘢痕形成是一种生理性和自卫性的表现,但瘢痕过度增生则属于病理性的表现。病理性瘢痕一般可分为增生性瘢痕和瘢痕疙瘩,常常对患者的外观形象以及瘢痕部位的活动功能等产生不良影响,同时也会造成患者的心理负担,严重者可导致抑郁症等疾病发生。现代社会随着经济的发展和人们生活水平的提高,患者对于生活质量的要求以及美容方面的要求越来越高,因此对于病理性瘢痕的病因探索、诊断治疗新技术等的需求也更为强烈。病理性瘢痕的两大特征为成纤维细胞的过度增殖与瘢痕的类肿瘤性特征。目前研究表明成纤维细胞是创伤后创面愈合的主要修复细胞,在创面修复的过程中活化、增殖、合成胶原,其过度增殖是病理性瘢痕的一个主要特征。病理性瘢痕在病因、病理机制、临床表现、诊断和治疗等方面与肿瘤均具有相似的生物学特性,即病理性瘢痕的类肿瘤性特性。病理性瘢痕的类肿瘤性特性一方面能够较好、较全面地概述病理性瘢痕的生物学特性,另一方面强调了病理性瘢痕防治的复杂性和艰巨性,因此对于病理性瘢痕发生机制研究可借鉴肿瘤学的研究思路和成果。2005年,有学者首次在果蝇中发现Hippo信号通路传递从细胞质到细胞核的细胞信号,该通路能够调节下游基因的表达,从而调节细胞的生长、增殖和凋亡,参与调控器官大小和组织再生,在胚胎发育和组织器官形成等过程中具有重要作用。研究表明Hippo信号通路参与调解多种成纤维细胞的形态功能,同时也与肿瘤的发生、发展有直接的关系。综上所述,我们猜测Hippo信号通路在病理性瘢痕形成过程中发挥一定的作用。目前已知YAP/TAZ的表达在Hippo信号通路中发挥关键作用,因此本研究拟通过实验阐述Hippo信号通路关键性作用分子YAP/TAZ在病理性瘢痕中表达情况,探索Hippo信号通路在病理性瘢痕发生机制中的作用。目的:研究人真皮成纤维细胞及人皮肤增生性瘢痕成纤维细胞中Hippo信号通路关键作用分子YAP/TAZ表达情况,并初步建立增生性瘢痕兔耳模型,检测兔耳增生性瘢痕中YAP/TAZ表达情况。方法:(1)流式细胞术检测人真皮成纤维细胞及人皮肤增生性瘢痕成纤维细胞凋亡情况,免疫印迹试验(Western Blot)、逆转录-聚合酶链反应(Reverse transcriotion-polymerase chain reaction,RT-PCR)、定量实时聚合酶链反应(Quantitative real time polymerase chain reaction,Q-PCR)方法检测人真皮成纤维细胞及人皮肤增生性瘢痕成纤维细胞中Hippo信号通路主要作用分子YAP/TAZ表达情况。(2)兔耳瘢痕模型建立:用环钻在每只兔耳腹侧面各做6个直径6mm的圆形全层皮肤缺损创面,每个创面间隔2cm以上,去除软骨膜,创伤后约21天时取瘢痕组织做苏木素一伊红(Hematoxylin and eosin,HE)染色检测瘢痕形成情况,用Western Blot方法检测YAP/TAZ蛋白表达情况。结果:(1)人皮肤增生性瘢痕成纤维细胞凋亡弱于人真皮成纤维细胞,YAP/TAZ表达人皮肤增生性瘢痕成纤维细胞高于人真皮成纤维细胞。(2)兔耳创伤后约21天左右形成增生性瘢痕,HE染色显示胶原增生,成纤维细胞增生,瘢痕组织YAP/TAZ表达高于正常兔耳组织。结论:Hippo信号通路特别是其关键作用分子YAP/TAZ在病理性瘢痕发生过程中起到一定的作用。
[Abstract]:Research background: scar is one of the common diseases in clinical. It can be divided into two types: pathological scar and physiological scar. Scar is often formed after trauma. Moderate scar formation is a physiological and self-defense manifestation, but hypertrophic scar is a pathological manifestation. Pathological cicatrix can be divided into hypertrophic scar and kelp. In one's heart, it often has a bad effect on the appearance of the patient and the activity function of the scar. It can also cause the psychological burden of the patient, and the serious person can cause the disease of depression. In modern society, with the development of the economy and the improvement of people's living standard, the more the patients demand for the quality of life and the beauty. The higher it is, the need for the pathogeny exploration of pathological scar and the new technique of diagnosis and treatment are also stronger. The two characteristics of pathological scar are the hyperproliferation of fibroblasts and the tumor like characteristics of scar. Activation, proliferation, and collagen synthesis, and their excessive proliferation is a major feature of pathological scars. Pathological scars have similar biological characteristics with tumors in the etiology, pathological mechanism, clinical manifestations, diagnosis and treatment, that is, the tumor like characteristics of the cicatricial cicatrix. The tumor like characteristics of pathological scars can be compared on one hand. Well, a comprehensive overview of the biological characteristics of pathological cicatrix is outlined. On the other hand, the complexity and arduous nature of the prevention and control of pathological scars is emphasized. Therefore, for the study of the pathogenesis of pathological cicatrix, the study of oncology can be used for reference in.2005 years. Some scholars first found the Hippo signaling pathway from the cytoplasm to the nucleus in the Drosophila. Cell signaling, which regulates the expression of downstream genes, regulates cell growth, proliferation and apoptosis, participates in the regulation of organ size and tissue regeneration, and plays an important role in the process of embryonic development and tissue formation. The study shows that Hippo signaling pathway participates in the mediation of the morphological functions of multiple fibroblasts, as well as in the form of fibroblasts. There is a direct relationship with the development of the tumor. In summary, we have guessed that the Hippo signaling pathway plays a role in the formation of pathological scar. The expression of YAP/TAZ is known to play a key role in the Hippo signaling pathway. Therefore, this study intends to elaborate the key role of the Hippo signaling pathway, YAP/TAZ, in the disease. To explore the role of Hippo signaling pathway in the pathogenesis of pathological scar. Objective: To study the expression of the key role of Hippo signaling pathway in human dermal fibroblasts and human dermal hypertrophic scar fibroblasts, and to establish an initial model of hypertrophic scar rabbit ears to detect the proliferation of rabbit ears. YAP/TAZ expression in the scar. Methods: (1) flow cytometry was used to detect the apoptosis of human dermal fibroblasts and human dermal hypertrophic scar fibroblasts, Western Blot, reverse transcription polymerase chain reaction (Reverse transcriotion-polymerase chain reaction, RT-PCR), and quantitative real-time polymerase chain reaction (Quantitative R). Eal time polymerase chain reaction, Q-PCR) method to detect the main role of Hippo signaling pathway YAP/TAZ expression in human dermal fibroblasts and human dermal hypertrophic scar fibroblasts. (2) the rabbit ear scar model was established: 6 round full layer skin defect wounds with diameter 6mm in each rabbit ear were made by ring drill. The wound interval was above 2cm, and the cartilage membrane was removed. The scar tissue was stained by Hematoxylin and eosin (HE) to detect the scar formation when the scar tissue was stained about 21 days after the trauma. The expression of YAP/TAZ protein was detected by Western Blot. Results: (1) the apoptosis of human dermal proliferative scar fibroblasts was weaker than human dermal fibroblasts, YAP/TAZ The expression of human dermal hypertrophic scar fibroblasts was higher than that of human dermal fibroblasts. (2) hypertrophic scar formed about 21 days after trauma in rabbit ears. HE staining showed collagen proliferation, fibroblast proliferation, and YAP/TAZ expression in scar tissue was higher than normal rabbit ear tissue. Conclusion: the Hippo signal pathway, especially its key molecule YAP/TAZ, is in pathology It plays a certain role in the process of sexual scarring.

【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R622

【参考文献】