低氧诱导因子在鼻息肉中的表达与功能

发布时间：2018-10-21 10:14

【摘要】：研究背景和目的慢性鼻-鼻窦炎(chronic rhinosinusitis,CRS)是一组以鼻腔鼻窦黏膜的慢性炎症为特征的常见疾病,慢性鼻-鼻窦炎伴有鼻息肉(chronic rhinosinusitis with nasal polyps,CRSwNP)就是其中病情较顽固、临床上较为难治的类型。CRSwNP的主要病理特征为鼻黏膜水肿、炎症细胞浸润、上皮异常增生。低氧一直被认为是参与CRSwNP发病的因素之一,但其中的机制尚不明确。在低氧的情况下,组织细胞会激活一系列的调节机制,通过减少氧气消耗、促进血管生成和改变代谢途径等方式来适应低氧的环境,这其中有许多过程是受到低氧诱导因子(hypoxia-inducible factor,HIF)直接或间接的调控的。HIF是一类转录因子,在氧含量下降时,可以被激活,进而对多种基因的表达水平进行调控,在细胞适应低氧环境的过程中起到重要作用。本课题选择对HIF-1α、HIF-2α及HIF-3α在CRSwNP中的表达与功能进行进一步研究,为CRSwNP的临床治疗与研究提供一定的理论基础。方法从鼻内镜手术中获取鼻息肉组织40例为鼻息肉组,所有样本均经病理检查证实为鼻息肉组织。20例行鼻内镜手术的鼻中隔偏曲患者的下鼻甲黏膜作为对照组,对照组患者均无慢性鼻-鼻窦炎、鼻息肉、哮喘、过敏性鼻炎或其他系统性疾病。术中取得的新鲜标本被分为两部分,一部分固定于4%甲醛固定液中,常规石蜡包埋,以供HE染色、免疫组织化学染色及免疫荧光染色,进行组织学评估;使用RNA later保存另一部分组织,以供后续进行提取组织mRNA并分析基因表达的研究。统计分析应用SPSS 22.0软件,使用Mann-Whtiney检验比较鼻息肉组和对照组中目的基因的蛋白和mRNA表达水平,应用Spearman相关分析对所选基因的相关关系进行评价,p0.05表示差异有统计学意义。结果1.与对照组相比,鼻息肉组HIF-2α的蛋白和mRNA表达水平均有显著的降低,差异有统计学意义(p0.01);HIF-3α的mRNA表达水平有显著的降低(p0.01),蛋白的表达差异无统计学意义;HIF-1α的mRNA及蛋白表达差异均无统计学意义。2.HIF-1α、HIF-2α及HIF-3α在组织中的表达部位存在差异,HIF-1α和HIF-3α主要表达在上皮内,HIF-2α主要表达在上皮下,腺体和血管周围均有表达。3.与对照组相比,鼻息肉组VE-钙粘蛋白的蛋白和mRNA表达水平均有显著的降低,差异有统计学意义(p0.01),HIF-2α可能参与了这一变化。结论在鼻息肉中,三种HIF-α亚型的表达水平和表达部位存在差异,可能在鼻息肉的发病过程中起不同的作用,对HIF-α特别是HIF-2α及其相关因子的深入研究,可能成为潜在的靶向治疗的治疗靶点。
[Abstract]:Background and objective chronic rhinosinusitis (chronic rhinosinusitis,CRS) is a common disease characterized by chronic inflammation of nasal cavity and sinusitis mucosa. Chronic rhinosinusitis with nasal polyp (chronic rhinosinusitis with nasal polyps,CRSwNP) is one of the most obstinate diseases. The main pathological features of CRSwNP are edema of nasal mucosa, infiltration of inflammatory cells and abnormal hyperplasia of epithelium. Hypoxia has been considered to be one of the factors involved in the pathogenesis of CRSwNP, but the mechanism is unclear. In hypoxic conditions, tissue cells activate a series of regulatory mechanisms to adapt to hypoxia by reducing oxygen consumption, promoting angiogenesis, and altering metabolic pathways. Many of these processes are directly or indirectly regulated by hypoxic inducible factors (hypoxia-inducible factor,HIF). HIF is a class of transcription factors that can be activated when oxygen levels decrease, thereby regulating the expression levels of multiple genes. It plays an important role in the process of cells adapting to hypoxic environment. In this paper, we choose to further study the expression and function of HIF-1 伪, HIF-2 伪 and HIF-3 伪 in CRSwNP, and provide a theoretical basis for the clinical treatment and study of CRSwNP. Methods 40 cases of nasal polyps were obtained from endoscopic sinus surgery. All samples were proved to be nasal polyps by pathological examination. 20 cases of nasal septum deviation were used as control group. No chronic rhinosinusitis, nasal polyps, asthma, allergic rhinitis or other systemic diseases were found in the control group. The fresh specimens were divided into two parts, one was fixed in 4% formaldehyde fixed solution, and the other was embedded in paraffin for HE staining, immunohistochemical staining and immunofluorescence staining for histological evaluation. Another part of tissue was preserved with RNA later for further study of mRNA extraction and gene expression analysis. The expression levels of protein and mRNA of the target gene in nasal polyp group and control group were compared by using SPSS 22.0 software and Mann-Whtiney test. The correlation of the selected gene was evaluated by Spearman correlation analysis. The difference was statistically significant. Result 1. Compared with the control group, the expression of HIF-2 伪 protein and mRNA in nasal polyp group were significantly decreased. The difference was significant (p0.01), the mRNA expression of HIF-3 伪 was significantly decreased (p0.01), the expression of mRNA and protein of HIF-1 伪 was not significantly different, the expression of 2.HIF-1 伪, HIF-2 伪 and HIF-3 伪 were different in the tissues, HIF-1 伪 and HIF-3 伪 were different in the tissues, HIF-1 伪 and HIF-3 伪 had no significant difference in the expression of 2.HIF-1 伪, HIF-2 伪 and HIF-3 伪. HIF-3 伪 was mainly expressed in the epithelium, and HIF-2 伪 was mainly expressed in the subepithelium. Gland and perivascular expression. 3. 3. Compared with the control group, the expression of VE- cadherin protein and mRNA in nasal polyp group was significantly lower than that in control group (p0.01). HIF-2 伪 may be involved in this change. Conclusion in nasal polyps, there are differences in the expression levels and expression sites of the three HIF- 伪 subtypes, which may play different roles in the pathogenesis of nasal polyps. The study of HIF- 伪, especially HIF-2 伪 and its related factors, may play a different role in the pathogenesis of nasal polyps. It may be a potential therapeutic target for targeted therapy.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R765

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