pVHL/HIF-1信号通路在滤过术后瘢痕化双向调控中的作用及机制研究

Glaucoma is the leading cause of global irreversible blindiness. Filtration surgery is one of the main metholds of treatment, while postoperative wound healing and scarring processes are important factors affecting postoperative prognosis. Both excessive and insufficient scar formation are major causes of surgical failure after glaucoma filtration surgery. Scarring regulation is not only the focus of current research but also the difficult problem of glaucoma urgent to be solved. Based on the previous work, the project plan to up and down regulate hypoxia-inducible factor (HIF-1)-mediated hypoxia-inducible pathways through HIF-1 upstream regulatory factors pVHL to further explore the balanced control mechanisms of pVHL/HIF-1 pathway in the regulation of scarring regulated by hypoxia-inducible pathway. The study use Tenon's broblasts, conjunctiva epithelial cells cultured in hypoxic conditions and the rabbit filtering surgery model. By using si-RNA, gene transduction, Western blot and PCR technology, observe changes in the downstream pathway molecules during hypoxia and scar formation. The results will be helpful to reveal the molecular mechanism of pVHL/HIF-1 signal pathway in balanced control of scar formation after filtering surgery and is expected to provide a new effective, low toxic, low side effect and bidirectional regulation intervention method of postoperative scar after filtering surgery.

青光眼是全球首位不可逆性致盲眼病。滤过手术是其治疗的主要途径之一，滤过术后创口愈合及瘢痕化过程是影响手术预后的重要因素，瘢痕化过度及不足均是手术失败的主要原因。瘢痕化调控既是目前的研究热点，也是青光眼亟待解决的难题。项目在前期工作基础上拟利用缺氧诱导因子（HIF-1）上游调控因子pVHL上调及下调HIF-1介导的缺氧诱导通路，进一步探讨pVHL/HIF-1调控的缺氧诱导通路在滤过术后瘢痕化过程中的双向调控作用机制。以缺氧条件下培养的Tenon's囊成纤维细胞、结膜上皮细胞及滤过术后瘢痕化兔为模型，运用si-RNA、基因转导及Western blot等技术，研究缺氧和创口愈合及瘢痕化发生发展过程中HIF-1介导的缺氧诱导通路及其上下游分子的变化。旨在揭示pVHL/HIF-1信号通路在滤过术后瘢痕化过程中双向调控的分子机制，有望为滤过术后瘢痕化过程的双向调控提供既有效又低毒低副作用的临床路径。

摘要.青光眼是全球首位不可逆性致盲眼病。滤过手术是其治疗的主要途径之一，滤过术后创口愈合及瘢痕化过程是影响手术预后的重要因素，瘢痕化过度及不足均是手术失败的主要原因。瘢痕化调控既是目前的研究热点，也是青光眼亟待解决的难题。项目通过构建人Tenon's囊成纤维细胞模型，发现缺氧诱导因子-1(Hypoxia inducible factor)HIF-1在人Tenon’s囊成纤维细胞组织存在表达。进一步研究发现HIF-1过表达可能促进结膜瘢痕化。通过构建兔结膜伤口愈合模型，检测在兔结膜伤口的表达水平，结果发现,HIF-1α在兔结膜和Tenon’s囊组织存在表达.HIF-1过表达可能促进兔结膜伤口愈合。通过VH-298及KC7F2分别上、下调HTFs中HIF-1的表达，观察到HIF-1对其下游瘢痕相关因子VEGF有正向调节作用。同时检测到CollagenⅠ在HIF-1上调时增加表达，在HIF-1下调时减少表达。研究结果认为HIF-1是滤过术后瘢痕化形成过程中的重要靶标因子，为滤过术后瘢痕化过程的双向调控提供了既有效又低毒低副作用的临床路径。