细胞外基质金属蛋白酶及其抑制剂(EMMPs/TIMPs)在玻璃体细胞介导胶原凝胶收缩中的调控

This study is designed to explore the regulative mechanism of matrix metalloproteinases/tissue inhibitors of matrix metalloproteinases (EMMPs/TIMPs) in hylalocytes mediated collagen contraction, and finally to find a new therapy for early prevention of contractional vitreoretinal diseases. In vitro cell mediated collagen contraction model is used in the study, the model is prepared on ice and contained collagen type II mixture and hyalocytes, the mRNA and protein expression levels of EMMPs/TIMPs in this hyalocytes are detected by real-time PCR and ELSA, respectively. The activities of secreted EMMPs/TIMPs in the model are analyzed by gelatin/casein zymography and reserved zymography. Following groups are established in the study: exogenous EMMPs/TIMPs addition groups and control group. Before and after collagen gel contraction, the gel volume and diameter, water/gel proportion and 3D-structure of the gel are all recorded in each group, to analyze the effect EMMPs/TIMPs on the hyalocytes mediated collagen gel contraction.

玻璃体视网膜界面性疾病(VRID)是当前备受关注的致盲眼病，玻璃体与视网膜的粘连收缩是主要致病因素。我们前期研究已证实玻璃体细胞是玻璃体胶原凝胶三维构像维持和收缩性状变化的重要细胞介质，玻璃体胶原的异常铰链和三维空间构像塌陷，是玻璃体收缩的内在分子学表像和特征，但其调控途径不甚明确。最新研究发现细胞外基质金属蛋白酶及其抑制剂（EMMPs∕TIMPs）是细胞介导胶原收缩的重要启动子，本课题旨在从EMMPs∕TIMPs途径揭示玻璃体细胞介导胶原凝胶收缩的上游调控机制。建立细胞介导胶原凝胶收缩模型，通过胶原凝胶柱直径、体积、水∕凝胶比重及扫描电镜三维构像变化，通过外源性EMMPs和TIMPs的调控，对比各亚型EMMPs∕TIMPs对玻璃体细胞介导胶原收缩的影响，旨在拓展对VRID上游EMMPs∕TIMPs调控的新认识，探索早期干预途径。

玻璃体视网膜界面性疾病是当前备受关注的致盲眼病，玻璃体与视网膜的粘连收缩是主要致病因素。我们前期研究已证实玻璃体胶原凝胶支架结构的异常铰链和空间构象的改变是导致玻璃体胶原异常收缩的内在机制，玻璃体细胞可能是收缩产生的动力来源，但其调控途径仍不明确。本课题通过细胞外基质金属蛋白酶及其抑制剂（EMMPs/TIMPs）为切入点，应用体外凝胶模型构建、扫描电镜、Realtime-PCR、ELISA、明胶酶谱等技术和方法深入研究发现：（1）体外构建的玻璃体细胞介导的胶原凝胶收缩模型中可检测出多种EMMPs及TIMPs的表达。（2）玻璃体细胞介导的胶原凝胶收缩模型随着细胞培养时间的延长而发生收缩，收缩程度与模型中的EMMPs含量呈正相关。（3）外源性添加EMMPs可显著加速玻璃体细胞介导的胶原凝胶模型的收缩，而外源性添加TIMPs可缓解收缩的发生。