Comprehensive analysis of differentially expressed genes responsed to mechanical stretch in human corneal keratocytes.. Comprehensive analysis of differentially expressed genes responsed to mechanical stretch in human corneal keratocytes.

Mechanical stimulation is an important factor for the development of complications after LASIK, but the molecular mechanism still remains unclear. In this study, we have employed whole genome microarray expression profiling as a discovery platform to identify genes responded to mechanical stretch in human corneal keratocytes. Furthermore, the specific signal transduction pathways were discussed. Mechanical stretch regulated gene expression was measured in human corneal keratocytes subjected to cyclic stretch (0-15% elongation, 0.5 Hz, sine waveform) for 0, 1 and 6 hours, respectively. Totally, 840 differentially expressed genes were identified in keratocytes under mechanical stretching, among which 493 genes were up-regulated and 388 genes were down-regulated. Theses differential genes were mainly involved in cytokine-cytokine receptor interaction, ECM-receptor interaction, Focal adhesion, TNF signaling pathway, and MAPK signaling pathway. Overall design: Mechanical stretch regulated gene expression was measured in human corneal keratocytes subjected to cyclic stretch (0-15% elongation, 0.5 Hz, sine waveform) for 0, 1 and 6 hours, respectively. Three independent experiments were performed for each stretch time (0, 1 or 6 hours).

机械刺激是准分子激光原位角膜磨镶术（LASIK）术后并发症发生的重要影响因素，但其具体分子机制尚未明确。本研究采用全基因组微阵列表达谱作为筛选平台，旨在鉴定人角膜基质细胞（human corneal keratocytes）中响应机械牵拉的基因，并对相关特异性信号转导通路进行了探讨。
本研究分别对施加0、1、6小时周期性机械牵拉（0-15%伸长率、0.5 Hz、正弦波形）的人角膜基质细胞的机械牵拉调控基因表达情况进行了检测。最终在机械牵拉处理的角膜基质细胞中鉴定出840个差异表达基因（differentially expressed genes），其中493个基因表达上调，388个基因表达下调。
上述差异表达基因主要富集于细胞因子-细胞因子受体相互作用、细胞外基质（Extracellular Matrix, ECM）-受体相互作用、黏着斑（Focal adhesion）、肿瘤坏死因子（Tumor Necrosis Factor, TNF）信号通路以及丝裂原活化蛋白激酶（Mitogen-Activated Protein Kinase, MAPK）信号通路中。
实验设计：本研究分别对施加0、1、6小时周期性机械牵拉（0-15%伸长率、0.5 Hz、正弦波形）的人角膜基质细胞的机械牵拉调控基因表达情况进行检测，每个牵拉时间点（0、1、6小时）均开展3次独立重复实验。