Objective To investigate the expression of early growth response protein-1 （Egr-1） in the retina of myopic mice induced by a low-frequency flickering light （FL） and its underlying regulation mechanism in the extracellular matrix remodeling pathway. Methods C57BL/6 mice （n=100） 28 days of age were randomly assigned to two groups： a normal control （n=50） and an FL stimulation group （n=50）. The FL group were raised under illumination with a duty cycle of 50% at a flash rate of 2 Hz. Refractive state and axial length （AL） of the right eyes were measured by a murine-specific eccentric infrared photorefraction and A-scan ultrasonography， respectively. The data were collected at pre-treatment and at 1 hour， 1 day， 1 week， 2 weeks， and after 1 week of recovery. The FL group were removed after 2 weeks of FL stimulation， and the mice were returned to normal light conditions as in the control group. Retinal tissue was collected at each time point to measure the levels of mRNA in Egr-1， membrane type 1 matrix metalloproteinase （MT1-MMP）， matrix metalloproteinase-2 （MMP-2）， and the tissue inhibitor of metalloproteinase-2 （TIMP-2） by quantitative real-time PCR. In addition， the level and location of Egr-1 and MT1-MMP proteins were analyzed by Western Blot， immunohistochemistry and immunofluorescence. A one-way analysis of variance was used to compare the data from the FL and control groups. An independent samples t test was used to compare indexes between the FL group and control group. Results After 2 weeks of FL stimulation， refraction became more myopic compared with the control group （0.32±0.14 D vs. 3.42±0.31 D， t=29.08， P<0.01）， and AL increased faster （2.97±0.01 mm vs. 2.93±0.01 mm， t=6.914， P<0.01）. During the FL treatment phase， Egr-1 mRNA and the protein levels in the treated eyes were rapidly down-regulated after 1 hour， and persistently down-regulated in the retina during treatment. MT1-MMP and MMP-2 levels were up-regulated while TIMP-2 levels were down-regulated in the treated eyes only at time points of 1 or 2 weeks of treatment. During the recovery phase， after 1 week， Egr-1 and TIMP-2 were up-regulated， while MT1-MMP and MMP-2 were down-regulated. Egr-1 and MT1-MMP or MMP-2 were negatively correlated. Immunofluorescence double staining showed Egr-1 immunoreactivity co-localization with MT1-MMP in retinal ganglion cells. Conclusion Retinal Egr-1 may have an effect on the activation of MMP-2 by regulating the expression of MT1-MMP in FL-induced mice myopia and during the recovery process.
李曼,俞莹,管怀进,陈辉. 视网膜Egr-1与细胞外基质重塑通路对闪烁光诱导小鼠近视的调控[J]. 中华眼视光学与视觉科学杂志, 2014, 16(6): 328-334.
Li Man,Yu Ying,Guan Huaijin,Chen Hui. The modulation of retinal Egr-1 and the extracellular matrix remodeling pathway in flickering light-induced myopia in mice. Chinese Journal of Optometry Ophthalmology and Visual Science, 2014, 16(6): 328-334. DOI: 10.3760/cma.j.issn.1674-845X.2014.06.003
Crewther SG， Barutchu A， Murphy MJ， et al. Low frequency temporal modulation of light promotes a myopic shift in refractive compensation to all spectacle lenses[J]. Exp Eye Res，2006，83：322-328.
[2]
Yu Y， Chen H， Tuo J， et al. Effects of flickering light on refraction and changes in eye axial length of c57bl/6 mice[J]. Ophthalmic Res，2011，46：80-87.
Zhou X， An J， Wu X， et al. Relative axial myopia induced by prolonged light exprosure in C57BL/6mice[J]. Photochem Photobiol，2010，86：131-137.
[7]
McBrien NA， Jobling AI， Gentle A. Biomechanics of the sclera in myopia： Extracellular and cellular factors[J]. Optom Vis Sci，2009，86：E23-30.
[8]
Wallman J. Retinal control of eye growth and refraction[J]. Prog Retinal Res，1993，12：133-153.
[9]
Fischer AJ， McGuire JJ， Schaeffel F， et al. Light- and focus-dependent expression of the transcription factor zenk in the chick retina[J]. Nat Neurosci，1999，2：706-712.
[10]
Schippert R， Burkhardt E， Feldkaemper M， et al. Relative axial myopia in Egr-1 （ZENK） knockout mice[J]. Invest Ophthalmol Vis Sci，2007，48：11-17.
Ashby R， McCarthy CS， Maleszka R， et al. A muscarinic cholinergic antagonist and a dopamine agonist rapidly increase ZENK mRNA expression in the form-deprived chicken retina[J]. Exp Eye Res，2007，85：15-22.
[13]
Liu C， Adamson E， Mercola D. Transcription factor EGR-1 suppresses the growth and transformation of human HT-1080 fibrosarcoma cells by induction of transforming growth factor beta1[J]. Proc Natl Acad Sci USA，1996，93：11831-11836.
[14]
Siegwart JT Jr， Norton TT. Selective regulation of mmp and timp mRNA levels in tree shrew sclera during minus lens compensation and recovery[J]. Invest Ophthalmol Vis Sci，2005， 46：3484-3492.
[15]
Haas TL， Stitelman D， Davis SJ， et al. Egr-1 mediates extracellular matrix-driven transcription of membrane type 1 matrix metalloproteinase in endothelium[J]. J Biol Chem，1999， 274：22679-22685.
[16]
Xiang F， Bai M， Jin Y， et al. Egr-1 mediates si0（2）-driven transcription of membrane type i matrix metalloproteinase in macrophages[J]. J Huazhong Univ Sci Technolog Med Sci，2007， 27：13-16.
[17]
Ning W， Dong Y， Sun J，et al. Cigarette smoke stimulates matrix metalloproteinase-2 activity via Egr-1 in human lung fibroblasts[J]. Am J Respir Cell Mol Biol，2007，36：480-490.
[18]
Guggenheim JA， McBrien NA. Form-deprivation myopia induces activation of scleral matrix metalloproteinase-2 in tree shrew[J]. Invest Ophthalmol Vis Sci，1996，37：1380-1395.
[19]
Siegwart JT Jr， Strang CE. Selective modulation of scleral proteoglycan mRNA levels during minus lens compensation and recovery[J]. Mol Vis，2007，13：1878-1886.
[20]
Karagiannis ED， Popel AS. A theoretical model of type i collagen proteolysis by matrix metalloproteinase （mmp） 2 and membrane type 1 mmp in the presence of tissue inhibitor of metalloproteinase 2[J]. J Biol Chem，2004，279：39105-39114.
[21]
Schaeffel F， Burkhardt E， Howland HC， et al. Measurement of refractive state and deprivation myopia in two strains of mice[J]. Optom Vis Sci，2004，81：99-110.
[22]
Agapova OA， Kaufman PL， Lucarelli MJ， et al. Differential expression of matrix metalloproteinases in monkey eyes with experimental glaucoma or optic nerve transection[J]. Brain Res， 2003，967：132-143.
Caputto BL， Guido ME. Immediate early gene expression within the visual system： Light and circadian regulation in the retina and the suprachiasmatic nucleus[J]. Neurochem Res，2000，25：153-162.
[26]
Ashby R， Kozulin P， Megaw PL， et al. Alterations in zenk and glucagon rna transcript expression during increased ocular growth in chickens[J]. Mol Vis，2010，16：639-649.
浙公网安备 33030202001892号