| 目的 探讨外源性RA对小鸡后极部巩膜基质金属蛋白酶2（MMP-2）mRNA及蛋白质表达水平的影响。材料与方法 取1d龄来亨雏鸡，右眼球后注射RA，制备外源性RA实验组标本，对侧眼为自身对照组，阴性对照组右眼球后注射生理盐水，正常对照组小鸡不做任何处理。分别于注射后12h、24h、48h、72h摘取眼球，采用一步法逆转录－多聚酶链反应（RT-PCR法）及免疫组化法检测每组小鸡后极部巩膜MMP-2mRNA和蛋白质表达水平。结果 与正常组、阴性对照组相比，实验组后极部巩膜MMP-2 mRNA表达显著增高，组间差异有显著性（P﹤0.01）。注射后随时间延长MMP-2 mRNA表达逐渐上调，48h达最高峰，至72h有降低。不同时间组组间差异显著（P﹤0.01）。自身对照组MMP-2 mRNA表达较同龄正常对照组有轻度上调，组间有统计学差异（P﹤0.01）。免疫组化显示，实验组MMP-2蛋白质表达水平增高，主要在巩膜成纤维细胞层。结论 外源性RA能够上调巩膜MMP-2mRNA及蛋白质水平，可能是巩膜细胞外基质主动重塑的启动因子。
关键词 视黄酸，基质金属蛋白酶2 ，基质金属蛋白酶组织抑制剂，巩膜重塑
The Effect of Retinoic Acid in Changes of MMP-2 and TIMP-2 Expression in the Posterior Sclera of Chick eye
Wang jianfeng, Liu shuangzhen, Wu wencan, Department of Ophthalmology, Xiangya Hospital in Central South University, Changsha 410008,
【Abstract】Objective To investigate the changes of matrix metalloproteinase 2(MMP-2) mRNA expression and protein level in posterior sclera of chick eye after a injection of RA. Methods1-day-old white leghorn chicks were divided randomly and equivalently into three groups, RA was retrobulbar injected to the right eye of each chick to induce the experimental group, and the left eye served as self-control. NS injection induced the negative control group. Meanwhile, normal age-matched animals were provided as normal control group. Eyes were collected at 12, 24, 48 and 72 hour after injection, and expression levels of MMP-2 mRNA were analyzed by single step reverse transcription-polymerase chain reaction (RT-PCR). The protein level of MMP-2 was analyzed by immunohistochemistry as well. Results Compared with normal and self-controlled groups, the level of MMP-2 mRNA expression in the posterior sclera of the experimental group was high. There was statistical difference between experimental groups and control groups(P﹤0.01); As the time passed, levels of MMP-2 mRNA expression increased significantly and reached a peak at 48hour after injection, and then decreased slightly but still maintained in a relative high level. There was statistical differency between groups in different times(P﹤0.01). There was statistical differency between normal and self-controlled groups (P<0.01), seemed as the RA injected effected by topediffusion. Immunohistochemistry showed that the expression of MMP-2 was mainly changed in the fibroblast cell layer of sclera. Conclusion Retinoic acid can effect on the expression of MMP-2 in the Posterior segment of eye directly or indirectly. RA may be an important factor of extracellular matrix (ECM) remodeling in sclera.
Key words Retinoic Acid; Matrix metalloproteinase-2；Tissue inhibitor of matrix metalloproteinase-2；Extracellular matrix remodeling, Sclera. |
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