[关键词]
[摘要]
目的:观察C57BL/6小鼠形觉剥夺性近视(form-deprivation myopia,FDM)巩膜中BMP-2表达的变化,研究其在巩膜重塑中发挥的作用。
方法:选择3~4周龄的C57BL/6小鼠64只,以随机数字表法随机分为形觉剥夺21d组(16只)、同龄对照组(16只); 形觉剥夺28d组(16只)、同龄对照组(16只)。形觉剥夺前后对所有小鼠进行带状光剪影验光检测小鼠眼球的屈光状态,游标卡尺测量小鼠的眼轴长度,造模后分别于21d和28d取小鼠的巩膜组织,苏木素-伊红(HE)染色观察小鼠巩膜组织形态学变化,用荧光定量RT-PCR和免疫组织化学检测各组小鼠巩膜BMP-2 mRNA和其蛋白的表达水平。
结果:小鼠形觉剥夺21d和28d后,剥夺眼分别诱导出-1.60±1.03D和-3.10±1.19D的相对近视,眼轴分别拉长16±12μm和21±13μm,与自身对照组和正常对照相比较,差异有统计学意义(P<0.05)。HE染色行巩膜形态学观察,可观察到剥夺眼巩膜变薄,胶原纤维排列紊乱。荧光定量RT-PCR和免疫组织化学结果显示实验眼BMP-2 mRNA和蛋白的表达明显下调,与自身对照和正常对照组相比差异有统计学意义(P<0.05)。
结论:C57BL/6小鼠形觉剥夺眼巩膜中BMP-2呈下调趋势,BMP-2可能参与了近视发展过程中的巩膜重塑作用,其具体机制有待进一步研究。
[Key word]
[Abstract]
AIM: To identify the presence change of bone morphogenetic protein-2(BMP-2)in sclera of form deprivation myopic(FDM)eyes in C57BL/6 mice and to investigate its role in sclera remodeling.
METHODS: A total of 64 C57BL/6 mice which were 3~4wk were randomized into FDM-21d group(n=16),a normal control group(n=16); FDM-28d group(n=16)and a normal control group(n=16). FDM models were established by sutured 0.5mL PCR plastic caps to the skin surrounding the right eye for 21 days and 28 days in experimental group, and the fellow eyes served as the self-control eyes. Diopter and axial length of all eyes was tested by retinoscopy optometry and vernier calipers before and after the form deprivation. As the models established, the sclera of the mice were obtained and hematoxylin and eosin(H-E)staining were used for observing the morphological change in the tissue. Immunohistochemical staining and fluorescence quantificational real-time polymerase chain reaction(QRT-PCR)were applied to investigate the expression of BMP-2 protein and mRNA.
RESULTS: After 21 and 28d,the diopter of the deprived eyes was -1.60±1.03D and -3.10±1.19D and the axial length was prolonged by 16±12μm and 21±13μm respectively. The differences were statistically significant, compared to self-control eyes and to normal control groups(P<0.05). After stained by H-E, the sclera from the deprived eyes became thinner and the sequence of collagenous fiber disappeared. The results from the immunohistochemical staining and QRT-PCR showed that mRNA and protein of BMP-2 decreased significantly, and the differences were statistically significant, compared to self-control eyes and to normal control groups(P<0.05).
CONCLUSION: The expressions of the BMP-2 in sclera down-regulate significantly in FDM eyes,which suggests that BMP-2 may play an important role in sclera remodeling during myopia development.
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[基金项目]
国家自然科学基金青年科学基金项目(No.81300790); 山东省自然基金项目(No.BS2010SF009)