Sp1和Ⅰ型胶原在豚鼠实验性近视巩膜重塑中的表达及其相关性研究

发布时间：2018-10-24 12:01

【摘要】：目的:研究近视巩膜重塑中Sp1作为TGF-β1下游信号转录基因的表达情况。探讨Sp1蛋白分子及m RNA的表达与实验性形觉剥夺性近视(form deprived myopia,FDM)之间的关系。以期揭示TGF-β1-Sp1-CollagenⅠ信号途径在近视巩膜胶原重塑中的作用。方法:选取出生1周左右的健康三色豚鼠75只,利用6号半透明乳胶气球作为眼罩遮盖豚鼠的眼睛以诱导形觉剥夺性近视动物模型。将75只豚鼠随机分为正常对照组(50眼)、形觉剥夺(FDM组,50眼)组和自身对照组(50眼),正常对照组是未做任何处理的豚鼠,FDM组是实验组的豚鼠遮盖眼,自身对照组是实验组豚鼠的暴露眼,方法是使用半透明的乳胶气球,依据豚鼠的头部形状制作成头套,遮盖豚鼠的左眼,暴露其右眼、口鼻及双耳。这种方法具有稳定、无害、不会对角膜产生压力和温度变化等诸多的优势。分别测量记录实验豚鼠在未遮盖前(0周)、遮盖2周、4周、6周及遮盖4周去遮盖1周(4/-1周)时的双眼屈光度及眼轴长度。采用免疫组织化学技术、RT-PCR法检测豚鼠正视眼和实验性近视眼巩膜中分别在遮盖0周、2周、4周、6周及诱导4周去诱导1周时间段内SP1的动态表达变化以及与Ⅰ型胶原合成与表达的变化关系。结果:形觉剥夺组(FDM组)由遮盖前(0周)的远视眼(+2.09±0.31)D到遮盖2周(-1.23±0.69)D、4周(-4.17±0.59)D、遮盖4周去遮盖1周(-4.30±0.58)D及遮盖6周(-7.07±0.56)D后逐渐变成近视眼,且随着时间延长,近视度数不断加深,眼轴也逐渐延长[遮盖前眼轴长:(5.93±0.39)mm;2周时眼轴长:(6.62±0.36)mm;4周时眼轴长:(7.30±0.34)mm;4周去1周眼轴长:(7.41±0.36)mm;6周时眼轴长:(7.99±0.32)mm,P0.05]。免疫组化结果显示,形觉剥夺(FDM)组巩膜有Sp1阳性表达,其表达均弱于两组对照组,且随着遮盖时间的延长,Sp1的阳性表达在不断减弱(p0.05)。RT-PCR结果认为,和两组对照组相比,形觉剥夺组Sp1的m RNA表达均下调(p0.05),且随着时间的延长,m RNA的表达也在逐渐下调(p0.05)。结论:Sp1是TGF-β1的下游信号转录基因,且在豚鼠巩膜内表达。TGF-β1可能通过Sp1信号转录途径调控Ⅰ型胶原的合成与降解,从而在近视巩膜重塑中发挥重要作用。但其具体机制还有待进一步研究。
[Abstract]:Aim: to study the expression of Sp1 as a downstream signal transcription gene of TGF- 尾 1 in myopic scleral remodeling. To investigate the relationship between the expression of Sp1 protein and m RNA and (form deprived myopia,FDM in experimental form deprivation myopia. To explore the role of TGF- 尾 1-Sp1-Collagen I signaling pathway in scleral collagen remodeling in myopia. Methods: 75 healthy tricolor guinea pigs about one week old were selected, and 6 translucent latex balloon was used as eye mask to cover the eyes of guinea pigs in order to induce the animal model of form deprivation myopia. 75 guinea pigs were randomly divided into normal control group (50 eyes), form deprivation group (FDM group, 50 eyes) and self control group (50 eyes). The self-control group was the exposed eye of guinea pigs in the experimental group. The method was to use translucent latex balloon to make a headset according to the shape of the head of guinea pig, covering the left eye of guinea pig, exposing its right eye, mouth and nose, and both ears. This method has many advantages, such as stable, harmless, no change of cornea pressure and temperature, etc. The diopter and axial length of the eyes were measured before (0 weeks), 2 weeks, 4 weeks, 6 weeks and 4 weeks to cover 1 week (4 / -1 week). Immunohistochemical technique and RT-PCR method were used to detect the dynamic expression of SP1 in the sclera of guinea pig orthopia and experimental myopia during the period of 0 weeks, 2 weeks, 4 weeks, 6 weeks and 4 weeks of induction, respectively. The relationship between proto synthesis and expression. Results: in the form deprivation group (FDM group), the hyperopia eyes (2.09 卤0.31D, -1.23 卤0.69) D4 weeks (-4.17 卤0.59) D, the hyperopia eyes (-4.30 卤0.58D) and the shaded 6 weeks (-7.07 卤0.56) D gradually became myopia after 4 weeks of covering (-4.30 卤0.58) D and 6 weeks of covering (-7.07 卤0.56) D. The axial length of the eye was prolonged [the length of the anterior eye axis: (5.93 卤0.39) mm;2 week: (6.62 卤0.36) mm;4 week: (7.30 卤0.34) mm;4 week after 1 week eye axis length: (7.41 卤0.36) mm;6 week: (7.99 卤0.32) mm,P0.05]. Immunohistochemical results showed that the positive expression of Sp1 in sclera of form-deprived (FDM) group was weaker than that of control group, and the positive expression of Sp1 decreased with the prolongation of shading time (p0.05). The results of RT-PCR showed that, compared with the control group, the positive expression of Sp1 in the sclera was significantly lower than that in the control group (p0.05). In form deprivation group, m RNA expression of Sp1 was down-regulated (p0.05), and the expression of, m RNA was gradually down-regulated (p0.05) over time. Conclusion: Sp1 is a downstream signal transcription gene of TGF- 尾 1 and expressed in the sclera of guinea pigs. TGF- 尾 1 may play an important role in myopia scleral remodeling by regulating the synthesis and degradation of type I collagen through Sp1 signal transcriptional pathway. But its specific mechanism still needs further research.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R778.11

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