【摘要】：目的:研究近視鞏膜重塑中Sp1作為TGF-β1下游信號轉(zhuǎn)錄基因的表達(dá)情況。探討Sp1蛋白分子及m RNA的表達(dá)與實(shí)驗(yàn)性形覺剝奪性近視(form deprived myopia,FDM)之間的關(guān)系。以期揭示TGF-β1-Sp1-CollagenⅠ信號途徑在近視鞏膜膠原重塑中的作用。方法:選取出生1周左右的健康三色豚鼠75只,利用6號半透明乳膠氣球作為眼罩遮蓋豚鼠的眼睛以誘導(dǎo)形覺剝奪性近視動物模型。將75只豚鼠隨機(jī)分為正常對照組(50眼)、形覺剝奪(FDM組,50眼)組和自身對照組(50眼),正常對照組是未做任何處理的豚鼠,FDM組是實(shí)驗(yàn)組的豚鼠遮蓋眼,自身對照組是實(shí)驗(yàn)組豚鼠的暴露眼,方法是使用半透明的乳膠氣球,依據(jù)豚鼠的頭部形狀制作成頭套,遮蓋豚鼠的左眼,暴露其右眼、口鼻及雙耳。這種方法具有穩(wěn)定、無害、不會對角膜產(chǎn)生壓力和溫度變化等諸多的優(yōu)勢。分別測量記錄實(shí)驗(yàn)豚鼠在未遮蓋前(0周)、遮蓋2周、4周、6周及遮蓋4周去遮蓋1周(4/-1周)時(shí)的雙眼屈光度及眼軸長度。采用免疫組織化學(xué)技術(shù)、RT-PCR法檢測豚鼠正視眼和實(shí)驗(yàn)性近視眼鞏膜中分別在遮蓋0周、2周、4周、6周及誘導(dǎo)4周去誘導(dǎo)1周時(shí)間段內(nèi)SP1的動態(tài)表達(dá)變化以及與Ⅰ型膠原合成與表達(dá)的變化關(guān)系。結(jié)果:形覺剝奪組(FDM組)由遮蓋前(0周)的遠(yuǎn)視眼(+2.09±0.31)D到遮蓋2周(-1.23±0.69)D、4周(-4.17±0.59)D、遮蓋4周去遮蓋1周(-4.30±0.58)D及遮蓋6周(-7.07±0.56)D后逐漸變成近視眼,且隨著時(shí)間延長,近視度數(shù)不斷加深,眼軸也逐漸延長[遮蓋前眼軸長:(5.93±0.39)mm;2周時(shí)眼軸長:(6.62±0.36)mm;4周時(shí)眼軸長:(7.30±0.34)mm;4周去1周眼軸長:(7.41±0.36)mm;6周時(shí)眼軸長:(7.99±0.32)mm,P0.05]。免疫組化結(jié)果顯示,形覺剝奪(FDM)組鞏膜有Sp1陽性表達(dá),其表達(dá)均弱于兩組對照組,且隨著遮蓋時(shí)間的延長,Sp1的陽性表達(dá)在不斷減弱(p0.05)。RT-PCR結(jié)果認(rèn)為,和兩組對照組相比,形覺剝奪組Sp1的m RNA表達(dá)均下調(diào)(p0.05),且隨著時(shí)間的延長,m RNA的表達(dá)也在逐漸下調(diào)(p0.05)。結(jié)論:Sp1是TGF-β1的下游信號轉(zhuǎn)錄基因,且在豚鼠鞏膜內(nèi)表達(dá)。TGF-β1可能通過Sp1信號轉(zhuǎn)錄途徑調(diào)控Ⅰ型膠原的合成與降解,從而在近視鞏膜重塑中發(fā)揮重要作用。但其具體機(jī)制還有待進(jìn)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 鹵0.31D, -1.23 鹵0.69) D4 weeks (-4.17 鹵0.59) D, the hyperopia eyes (-4.30 鹵0.58D) and the shaded 6 weeks (-7.07 鹵0.56) D gradually became myopia after 4 weeks of covering (-4.30 鹵0.58) D and 6 weeks of covering (-7.07 鹵0.56) D. The axial length of the eye was prolonged [the length of the anterior eye axis: (5.93 鹵0.39) mm;2 week: (6.62 鹵0.36) mm;4 week: (7.30 鹵0.34) mm;4 week after 1 week eye axis length: (7.41 鹵0.36) mm;6 week: (7.99 鹵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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R778.11

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