本文選題：骨形態(tài)發(fā)生蛋白 + 鞏膜��； 參考：《青島大學(xué)》2011年博士論文

【摘要】：目的：檢測骨形態(tài)發(fā)生蛋白(BMPs)在人、大鼠以及豚鼠鞏膜中的表達(dá)；建立豚鼠形覺剝奪近視眼(FDM)動物模型,觀察BMPs在豚鼠后鞏膜中的表達(dá)變化；原代培養(yǎng)人鞏膜成纖維細(xì)胞(HSF),觀察BMP-2對鞏膜成纖維細(xì)胞的作用；臨床觀察高度近視患者植入多焦點(diǎn)人工晶體的術(shù)后視覺質(zhì)量。方法：應(yīng)用RT-PCR和免疫熒光的方法分別檢測人、大鼠和豚鼠后鞏膜中BMPs的表達(dá)；連續(xù)形覺剝奪14天后建立豚鼠形覺剝奪近視動物模型,實驗前后分別測量屈光度和眼軸。RT-PCR和western-blot檢測豚鼠實驗性近視眼后鞏膜中BMPs的表達(dá)變化；原代培養(yǎng)HSF,觀察各種濃度BMP-2對HSF的作用：用四甲基偶氮唑藍(lán)(MTT)法檢測細(xì)胞增殖能力,流式細(xì)胞儀檢測細(xì)胞的周期,RT-PCR和western-blot檢測膠原蛋白Ⅰ和蛋白聚糖以及激活的信號轉(zhuǎn)導(dǎo)通路蛋白的變化,細(xì)胞表型以及激活的信號轉(zhuǎn)導(dǎo)蛋白進(jìn)一步由抗α-SMA和抗phospho-smadl/5/8的免疫熒光法確定。36例高度近視白內(nèi)障患者行超聲乳化聯(lián)合人工晶體植入,按照植入人工晶體的類型分為3組,AcrySof IQ ReSTOR SN6AD3(+4D組),AcrySof IQ ReSTOR SN6AD1(+3D組)和AcrySof IQ SN60WF(單焦點(diǎn)組),每組12例,24眼。測量術(shù)后的遠(yuǎn)、中、近視力,遠(yuǎn)視力調(diào)制傳遞函數(shù)(MTF),焦點(diǎn)深度,并問卷調(diào)查術(shù)后滿意度等。結(jié)果：(1)人后鞏膜表達(dá)BMP-2、BMP-4、BMP-5和BMP-7,大鼠只表達(dá)BMP-7,而豚鼠表達(dá)BMP-2,BMP-4和BMP-5。(2)實驗后FDM組實驗眼的屈光度和眼軸分別為-0.48±0.51D和8.30±0.05mm(p0.001分別相對于正常對照眼和組內(nèi)對側(cè)對照眼)。(3)實驗眼后鞏膜中BMP-2和BMP-5的表達(dá)在基因和蛋白水平都相對于對側(cè)對照眼明顯降低,而BMP-4表達(dá)沒有明顯改變。(4)100ng/ml BMP-2明顯促進(jìn)HSF的增殖,經(jīng)BMP-2誘導(dǎo)后,處于S期的HSF比率明顯升高,而G1期明顯下降(p0.05),免疫熒光發(fā)現(xiàn)更多的細(xì)胞轉(zhuǎn)化為肌成纖維細(xì)胞；培養(yǎng)72小時后,膠原蛋白I(Collagen I)和蛋白聚糖(Aggrecan)的表達(dá)明顯升高(p0.05),信號轉(zhuǎn)導(dǎo)蛋白phospho-smadl/5/8的表達(dá)在BMP-2刺激1小時后明顯升高。(5)術(shù)后6個月,兩多焦點(diǎn)人工晶體組的術(shù)后中、近距離視力明顯高于單焦點(diǎn)組(p0.01),+3D組的中間距離視力高于+4D組(p0.05),而+4D組的近距離視力高于+3D組(p0.05)：5mm瞳孔下,三組的MTF沒有明顯差別；兩組多焦點(diǎn)人工晶體植入組的術(shù)后滿意度和脫鏡率一樣高。結(jié)論：(1)人和豚鼠鞏膜都表達(dá)BMP-2、BMP-4和BMP-5。(2)BMP-2和BMP-5在實驗性近視豚鼠后鞏膜中的表達(dá)降低。(3)在體外BMP-2能夠促進(jìn)HSF增殖和分化,并促進(jìn)細(xì)胞外基質(zhì)膠原蛋白Ⅰ和蛋白聚糖的合成。(4)BMP-2作用于HSF可能是通過Smad信號轉(zhuǎn)導(dǎo)通路。(5)高度近視白內(nèi)障患者行超聲乳化聯(lián)合多焦點(diǎn)人工晶體植入可以給患者提供良好的遠(yuǎn)、中、近功能性視力,達(dá)到術(shù)后較高的滿意度和脫鏡率。
[Abstract]:Objective: to detect the expression of bone morphogenetic protein (BMPs) in the sclera of human, rat and guinea pig, to establish the animal model of form-deprivation myopia and to observe the change of BMPs expression in the posterior sclera of guinea pig. To observe the effect of BMP-2 on scleral fibroblasts in primary culture of human scleral fibroblasts, and to observe the postoperative visual quality of high myopia patients implanted with multi-focus intraocular lens. Methods: the expression of BMPs in the posterior sclera of human, rat and guinea pig was detected by RT-PCR and immunofluorescence, and the myopia model was established after 14 days of continuous form deprivation. The changes of BMPs expression in posterior sclera of experimental myopia in guinea pigs were measured by diopter, eye axis. RT-PCR and western-blot before and after the experiment, the effects of BMP-2 at various concentrations on HSF were observed by primary culture: the proliferation of cells was detected by tetramethylazolazolam (MTT) method, and the expression of BMPs in the sclera of guinea pig experimental myopia was measured by RT-PCR and western-blot before and after the experiment. The changes of collagen 鈪，

本文編號：2016298