骨形态发生蛋白在实验性近视后巩膜重塑中的作用以及高度近视相关临床治疗观察

发布时间：2018-06-14 00:53

  本文选题：骨形态发生蛋白 + 巩膜　； 参考：《青岛大学》2011年博士论文

【摘要】：目的：检测骨形态发生蛋白(BMPs)在人、大鼠以及豚鼠巩膜中的表达；建立豚鼠形觉剥夺近视眼(FDM)动物模型,观察BMPs在豚鼠后巩膜中的表达变化；原代培养人巩膜成纤维细胞(HSF),观察BMP-2对巩膜成纤维细胞的作用；临床观察高度近视患者植入多焦点人工晶体的术后视觉质量。方法：应用RT-PCR和免疫荧光的方法分别检测人、大鼠和豚鼠后巩膜中BMPs的表达；连续形觉剥夺14天后建立豚鼠形觉剥夺近视动物模型,实验前后分别测量屈光度和眼轴。RT-PCR和western-blot检测豚鼠实验性近视眼后巩膜中BMPs的表达变化；原代培养HSF,观察各种浓度BMP-2对HSF的作用：用四甲基偶氮唑蓝(MTT)法检测细胞增殖能力,流式细胞仪检测细胞的周期,RT-PCR和western-blot检测胶原蛋白Ⅰ和蛋白聚糖以及激活的信号转导通路蛋白的变化,细胞表型以及激活的信号转导蛋白进一步由抗α-SMA和抗phospho-smadl/5/8的免疫荧光法确定。36例高度近视白内障患者行超声乳化联合人工晶体植入,按照植入人工晶体的类型分为3组,AcrySof IQ ReSTOR SN6AD3(+4D组),AcrySof IQ ReSTOR SN6AD1(+3D组)和AcrySof IQ SN60WF(单焦点组),每组12例,24眼。测量术后的远、中、近视力,远视力调制传递函数(MTF),焦点深度,并问卷调查术后满意度等。结果：(1)人后巩膜表达BMP-2、BMP-4、BMP-5和BMP-7,大鼠只表达BMP-7,而豚鼠表达BMP-2,BMP-4和BMP-5。(2)实验后FDM组实验眼的屈光度和眼轴分别为-0.48±0.51D和8.30±0.05mm(p0.001分别相对于正常对照眼和组内对侧对照眼)。(3)实验眼后巩膜中BMP-2和BMP-5的表达在基因和蛋白水平都相对于对侧对照眼明显降低,而BMP-4表达没有明显改变。(4)100ng/ml BMP-2明显促进HSF的增殖,经BMP-2诱导后,处于S期的HSF比率明显升高,而G1期明显下降(p0.05),免疫荧光发现更多的细胞转化为肌成纤维细胞；培养72小时后,胶原蛋白I(Collagen I)和蛋白聚糖(Aggrecan)的表达明显升高(p0.05),信号转导蛋白phospho-smadl/5/8的表达在BMP-2刺激1小时后明显升高。(5)术后6个月,两多焦点人工晶体组的术后中、近距离视力明显高于单焦点组(p0.01),+3D组的中间距离视力高于+4D组(p0.05),而+4D组的近距离视力高于+3D组(p0.05)：5mm瞳孔下,三组的MTF没有明显差别；两组多焦点人工晶体植入组的术后满意度和脱镜率一样高。结论：(1)人和豚鼠巩膜都表达BMP-2、BMP-4和BMP-5。(2)BMP-2和BMP-5在实验性近视豚鼠后巩膜中的表达降低。(3)在体外BMP-2能够促进HSF增殖和分化,并促进细胞外基质胶原蛋白Ⅰ和蛋白聚糖的合成。(4)BMP-2作用于HSF可能是通过Smad信号转导通路。(5)高度近视白内障患者行超声乳化联合多焦点人工晶体植入可以给患者提供良好的远、中、近功能性视力,达到术后较高的满意度和脱镜率。
[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，

本文编号：2016298