【作者】 张欣；

【导师】 周芳；

【作者基本信息】 山东大学 ， 眼科学， 2006， 硕士

【摘要】 目的：通过观察生长抑素类似物奥曲肽在实验性脉络膜新生血管(CNV)动物模型中对碱性成纤维细胞生长因子(b-FGF)和转化生长因子(TGF-β)的调控作用，研究奥曲肽对脉络膜新生血管的抑制作用。 方法：激光建立C57BL／6小鼠CNV模型，建模后7d、14d、21d和28d为观察时间点。135只C57BL／6小鼠中3只为正常对照，132只用于实验，每个时间点各3只作为疾病对照，余120只作为治疗组分为P0组和P7组：激光制作CNV模型当天开始用药为P0组，光凝后7天开始用药为P7组。P0组和P7组又各分为3组：单纯皮下注射组、单纯球后注射组和皮下+球后联合注射组，每组每个时间点各5只小鼠。皮下注射和球后注射奥曲肽的剂量均为50μg／kg体重，1次／d，连用14天。分别于激光照射后7d、14d、21d和28d处死小鼠，摘除眼球，行HE染色，采用逆转录多聚酶链反应(RT-PCR)检测不同时间点不同注药方式视网膜脉络膜中b-FGF和TGF-βmRNA的变化，分析表达强度，计算相对系数：相对系数(％)=标志物表达强度／β-actin表达强度。数据经SPSS12.0统计学软件处理，采用两样本t检验。P＜0.05，差异有显著性；P＜0.01，差异有极显著性。 结果：实验用鼠132只，造模成功117只。造影结果显示，疾病对照组照射7d、2w和3w时，可见大部分光凝斑处有荧光渗漏，呈片状，随时间延长，荧光范围增大，亮度增强，证实有CNV形成。HE染色显示，疾病对照组激光光凝术后7d，光凝斑处色素紊乱、缺失，脉络膜增厚，Bruch膜中断，视网膜色素上皮层下可见新生血管，提示CNV形成。 RT-PCR检查结果显示，视网膜和脉络膜组织中均有b-FGF和TGF-βmRNA的表达。 P0组单纯皮下用药早期不能抑制b-FGF增加，晚期b-FGF的阳性表达明显减少。球后注射和联合用药早期能抑制CNV膜中b-FGF的阳性表达。P7组皮下更多还原

【Abstract】 Objective: To observe the effect of somatostatin analogue octreotide on b-FGF,TGF-pin the experimental choroidal neovascularization (CNV) of mice, investigating octreotide as an inhibitor on the formation of CNV. Methods: Diode laser was applied to the eyes in C57BL/6 mice to induce choroidal neovascularization(CNV). We chose post-photocoagulation 7d, 14d, 21d and 28d as observed time point. In all 135 mice, 3 mice as normal control, 3 in one observed point as diseased control, 120 mice treated with octreotide were divided into 2 groups: group P0 was treated as soon as finished laser photocoagulation, group P7 was treated 7 days after photocoagulation. Each groups had three sub-groups: simply treated with subcutaneous injection of octreotide, with retrobulbar injection, or two methods combined. The dose of octreotide was 50μg/kg, qd×l4d. At each observed point, mice were sacrificed, the eyes were removed, Reverse transcription poly chain reaction (RT-PCR) was used to detect the changes of b-FGF or TGF-β mRNA by different medications at different observed point, analysising the expression intensity and relative coefficient.All data were analyzed with SPSS 12.0, using t analysis. P<0.05, difference was significant; P<0.01, difference was extremely significant.Results: Fluorescein leakage and histopathological evidence of CNV were found one week after diode laser photocoagulation in C57BL/6J mice.RT-PCR showed that b-FGF or TGF-β mRNA were expressed in the sclera-choroid-retina complex.RT-PCR showed that the positive expression of b-FGF in CNVMs could not be inhibited by subcutaneous injection of octreotide in the early stage in group P0. In the late stage, it significantly decreased. In this group, retrobulbar injection or two methods combined were both effective. In group P7, subcutaneous injection of octreotide could not inhibit the expression of b-FGF in CNVMs, the other methods更多还原