实验性脉络膜新生血管COX-2、CD31的表达变化及塞来昔布对其影响的研究

发布时间：2018-02-23 18:45

本文关键词： 脉络膜新生血管环氧化酶-2 血小板内皮细胞黏附分子　出处：《昆明医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：观察环氧合酶-2(cyclooxygenase-2,CoX-2)和血小板内皮细胞黏附分子(CD31),在激光诱导的C57BL/6小鼠脉络膜新生血管(choroidal noevasculariza-tion, CNV)生成中表达的时相的变化,以及COX-2抑制剂塞来昔布对脉络膜新生血管的作用机制,进一步探讨CNV中的发病机制和治疗方法。方法：45只C57BL/6小鼠,随机分成正常对照组、激光光凝组、塞来昔布治疗组,激光光凝组采用多波长氪离子激光过量光凝诱导脉络膜新生血管。塞来昔布组(激光光凝前一周开始用药,经口灌胃,剂量为100Mg/kg,1次/d,直到实验结束)。正常对照组于光凝前取标本,光凝组及塞来昔布治疗组于光凝后7天、14天、21天、28天取标本。取材前腹腔注射戊巴比妥注射液lml/kg麻醉,处死、迅速摘除眼球、取材、石蜡包埋,、HE染色、COX-2和CD31免疫组化染色。光镜观察光凝损伤处视网膜脉络膜组织的变化,采用Image-ProPlus6.0软件对COX-2阳性染色的平均光密度(mean optical density,MOD)进行测量,所有数据均采用SPSS12.0for Windows软件包进行处理,进行统计学分析。结果：1、激光光凝组中,21天组微血管密度(MVD)高于7天组、14天组,其差异具有统计学意义(P0.05),MVD在14天组和28天组之间无明显差异(P0.05)。2、正常对照组分别与光凝组、塞来昔布治疗组不同时间点的比较,治疗组7天波动于正常对照组左右,差异无统计学意义,P0.05；其他的均高于正常对照组,P0.05,其差异有统计学意义。3、光凝组和塞来昔布治疗组组内各时间点之间比较,差异均有统计学意义(P0.05)。4、光凝后7天COX-2的表达略增强,14天时表达明显增强,21天呈强阳性表达,28天时呈继续减弱趋势,塞来昔布组与光凝组同一时间点比较,7天两组无明显差异,无统计学意义(P0.05),在14天,21天,28天COX-2阳性表达的量较单纯光凝组降低,其差异有统计学意义(P0.05)。结论：1、该实验利用氪激光成功诱导了C57BL/6小鼠脉络膜新生血管模型。2、在激光诱导的脉络膜新生血管形成中,局部视网膜脉络膜组织中COX-2蛋白的表达是增高的3、塞来昔布可以降低激光诱导的脉络膜新生血管中COX-2的表达,从而在一定程度上减少CNV的形成。
[Abstract]:Aim: to observe the changes of expression of cyclooxygenase-2 CoX-2) and platelet endothelial cell adhesion molecule (CD31) in laser-induced choroidal noevasculariza-tion (CNV) production in C57BL6 mice. The mechanism of COX-2 inhibitor celecoxib on choroidal neovascularization and the pathogenesis and treatment of CNV were discussed. Methods 45 C57BL / 6 mice were randomly divided into normal control group, laser photocoagulation group and celecoxib treatment group. In laser photocoagulation group, choroidal neovascularization was induced by excessive photocoagulation with multi-wavelength krypton laser. The dose was 100 mg / kg / kg / d until the end of the experiment. The normal control group took the specimen before photocoagulation, the photocoagulation group and the celecoxib treatment group took the specimen at 7 days and 14 days after photocoagulation and 21 days and 28 days after photocoagulation. The sample was injected intraperitoneally with pentobarbital injection lml/kg anaesthesia before sampling and was executed. The eyeball was removed quickly, and the paraffin-embedded HE staining was used to detect COX-2 and CD31 immunohistochemical staining. The changes of retinal choroidal tissue were observed under light microscope, and the mean optical density (Mod) of COX-2 positive staining was measured by Image-ProPlus6.0 software. All the data were processed by SPSS12.0for Windows software package and analyzed statistically. Results the MVDs in the laser photocoagulation group were significantly higher than those in the 7th day group. There was no significant difference in MVD between the 14 day group and the 28 day group. The normal control group and the light coagulation group had no significant difference in MVD, and the normal control group and the photocoagulation group had no significant difference in MVD between the 14 day group and the 28 day group, the normal control group and the photocoagulation group, respectively. The comparison of different time points in celecoxib treatment group showed that the treatment group fluctuated about 7 days compared with the normal control group. The difference was not statistically significant (P 0.05), the other was higher than that of the normal control group (P 0.05), the difference was statistically significant. The difference was significant between the photocoagulation group and celecoxib treatment group at different time points. The expression of COX-2 was slightly enhanced on the 7th day after photocoagulation. The expression of COX-2 was significantly increased on the 14th day and continued to weaken on the 21st day after photocoagulation. There was no significant difference between the two groups at the same time point in the celecoxib group and the photocoagulation group at the same time point, and there was no significant difference between the two groups at the same time point. There was no significant difference in the expression of COX-2 between the two groups on the 14th day and 21st day, and the positive expression of COX-2 was significantly lower than that of the photocoagulation group, and the difference was statistically significant (P 0.05). Conclusion: in this experiment, the choroidal neovascularization model of C57BL / 6 mice was successfully induced by Krypton laser in the laser-induced choroidal neovascularization. The expression of COX-2 protein was increased by 3% in the local retinal choroid tissue. Celecoxib could reduce the expression of COX-2 in the laser-induced choroidal neovascularization and thus reduce the formation of CNV to a certain extent.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R773.4

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