【摘要】 目的:研究抗成纤维细胞生长因子(FGF-2)纳米抗体对碱烧伤诱导的大鼠角膜血管生成的治疗作用。方法:将SD大鼠分为:假手术组(Sham),模型组(Model,直径为3 mm的浸有1 mol/L NaOH溶液圆形滤纸贴于大鼠眼角膜中央处30 s,制备大鼠碱烧伤血管生成模型)和治疗组(Treatment,术后7天至21天用3 mg/mL的抗FGF-2纳米抗体溶液滴眼,每日3次,每次10μL,共14天)。通过体视显微镜和CD31免疫组织化学染色计算大鼠角膜血管生成情况。实时荧光定量PCR、酶联免疫吸附测定和免疫组织化学染色3种方法检测抗血管内皮生长因子(VEGF)和FGF-2的mRNA和蛋白表达水平。结果:(1)血管:治疗组较模型组的面积显著减少,血管管腔较窄(P<0.05),在药物干预14天后,差异最为显著;(2)FGF-2的mRNA和蛋白表达水平:模型组与治疗组的结果相近(P>0.05);(3)VEGF的mRNA和蛋白表达水平:治疗组显著高于模型组(P<0.05)。此外,假手术组的持续给药也使得VEGF表达显著增加(P<0.05)。结论:抗FGF-2纳米抗体可抑制由碱烧伤诱导的角膜血管新生,但也使得正常大鼠角膜或病理大鼠角膜的VEGF表达水平代偿性升高。更多还原

【Abstract】 AIM: To investigate the possible use of anti-FGF-2 nanobody for the treatment of pathological neovascularization. METHODS: SD rats were divided into a sham operation group, a control group(3 mm diameter circular filter paper soaked with 1 mol/L NaOH solution was applied to the central part of the cornea of rats for 30 s to prepare the rat model of alkali-burn angiogenesis) and a treatment group(treated with a drop of 3 mg/mL anti-FGF-2 nanobody 7 days after the operation. Repeat application 3 x/day for 14 days). Corneal angiogenesis was measured by stereoscopic microscopy and CD31 immunohistochemical staining. The mRNA and protein expression levels of VEGF and FGF-2 were detected by quantitative fluorescence PCR(qPCR), enzyme-linked immunosorbent assay(ELISA) and immunohistochemistry.RESULTS:(1) Blood vessel: The area of the treatment group was significantly reduced compared with the model group, and the vascular lumen was narrower(P<0.05). The difference was the most significant after 14 days of drug intervention;(2) Expression level of FGF-2 mRNA and protein: the model group had similar results to the treatment group(P>0.05);(3) Expression levels of VEGF mRNA and protein: The treatment group was significantly higher than the model group(P<0.05). In addition, the expression of VEGF also increased significantly in the continuous administration of the sham operation group. CONCLUSION: Anti-FGF-2 nanobody can be used for the treatment of angiogenesis. However, the expressions of VEGF will compensatorily increase after blocking FGF-2 in normal or pathological rats.更多还原