ANG-1对大鼠糖尿病视网膜病变新生血管形成的影响
发布时间：2018-08-30 08:26
【摘要】： 目的:通过建立大鼠糖尿病视网膜病变模型,将血管生成素-1(Ang-1)注射于大鼠眼玻璃体腔内,观察Ang-1对糖尿病大鼠视网膜血管内皮细胞生长因子(VEGF)及新生血管形成的影响,为Ang-1治疗糖尿病视网膜病变提供有实用价值的实验依据。 方法:48只健康SD大鼠,随机抽取16只作为正常对照组(A组),标准条件饲养。另外32只大鼠以65mg/kg的剂量行左下腹腔注射10%链尿佐菌素溶液(STZ),并于24小时后检测血糖,将餐后血糖浓度持续稳定大于16.7mmol/L的大鼠定为糖尿病模型,相同条件喂养4个月后行眼底血管荧光造影检查大鼠出现视网膜病变(DR)即为成模,当即随机分为两组(每组16只),阳性对照组(B组)和Ang-1治疗组(C组)。阳性对照组(B组)行双眼玻璃体腔注射磷酸盐缓冲生理盐水(PBS)5μl,Ang-1治疗组(C组)行双眼玻璃体腔注射160μg/mL Ang-1 5μl。三天后重复上述处理一次,继续观察三天后处死三组大鼠取视网膜组织免疫组化法检测比较VEGF的表达,病理切片HE染色比较突出视网膜内界膜内皮细胞核数。 结果:(1)免疫组化法检查:阳性对照组(B组)与正常对照组(A组)比较视网膜VEGF免疫组化染色表达水平明显增强,积分光密度测定值比较差异有统计学意义(P0.05);Ang-1治疗组(C组)与正常对照组(A组)比较视网膜VEGF免疫组化染色表达水平明显增强,积分光密度测定值比较差异有统计学意义(P0.05);Ang-1治疗组(C组)与阳性对照组(B组)相比VEGF免疫组化染色表达较弱,积分光密度值比较差异有统计学意义(P0.05);(2)HE染色病理学切片示:阳性对照组(B组)与正常对照组(A组)比较突破视网膜内界膜内皮细胞核数明显增加,差异有统计学意义(P0.05);Ang-1治疗组(C组)与正常对照组(A组)比较突破视网膜内界膜内皮细胞核数明显增加,差异有统计学意义(P0.05);Ang-1治疗组(C组)与阳性对照组(B组)比较突破视网膜内界膜内皮细胞核数减少,差异有统计学意义(P0.05)。 结论:大鼠出现糖尿病视网膜病变时视网膜会产生大量的VEGF,将Ang-1注射于玻璃体腔内对STZ诱导的糖尿病视网膜病变大鼠VEGF的表达有一定的抑制作用,并有效的降低了视网膜新生血管的生成,从而对糖尿病视网膜病变起到一定的治疗作用。
[Abstract]:Objective: to investigate the effect of Ang-1 on retinal vascular endothelial growth factor (VEGF) and angiogenesis in diabetic rats by injecting angiopoietin 1 (Ang-1) into vitreous cavity of rats with diabetic retinopathy. To provide a practical experimental basis for the treatment of diabetic retinopathy with Ang-1. Methods Sixteen healthy SD rats were randomly selected as normal control group (group A). The other 32 rats were injected with 10% streptozotocin solution (STZ),) intraperitoneally at the dose of 65mg/kg and blood glucose was detected 24 hours later. The rats whose postprandial blood glucose concentration was more stable than 16.7mmol/L were selected as diabetic model. After 4 months of feeding under the same conditions, retinopathy (DR) was established in rats by fundus fluorescein angiography. The rats were randomly divided into two groups (16 rats in each group), positive control group (B group) and Ang-1 treatment group (C group). The positive control group (group B) was injected with phosphate buffer saline (PBS) 5 渭 l Ang-1) into the vitreous cavity of both eyes (group C), and the group C received the injection of 160 渭 g/mL Ang-1 5 渭 l into the vitreous cavity. The above treatment was repeated three days later. After 3 days of observation, the three groups of rats were killed to detect the expression of VEGF by immunohistochemical method, and the number of endothelial nuclei in the inner boundary membrane of the retina was compared by HE staining in pathological sections. Results: (1) Immunohistochemistry: the expression of VEGF in the retina of the positive control group (group B) was significantly higher than that of the normal control group (group A). There was significant difference in integrated optical density (P0.05). The expression of VEGF immunohistochemical staining in retina of group C (group C) was significantly higher than that of group A (normal control group), and the expression of VEGF in retina was significantly higher than that in group C (P 0.05). Compared with the positive control group (group B), the expression of VEGF immunohistochemical staining was weaker in group C than in group C (P0.05). The difference of integrated optical density was statistically significant (P0.05); (2) HE staining pathological sections showed that the number of endothelial nuclei breaking through the inner boundary of retina increased significantly in the positive control group (group B) and the normal control group (group A). The difference was statistically significant (P0.05). Compared with the control group (group A), the number of the endothelial nucleus of the inner boundary of the retina was significantly increased in the Ang-1 treatment group (C group). The difference was statistically significant (P0.05). Compared with the positive control group (group B), the number of endothelium nucleus of retinal membrane breakthrough in Ang-1 treatment group (P 0.05) was significantly lower than that in control group (P 0.05). Conclusion: when diabetic retinopathy occurs in rats, a large amount of VEGF, will be produced in the retina. Injection of Ang-1 into the vitreous cavity can inhibit the expression of VEGF in STZ induced diabetic retinopathy rats. And effectively reduce the formation of retinal neovascularization, which plays a role in the treatment of diabetic retinopathy.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R774.1
本文编号：2212531
[Abstract]:Objective: to investigate the effect of Ang-1 on retinal vascular endothelial growth factor (VEGF) and angiogenesis in diabetic rats by injecting angiopoietin 1 (Ang-1) into vitreous cavity of rats with diabetic retinopathy. To provide a practical experimental basis for the treatment of diabetic retinopathy with Ang-1. Methods Sixteen healthy SD rats were randomly selected as normal control group (group A). The other 32 rats were injected with 10% streptozotocin solution (STZ),) intraperitoneally at the dose of 65mg/kg and blood glucose was detected 24 hours later. The rats whose postprandial blood glucose concentration was more stable than 16.7mmol/L were selected as diabetic model. After 4 months of feeding under the same conditions, retinopathy (DR) was established in rats by fundus fluorescein angiography. The rats were randomly divided into two groups (16 rats in each group), positive control group (B group) and Ang-1 treatment group (C group). The positive control group (group B) was injected with phosphate buffer saline (PBS) 5 渭 l Ang-1) into the vitreous cavity of both eyes (group C), and the group C received the injection of 160 渭 g/mL Ang-1 5 渭 l into the vitreous cavity. The above treatment was repeated three days later. After 3 days of observation, the three groups of rats were killed to detect the expression of VEGF by immunohistochemical method, and the number of endothelial nuclei in the inner boundary membrane of the retina was compared by HE staining in pathological sections. Results: (1) Immunohistochemistry: the expression of VEGF in the retina of the positive control group (group B) was significantly higher than that of the normal control group (group A). There was significant difference in integrated optical density (P0.05). The expression of VEGF immunohistochemical staining in retina of group C (group C) was significantly higher than that of group A (normal control group), and the expression of VEGF in retina was significantly higher than that in group C (P 0.05). Compared with the positive control group (group B), the expression of VEGF immunohistochemical staining was weaker in group C than in group C (P0.05). The difference of integrated optical density was statistically significant (P0.05); (2) HE staining pathological sections showed that the number of endothelial nuclei breaking through the inner boundary of retina increased significantly in the positive control group (group B) and the normal control group (group A). The difference was statistically significant (P0.05). Compared with the control group (group A), the number of the endothelial nucleus of the inner boundary of the retina was significantly increased in the Ang-1 treatment group (C group). The difference was statistically significant (P0.05). Compared with the positive control group (group B), the number of endothelium nucleus of retinal membrane breakthrough in Ang-1 treatment group (P 0.05) was significantly lower than that in control group (P 0.05). Conclusion: when diabetic retinopathy occurs in rats, a large amount of VEGF, will be produced in the retina. Injection of Ang-1 into the vitreous cavity can inhibit the expression of VEGF in STZ induced diabetic retinopathy rats. And effectively reduce the formation of retinal neovascularization, which plays a role in the treatment of diabetic retinopathy.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R774.1
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