【摘要】： 目的:通過建立大鼠糖尿病視網(wǎng)膜病變模型,將血管生成素-1(Ang-1)注射于大鼠眼玻璃體腔內(nèi),觀察Ang-1對(duì)糖尿病大鼠視網(wǎng)膜血管內(nèi)皮細(xì)胞生長(zhǎng)因子(VEGF)及新生血管形成的影響,為Ang-1治療糖尿病視網(wǎng)膜病變提供有實(shí)用價(jià)值的實(shí)驗(yàn)依據(jù)。 方法:48只健康SD大鼠,隨機(jī)抽取16只作為正常對(duì)照組(A組),標(biāo)準(zhǔn)條件飼養(yǎng)。另外32只大鼠以65mg/kg的劑量行左下腹腔注射10%鏈尿佐菌素溶液(STZ),并于24小時(shí)后檢測(cè)血糖,將餐后血糖濃度持續(xù)穩(wěn)定大于16.7mmol/L的大鼠定為糖尿病模型,相同條件喂養(yǎng)4個(gè)月后行眼底血管熒光造影檢查大鼠出現(xiàn)視網(wǎng)膜病變(DR)即為成模,當(dāng)即隨機(jī)分為兩組(每組16只),陽性對(duì)照組(B組)和Ang-1治療組(C組)。陽性對(duì)照組(B組)行雙眼玻璃體腔注射磷酸鹽緩沖生理鹽水(PBS)5μl,Ang-1治療組(C組)行雙眼玻璃體腔注射160μg/mL Ang-1 5μl。三天后重復(fù)上述處理一次,繼續(xù)觀察三天后處死三組大鼠取視網(wǎng)膜組織免疫組化法檢測(cè)比較VEGF的表達(dá),病理切片HE染色比較突出視網(wǎng)膜內(nèi)界膜內(nèi)皮細(xì)胞核數(shù)。 結(jié)果:(1)免疫組化法檢查:陽性對(duì)照組(B組)與正常對(duì)照組(A組)比較視網(wǎng)膜VEGF免疫組化染色表達(dá)水平明顯增強(qiáng),積分光密度測(cè)定值比較差異有統(tǒng)計(jì)學(xué)意義(P0.05);Ang-1治療組(C組)與正常對(duì)照組(A組)比較視網(wǎng)膜VEGF免疫組化染色表達(dá)水平明顯增強(qiáng),積分光密度測(cè)定值比較差異有統(tǒng)計(jì)學(xué)意義(P0.05);Ang-1治療組(C組)與陽性對(duì)照組(B組)相比VEGF免疫組化染色表達(dá)較弱,積分光密度值比較差異有統(tǒng)計(jì)學(xué)意義(P0.05);(2)HE染色病理學(xué)切片示:陽性對(duì)照組(B組)與正常對(duì)照組(A組)比較突破視網(wǎng)膜內(nèi)界膜內(nèi)皮細(xì)胞核數(shù)明顯增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05);Ang-1治療組(C組)與正常對(duì)照組(A組)比較突破視網(wǎng)膜內(nèi)界膜內(nèi)皮細(xì)胞核數(shù)明顯增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05);Ang-1治療組(C組)與陽性對(duì)照組(B組)比較突破視網(wǎng)膜內(nèi)界膜內(nèi)皮細(xì)胞核數(shù)減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論:大鼠出現(xiàn)糖尿病視網(wǎng)膜病變時(shí)視網(wǎng)膜會(huì)產(chǎn)生大量的VEGF,將Ang-1注射于玻璃體腔內(nèi)對(duì)STZ誘導(dǎo)的糖尿病視網(wǎng)膜病變大鼠VEGF的表達(dá)有一定的抑制作用,并有效的降低了視網(wǎng)膜新生血管的生成,從而對(duì)糖尿病視網(wǎng)膜病變起到一定的治療作用。
[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.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R774.1

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