重組人內皮抑素對氧誘導視網膜新生血管抑制作用的研究
發(fā)布時間:2018-06-04 13:27
本文選題:視網膜新生血管 + 內皮抑素; 參考:《石河子大學》2013年碩士論文
【摘要】:目的:探討重組人血管內皮抑素(恩度,ENDOSTAR)對高氧誘導的小鼠視網膜病變(oxygen-inducedretinopathy,OIR)模型中新生血管的抑制作用 方法:將60只C57BL/6J小鼠隨機分為正常對照組(A組),高氧對照組(B組),生理鹽水對照組(C組),恩度干預組(D組),每組15只,除正常對照組(A組)外,均建立OIR模型。正常對照組(A組),高氧對照組(B組)小鼠不做處理,生理鹽水對照組(C組)于第12d給予腹腔注射0.1mlNS,恩度干預組(D組)于第12d給予腹腔注射rh.ES注射液0.1ml,連續(xù)注射5d,鼠齡17d時,視網膜組織切片行HE染色計數突破內界膜的新生血管內皮細胞核數目;應用免疫組織化學染色法檢測視網膜中VEGF的表達。 結果:HE染色正常對照組(A組)平均每張切片切片突破內界膜的內皮細胞細胞核個數為(0.97±0.04)個,高氧對照組(B組)平均每張切片突破內界膜的內皮細胞細胞核個數為(24.30±1.73)個。生理鹽水對照組(C組)平均每張切片突破內界膜的內皮細胞細胞核個數為個數為(22.58±1.54)個。恩度干預組(D組)突破視網膜內界膜的血管內皮細胞核數目為(3.38±0.27)個。高氧對照組(B組)和生理鹽水對照組(C組)突破內界膜的新生血管內皮細胞核數目明顯多于正常對照組(A組),差異均有顯著統(tǒng)計學意義(P<0.01);恩度干預組(D組)視網膜突破內界膜的新生血管內皮細胞核數目明顯減少,與高氧對照組(B組)和生理鹽水對照組(C組)比較,差異均有顯著統(tǒng)計學意義(P<0.01);恩度干預組(D組)與正常對照組(A組)比較差異具有顯著統(tǒng)計學意義(P0.05)。免疫組織化學檢測顯示,正常對照組(A組):視網膜各層排列整齊,VEGF見弱陽性表達,主要位于神經纖維層和內核層;高氧對照組(B組):視網膜組織結構紊亂,明顯水腫,可見大量VEGF表達,呈棕黃色,以內界膜,神經纖維層及新生血管內皮細胞表達最明顯。生理鹽水對照組(C組):視網膜各層組織水腫,細胞排列紊亂,VEGF分布在內界膜,神經纖維層及新生血管內皮細胞,,呈棕黃色,表達強;恩度干預組(D組):VEGF表達不明顯,各層均勻表達呈弱陽性。生理鹽水對照組(C組)分別與正常對照組(A組)比較,差異均有顯著性統(tǒng)計學意義(P0.01);高氧對照組(B組),生理鹽水對照組(C組)分別與恩度干預組(D組)比較,差異均有顯著性統(tǒng)計學意義(P0.01)。恩度干預組(D組)與正常對照組(A組)比較,差異具有統(tǒng)計學意義(P0.05)。 結論:腹腔內注射恩度可抑制小鼠模型中的視網膜新生血管形成,有望成為防治血管增生性視網膜病變的一種方法。
[Abstract]:Objective: To investigate the inhibitory effect of recombinant human endostatin (ENDOSTAR) on neovascularization in the oxygen-inducedretinopathy (OIR) model of hyperoxia induced mice
Methods: 60 C57BL/6J mice were randomly divided into normal control group (group A), hyperoxic control group (group B), normal saline control group (group C), and degree intervention group (group D), 15 rats in each group, except the normal control group (group A), the OIR model was established. The normal control group (A group), the hyperoxic control group (B group) mice did not do, the saline control group (C group) was in 12D. Intraperitoneal injection of 0.1mlNS, D group (Group) was given rh.ES injection 0.1ml by intraperitoneal injection at 12D, continuous injection of 5D, and rat age 17D, the number of neovascular endothelial nuclei breaking through the inner boundary membrane was counted by HE staining, and the expression of VEGF in the retina was detected by immunohistochemistry.
Results: the average number of nuclei of endothelial cells in the inner boundary membrane of each slice in the normal control group (group A) was (0.97 + 0.04), and the average number of nuclei of the endothelial cell nuclei of each slice breaking through the inner boundary membrane in the high oxygen control group (group B) was (24.30 + 1.73). The average of each slice in the saline control group (group C) broke through the inner boundary membrane. The number of cell nuclei of endothelial cells was (22.58 + 1.54). The number of nuclei of vascular endothelial cells that broke through the inner boundary membrane of the retina was (3.38 + 0.27). The number of nuclei in the neovascularization of the neovascularization in the hyperoxic control group (group B) and the saline control group (group C) was more than that of the normal control group (group A). Significant statistical significance (P < 0.01); the number of neovascular endothelial nuclei of the retinal breakthroughs in the retina was significantly reduced, compared with the hyperoxia control group (group B) and the saline control group (group C), the difference was statistically significant (P < 0.01). The difference between the degree intervention group (D group) and the normal control group (group A) was significantly different. Significant statistical significance (P0.05). The immunohistochemistry test showed that the normal control group (group A): the retina layers were arranged neatly, VEGF was weakly positive, mainly in the nerve fiber layer and the core layer, and the hyperoxia control group (group B): the retinal tissue structure was disorganized, the edema was obvious, and a large number of VEGF expressions were seen, with brown yellow, inner boundary membrane and nerve fiber. The expression of vascular endothelial cells in the vascular layer and neovascularization was most obvious. In the normal saline control group (group C), the tissues of the retina were edema, the cells were arranged in disorder, the VEGF distribution of the inner boundary membrane, the nerve fiber layer and the neovascular endothelial cells were brown and strong, and the expression of:VEGF was not obvious in the degree of grace intervention group (group D), and the homogeneous expression in each layer was weak positive. Compared with the normal control group (group A), the saline control group (group C) had significant statistical significance (P0.01), and in the hyperoxic control group (group B), the normal saline control group (group C) compared with the ENN intervention group (D group), the difference was statistically significant (P0.01). The difference was compared with the normal control group (D group) and the normal control group (A group). Statistical significance (P0.05).
Conclusion: intraperitoneal injection of Endostar inhibits the formation of retinal neovascularization in mouse models, and is expected to become a method for prevention and treatment of vascular proliferative retinopathy.
【學位授予單位】:石河子大學
【學位級別】:碩士
【學位授予年份】:2013
【分類號】:R774.1
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