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目的制作大鼠角膜百草枯化学烧伤模型,观察在角膜新生血管(CNV)形成过程中核转录因子-κβ(NF-κβ)和血管内皮生长因子(VEGF)的动态表达,研究百草枯所致角膜烧伤后的病理变化以及CNV形成的机制。方法将30只健康Wistar大鼠随机分成两组:A组(空白对照组)5只,B组(实验对照组)25只。制作大鼠角膜百草枯烧伤模型后,两组每天氯霉素滴眼液点眼预防感染。术后每天观察大鼠角膜的炎症反应及CNV生长情况。在术后第1、4、7、14及21天,用裂隙灯拍照记录该组所有大鼠角膜的改变,并处死,取全角膜,行HE染色和免疫组织化学染色,观察角膜炎性反应及CNV的生长情况,同时检测NF-κβ和VEGF的动态表达,并进行光密度值测定和分析。结果 1HE染色:术后第4天:B组角膜上皮坏死,基质紊乱,大量炎细胞浸润;术后第7天:B组角膜上皮增厚,基质中出现新生血管,可见成纤维细胞增殖;术后14d:B组角膜有大量新生血管,胶原纤维排列紊乱,仍有炎细胞浸润;术后21d:B组角膜上皮代偿性增生,基质中炎细胞和新生血管均减少。2免疫组织化学染色:A组正常大鼠角膜有微弱的NF-κβ和VEGF表达。B组大鼠角膜组织中NF-κβ和VEGF表达均上调。NF-κβ于第4天表达最明显,第7天逐渐减弱,第14天后逐渐降至正常水平,VEGF于第7天表达最明显,第14天后明显减弱。结论大鼠角膜百草枯烧伤后,NF-κβ和VEGF的表达明显增加,NF-κβ和VEGF的高表达可能是角膜烧伤后形成CNV的重要因素。
OBJECTIVE: To establish a rat model of corneal chemical paraquat burn injury and to observe the dynamic expression of nuclear factor-κβ (NF-κβ) and vascular endothelial growth factor (VEGF) in corneal neovascularization (CNV) Post-pathological changes and the mechanism of CNV formation. Methods Thirty healthy Wistar rats were randomly divided into two groups: A group (blank control group) 5, B group (experimental control group) 25. After the rat cornea paraquat burn model was made, two groups of chloramphenicol eye drops were spotted every day to prevent infection. The corneal inflammation and CNV growth were observed daily after operation. On the 1st, 4th, 7th, 14th and 21st day after the operation, the corneal changes of all the rats in this group were photographed by slit lamp and sacrificed. The whole cornea was taken and stained with HE and immunohistochemistry to observe the corneal inflammatory reaction And the growth of CNV. At the same time, the dynamic expression of NF-κβ and VEGF were detected, and the optical density was measured and analyzed. Results 1HE staining: corneal epithelial necrosis, stromal disorders and a large number of inflammatory cell infiltration in group B at day 4 postoperatively. On the 7th day after operation, the corneal epithelium in group B was thickened and neovascularization appeared in the stroma, and fibroblast proliferation was observed After 14 days, there were a large number of corneal neovascularization in Group B, with disorganized collagen fibers and infiltration of inflammatory cells. On day 21 postoperatively, corneal epithelium in Group B compensated hyperplasia and inflammatory cells and neovascularization in the matrix decreased. Immunohistochemical staining: A group of normal rat cornea had weak NF-κβ and VEGF expression. In group B, the expression of NF-κβ and VEGF were up-regulated in corneal tissue. The expression of NF-|ÊB was most obvious on the 4th day, gradually weakened on the 7th day, gradually decreased to the normal level on the 14th day, and the expression of NF-|Ê|Â was the most obvious on the 7th day and significantly decreased on the 14th day. Conclusion The expression of NF-κβ and VEGF is significantly increased after the corneal bleaching injury in rat cornea. The high expression of NF-κβ and VEGF may be an important factor in the formation of CNV after corneal burn.