康柏西普对兔穿透性角膜移植术后角膜新生血管的影响

发布时间：2018-05-01 08:21

本文选题：角膜新生血管 + 穿透性角膜移植术　；参考：《河北医科大学》2017年硕士论文

【摘要】：目的:我国现有角膜盲患者约300万人,且每年以10-20万的速度增长。角膜移植术是治疗角膜盲患者的最有效方法,术后角膜新生血管生长仍是角膜移植手术失败的主要因素。目前临床上主要使用糖皮质激素类药物控制角膜移植术后角膜新生血管的生长。糖皮质激素类药物的治疗效果确凿,但容易引起激素性青光眼、并发性白内障等眼部并发症。因此寻找一种既能有效控制角膜移植术后角膜新生血管的生长又很少引起眼部并发症的安全、高效的抗新生血管药物显得尤为重要。康柏西普是一种由我国自发研制的具有自主知识产权的新型抗血管内皮生长因子(vascular endothelial growth factor,VEGF)药物,它已被广泛使用于眼底新生血管的治疗,国内外尚无用于治疗角膜移植术后新生血管的报道。本实验通过观察康柏西普对兔穿透性角膜移植术(penetrating keratoplasty,PKP)术后的影响进一步探索其在兔PKP术后新生血管生长方面的治疗作用。众所周知,在血管化组织中应用血管生长抑制剂将影响组织创伤愈合过程。因此,康柏西普控制角膜新生血管生长同时是否会阻碍角膜伤口的愈合是本实验需要关注的重要问题。角膜伤口的愈合过程中,静态的角膜基质细胞被激活转化为角膜成纤维细胞,损伤修复过程中部分角膜成纤维细胞形态功能进一步改变转化为肌成纤维细胞。肌成纤维细胞和平滑肌细胞有相同特征,具有收缩性,可以促进损伤愈合。α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)是肌成纤维细胞的特异性细胞标记物,它的测定能够反应角膜中肌成纤维细胞的水平,进而推测角膜伤口的愈合程度。本实验旨在通过测定VEGF的表达,初步探明康柏西普对兔PKP术后角膜新生血管的抑制作用;观察α-SMA的变化推测角膜伤口修复过程以及康柏西普对伤口愈合的影响,从而指导人PKP术后新生血管的治疗。方法:1分组:将48只新西兰大白兔随机分为4组,每组12只。分别为:对照组、实验1组、实验2组和实验3组。对照组:常规点眼,即兔PKP造模后妥布霉素滴眼液点术眼4/日;实验1组:常规点眼,同时加用0.01%康柏西普点术眼4/日;实验2组:常规点眼,同时加用0.1%康柏西普点术眼4/日;实验3组:常规点眼,同时开始0.1%康柏西普0.1ml结膜下注射1/周。2制备兔PKP模型:7.5mm环钻制备植片;7.25mm环钻钻切,15°前房穿刺刀钻痕处穿刺,角膜剪剪下角膜片制成植床;10-0尼龙线先于12:00、6:00、9:00、3:00各缝合1针,再于每象限加缝2针,缝合植片。48只兔右眼均按照以上步骤制备PKP模型。3实时荧光定量聚合酶链式反应(Real time-polymerase chain reaction,q PCR)术后第2周、4周、6周、8周各时间点每组处死3只兔,取材后采用q PCR方法检测各时间点角膜VEGF和α-SMA的表达情况。4统计:应用SPSS21.0软件进行统计学分析。结果:1 VEGF的表达:对照组和各个实验组VEGF的表达均随时间推移而先增加后减少,且第4周时VEGF的表达量最高;所有时间点各实验组与对照组比较,VEGF表达均降低且有统计学意义(P0.05);实验1组和实验2组比较,各时间点VEGF表达量均无统计学差异(P0.05),实验3组所有时间点VEGF表达较实验2组低,降低程度无统计学差异(P0.05)。2α-SMA的表达:随时间进展,所有组α-SMA表达均呈上升趋势,所有组内任意时间点两两比较均有统计学差异(P0.05);所有实验组4个时间点α-SMA表达均低于对照组,差异无统计学意义(P0.05);实验1组和实验2组比较,实验2组和实验3组比较,各时间点α-SMA表达均无统计学差异(P0.05)。结论:1康柏西普可以抑制兔穿透性角膜移植术后角膜新生血管的生成。2康柏西普在抑制兔穿透性角膜移植术后角膜新生血管生成的同时不影响角膜伤口的愈合。3不同给药浓度对兔穿透性角膜移植术后角膜新生血管的作用无差别,不同给药方式对新生血管的生长抑制作用还有待后续研究进一步证实。
[Abstract]:Objective: there are about 3 million people with corneal blindness in China, and increase at a rate of 10-20 000 per year. Corneal transplantation is the most effective method for the treatment of corneal blindness. Corneal neovascularization is still the main factor for the failure of corneal transplantation. The growth of neovascularization. The therapeutic effect of glucocorticoids is conclusive, but it is easy to cause ocular complications such as hormonal glaucoma and complicated cataract. Therefore, it is safe to find a kind of corneal neovascularization which can not only effectively control the growth of corneal neovascularization after corneal transplantation, but also more effective anti neovascularization drugs. Compacept is a new anti vascular endothelial growth factor (vascular endothelial growth factor, VEGF) drug, which is spontaneously developed by our country. It has been widely used in the treatment of neovascularization of the fundus. There is no report on the treatment of neovascularization after the treatment of angular membrane transplantation. The effect of CORB cept on penetrating keratoplasty (PKP) after the rabbit's penetrating keratoplasty (PKP) is further explored. It is known that the application of vascular growth inhibitors in vascular tissue will affect the healing process of tissue injury. Therefore, Corp cept controls corneal neoplastic. During the healing process of corneal wound, the static corneal stroma cells are activated and converted into corneal fibroblasts, and the morphological work of some corneal fibroblasts can be further transformed into myofibroblast during the repair process. Fibroblasts and smooth muscle cells have the same characteristics, they have contractility and can promote the healing of injury. Alpha smooth muscle actin (alpha -smooth muscle actin, alpha -SMA) is a specific cell marker of myofibroblast. Its determination can reflect the level of myofibroblast in the cornea, and then speculate the healing degree of corneal wound. The experiment aims to determine the inhibitory effect of compaulcept on corneal neovascularization after PKP operation in rabbits by measuring the expression of VEGF, and to observe the changes of the corneal wound healing process and the effect of conb cept on the wound healing by observing the changes of alpha -SMA, and thus guiding the treatment of newborn blood vessels after PKP. Method: 1 groups: 48 New Zealand white rabbits were randomly divided into groups. 4 groups were divided into 4 groups: the control group, the experiment 1 groups, the experimental 2 groups and the experiment 3 groups. The control group: the normal eye, that is, the 4/ day of the Tobramycin Eye Drops point operation in the rabbit after the model of the rabbit model, the 1 group: the routine eye, and the 4/ day of 0.01% compaetcept; the experiment 2: the conventional eyes, and 0.1% compacept point operation 4/ day; Experiment 3 Group: routine eyes, and starting 0.1% CONP cept 0.1ml conjunctiva for 1/ week.2 to prepare rabbit PKP model: 7.5mm ring drill; 7.25mm ring drill cut, 15 degree anterior chamber piercing piercing scar, cornea cut angle diaphragm to implant bed; 10-0 nylon line before 12:00,6:00,9: 00,3:00, 1 stitches, 2 stitches per quadrant and suture implant.4 The 8 rabbit right eyes were prepared by the above steps to prepare PKP model.3 real-time quantitative polymerase chain reaction (Real time-polymerase chain reaction, Q PCR) for second weeks, 4 weeks, 6 weeks, and 8 weeks at each time point in each group of 3 rabbits, and then used Q PCR to detect the expression of VEGF and alpha -SMA. Results: 1 VEGF expression: the expression of VEGF in the control group and the experimental group increased first and then decreased with the passage of time, and the expression of VEGF was the highest at fourth weeks. All the experimental groups were compared with the control group at all time points, and the expression of VEGF decreased and had statistical significance (P0.05); the 1 groups and the experimental groups were compared at all time. There was no statistical difference in the expression of point VEGF (P0.05). The expression of VEGF in all time points in the 3 groups was lower than that in the experimental 2 groups, and the degree of reduction was not statistically significant (P0.05).2 alpha -SMA expression. All groups of alpha -SMA expression increased with time, and there was a statistically significant difference between all groups at any time point 22 (P0.05), and all the experimental groups were 4. The expression of inter point alpha -SMA was lower than that of the control group (P0.05). Compared with the experimental group 1 and the experiment 2 groups, the expression of alpha -SMA in the 2 groups and the experimental 3 groups was no significant difference (P0.05). Conclusion: 1 compacp can inhibit the formation of corneal neovascularization after penetrating keratoplasty in rabbits,.2 compacp is inhibited in rabbits Corneal neovascularization after penetrating keratoplasty does not affect the healing of corneal wound at the same time. There is no difference in the effect of different doses of.3 on the corneal neovascularization after penetrating keratoplasty in rabbits. The inhibition of the growth of new blood vessels by different drug delivery methods remains to be further confirmed.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R779.65

【参考文献】