發(fā)布時(shí)間：2018-05-01 08:21

  本文選題：角膜新生血管 + 穿透性角膜移植術(shù)��； 參考：《河北醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:我國(guó)現(xiàn)有角膜盲患者約300萬(wàn)人,且每年以10-20萬(wàn)的速度增長(zhǎng)。角膜移植術(shù)是治療角膜盲患者的最有效方法,術(shù)后角膜新生血管生長(zhǎng)仍是角膜移植手術(shù)失敗的主要因素。目前臨床上主要使用糖皮質(zhì)激素類藥物控制角膜移植術(shù)后角膜新生血管的生長(zhǎng)。糖皮質(zhì)激素類藥物的治療效果確鑿,但容易引起激素性青光眼、并發(fā)性白內(nèi)障等眼部并發(fā)癥。因此尋找一種既能有效控制角膜移植術(shù)后角膜新生血管的生長(zhǎng)又很少引起眼部并發(fā)癥的安全、高效的抗新生血管藥物顯得尤為重要。康柏西普是一種由我國(guó)自發(fā)研制的具有自主知識(shí)產(chǎn)權(quán)的新型抗血管內(nèi)皮生長(zhǎng)因子(vascular endothelial growth factor,VEGF)藥物,它已被廣泛使用于眼底新生血管的治療,國(guó)內(nèi)外尚無(wú)用于治療角膜移植術(shù)后新生血管的報(bào)道。本實(shí)驗(yàn)通過(guò)觀察康柏西普對(duì)兔穿透性角膜移植術(shù)(penetrating keratoplasty,PKP)術(shù)后的影響進(jìn)一步探索其在兔PKP術(shù)后新生血管生長(zhǎng)方面的治療作用。眾所周知,在血管化組織中應(yīng)用血管生長(zhǎng)抑制劑將影響組織創(chuàng)傷愈合過(guò)程。因此,康柏西普控制角膜新生血管生長(zhǎng)同時(shí)是否會(huì)阻礙角膜傷口的愈合是本實(shí)驗(yàn)需要關(guān)注的重要問(wèn)題。角膜傷口的愈合過(guò)程中,靜態(tài)的角膜基質(zhì)細(xì)胞被激活轉(zhuǎn)化為角膜成纖維細(xì)胞,損傷修復(fù)過(guò)程中部分角膜成纖維細(xì)胞形態(tài)功能進(jìn)一步改變轉(zhuǎn)化為肌成纖維細(xì)胞。肌成纖維細(xì)胞和平滑肌細(xì)胞有相同特征,具有收縮性,可以促進(jìn)損傷愈合。α-平滑肌肌動(dòng)蛋白(α-smooth muscle actin,α-SMA)是肌成纖維細(xì)胞的特異性細(xì)胞標(biāo)記物,它的測(cè)定能夠反應(yīng)角膜中肌成纖維細(xì)胞的水平,進(jìn)而推測(cè)角膜傷口的愈合程度。本實(shí)驗(yàn)旨在通過(guò)測(cè)定VEGF的表達(dá),初步探明康柏西普對(duì)兔PKP術(shù)后角膜新生血管的抑制作用;觀察α-SMA的變化推測(cè)角膜傷口修復(fù)過(guò)程以及康柏西普對(duì)傷口愈合的影響,從而指導(dǎo)人PKP術(shù)后新生血管的治療。方法:1分組:將48只新西蘭大白兔隨機(jī)分為4組,每組12只。分別為:對(duì)照組、實(shí)驗(yàn)1組、實(shí)驗(yàn)2組和實(shí)驗(yàn)3組。對(duì)照組:常規(guī)點(diǎn)眼,即兔PKP造模后妥布霉素滴眼液點(diǎn)術(shù)眼4/日;實(shí)驗(yàn)1組:常規(guī)點(diǎn)眼,同時(shí)加用0.01%康柏西普點(diǎn)術(shù)眼4/日;實(shí)驗(yàn)2組:常規(guī)點(diǎn)眼,同時(shí)加用0.1%康柏西普點(diǎn)術(shù)眼4/日;實(shí)驗(yàn)3組:常規(guī)點(diǎn)眼,同時(shí)開(kāi)始0.1%康柏西普0.1ml結(jié)膜下注射1/周。2制備兔PKP模型:7.5mm環(huán)鉆制備植片;7.25mm環(huán)鉆鉆切,15°前房穿刺刀鉆痕處穿刺,角膜剪剪下角膜片制成植床;10-0尼龍線先于12:00、6:00、9:00、3:00各縫合1針,再于每象限加縫2針,縫合植片。48只兔右眼均按照以上步驟制備PKP模型。3實(shí)時(shí)熒光定量聚合酶鏈?zhǔn)椒磻?yīng)(Real time-polymerase chain reaction,q PCR)術(shù)后第2周、4周、6周、8周各時(shí)間點(diǎn)每組處死3只兔,取材后采用q PCR方法檢測(cè)各時(shí)間點(diǎn)角膜VEGF和α-SMA的表達(dá)情況。4統(tǒng)計(jì):應(yīng)用SPSS21.0軟件進(jìn)行統(tǒng)計(jì)學(xué)分析。結(jié)果:1 VEGF的表達(dá):對(duì)照組和各個(gè)實(shí)驗(yàn)組VEGF的表達(dá)均隨時(shí)間推移而先增加后減少,且第4周時(shí)VEGF的表達(dá)量最高;所有時(shí)間點(diǎn)各實(shí)驗(yàn)組與對(duì)照組比較,VEGF表達(dá)均降低且有統(tǒng)計(jì)學(xué)意義(P0.05);實(shí)驗(yàn)1組和實(shí)驗(yàn)2組比較,各時(shí)間點(diǎn)VEGF表達(dá)量均無(wú)統(tǒng)計(jì)學(xué)差異(P0.05),實(shí)驗(yàn)3組所有時(shí)間點(diǎn)VEGF表達(dá)較實(shí)驗(yàn)2組低,降低程度無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。2α-SMA的表達(dá):隨時(shí)間進(jìn)展,所有組α-SMA表達(dá)均呈上升趨勢(shì),所有組內(nèi)任意時(shí)間點(diǎn)兩兩比較均有統(tǒng)計(jì)學(xué)差異(P0.05);所有實(shí)驗(yàn)組4個(gè)時(shí)間點(diǎn)α-SMA表達(dá)均低于對(duì)照組,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);實(shí)驗(yàn)1組和實(shí)驗(yàn)2組比較,實(shí)驗(yàn)2組和實(shí)驗(yàn)3組比較,各時(shí)間點(diǎn)α-SMA表達(dá)均無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。結(jié)論:1康柏西普可以抑制兔穿透性角膜移植術(shù)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術(shù)后角膜新生血管生成的同時(shí)不影響角膜傷口的愈合。3不同給藥濃度對(duì)兔穿透性角膜移植術(shù)后角膜新生血管的作用無(wú)差別,不同給藥方式對(duì)新生血管的生長(zhǎng)抑制作用還有待后續(xù)研究進(jìn)一步證實(shí)。
[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.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R779.65

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本文編號(hào)：1828509