發(fā)布時間：2018-11-03 07:02

【摘要】： 目的：利用兔眼青光眼濾過術(shù)(小梁切除)模型,予濾過泡旁球結(jié)膜下注射三種不同劑量的Avastin,探討Avastin在抑制青光眼濾過術(shù)后瘢痕化方面的療效,結(jié)膜下注射的時機(jī)和劑量,同時對Avastin結(jié)膜下注射是否對術(shù)區(qū)組織產(chǎn)生毒性反應(yīng)作初步研究。 方法：眼壓正常的新西蘭白兔50只50眼隨機(jī)分為三個不同濃度的Avastin實(shí)驗(yàn)組,即0.5mgAvastin實(shí)驗(yàn)組(A組)、1.0mgAvastin實(shí)驗(yàn)組(B組)、2.0mgAvastin實(shí)驗(yàn)組(C組),以及絲裂霉素C(MMC)對照組和空白對照組,每組10眼。對兔眼行常規(guī)小梁切除術(shù),A、B、C實(shí)驗(yàn)組于術(shù)畢及術(shù)后第3、7天濾過泡旁球結(jié)膜下分別注射6.25mg／m1、12.50mg／ml、25.0mg/ml Avastin 0.08ml,即0.5、1.0、2.0mg；MMC對照組術(shù)中筋膜囊、鞏膜瓣下放置浸濕0.2mg／m1 MMC棉片；空白對照組于術(shù)畢及術(shù)后第3、7天濾過泡旁球結(jié)膜下注射0.9%氯化鈉注射液0.08ml。術(shù)后裂隙燈顯微鏡觀察濾過泡情況,并行活體共焦顯微鏡檢查；測量眼壓；于術(shù)后第7、14天,每組摘除眼球5只,常規(guī)HE染色,于高倍鏡下計算濾過泡腔面及形成區(qū)面積占結(jié)膜面積的平均百分比,Tunel法檢測細(xì)胞凋亡情況,同時ELISA法測定房水及組織勻漿TGF-β1,bFGF含量。 結(jié)果：(1)裂隙燈顯微鏡檢查：術(shù)后第7天,A、B、C實(shí)驗(yàn)組及MMC對照組濾過泡隆起彌散,表面血管稀疏；空白對照組術(shù)后第3天濾過泡開始局限化,至第7天濾過泡周圍怒張粗大的血管形成。術(shù)后第14天,C實(shí)驗(yàn)組及MMC對照組濾過泡扁平彌散,表面血管稀疏纖細(xì)；B實(shí)驗(yàn)組濾過泡扁平彌散,表面血管較7天時密集；A實(shí)驗(yàn)組濾過泡血管包裹局限；空白對照組濾過泡瘢痕化、表面血管濃密。(2)眼壓：術(shù)后14天內(nèi)各組眼壓均隨時間呈線性增長,但增長趨勢無差別,且不同時間點(diǎn)各組間眼壓無差異。(3)活體共焦顯微鏡檢查：術(shù)后第7天,A、B、C實(shí)驗(yàn)組及MMC對照組較空白對照組濾過泡囊腔結(jié)構(gòu)明顯,纖維組織疏松；MMC對照組組織層間見大量炎癥細(xì)胞浸潤。術(shù)后第14天,C實(shí)驗(yàn)組及MMC對照組仍可見明顯濾過泡囊腔結(jié)構(gòu),纖維組織纖細(xì)疏松；B實(shí)驗(yàn)組殘留少許濾過泡結(jié)構(gòu)；其余各組纖維組織致密,縱橫交錯,血管粗大呈螺旋狀。各組角膜結(jié)構(gòu)均未見異常。(4)HE染色：術(shù)后第7天,各組可見明顯濾過泡結(jié)構(gòu)。術(shù)后第14天,C實(shí)驗(yàn)組及MMC對照組可見明顯濾過泡結(jié)構(gòu)；B實(shí)驗(yàn)組見少量微小的濾過泡；A實(shí)驗(yàn)組及空白對照組濾過泡消失,纖維組織增生明顯。采用單因素方差分析顯示：術(shù)后第7天,各組濾過泡腔面及形成區(qū)面積與結(jié)膜面積的平均百分比無差別；術(shù)后第14天,C實(shí)驗(yàn)組及MMC對照組濾過泡腔面占結(jié)膜的平均面積百分比高于A、B實(shí)驗(yàn)組及空白對照組,B實(shí)驗(yàn)組濾過泡腔面在結(jié)膜的平均面積百分比高于A實(shí)驗(yàn)組及空白對照組,而C實(shí)驗(yàn)組與MMC對照組間及A實(shí)驗(yàn)組與空白對照組間面積百分比無差別。C實(shí)驗(yàn)組及MMC對照組濾過泡形成區(qū)占結(jié)膜平均面積百分比高于A、B實(shí)驗(yàn)組及空白對照組；C實(shí)驗(yàn)組與MMC對照組間及A、B實(shí)驗(yàn)組與空白對照組間無差別。(5)ELISA法測定TGF-β1、bFGF含量：術(shù)后第7天及第14天,C實(shí)驗(yàn)組及MMC對照組房水、術(shù)區(qū)組織中TGF-β1、bFGF含量均低于A、B實(shí)驗(yàn)組及空白對照組；B實(shí)驗(yàn)組TGF-β1、bFGF含量低于A實(shí)驗(yàn)組及空白對照組；C實(shí)驗(yàn)組與MMC對照組組間及A實(shí)驗(yàn)組與空白對照組間TGF-β1、bFGF含量無差別。(6)Tunel細(xì)胞凋亡檢測：術(shù)后第7天,各實(shí)驗(yàn)組及空白對照組術(shù)區(qū)組織中偶見強(qiáng)熒光的凋亡細(xì)胞,MMC對照組見大量凋亡細(xì)胞。術(shù)后第14天,A、B實(shí)驗(yàn)組及空白對照組偶見凋亡細(xì)胞；C實(shí)驗(yàn)組結(jié)膜上皮下、筋膜囊組織見少量凋亡細(xì)胞；MMC對照組仍見大量凋亡細(xì)胞。采用單因素方差分析顯示：術(shù)后第7、14天,MMC對照組凋亡細(xì)胞均最多,高于各實(shí)驗(yàn)組及空白對照組；空白對照組與實(shí)驗(yàn)組組向凋亡細(xì)胞數(shù)無差別 結(jié)論：術(shù)后早期濾過泡旁球結(jié)膜下注射2.0mg及1.0mgAvastin可有效抑制濾過泡新生血管的形成,有助于緩解濾過泡的瘢痕化,促進(jìn)功能性濾過泡的形成與維持,且安全性較好。
[Abstract]:Objective: To study the therapeutic effect of Avastin in the treatment of glaucoma after filtration, and to explore the time and dosage of avastin in the treatment of glaucoma after filtration. At the same time, the toxicity of Avastin subconjunctival injection was studied. Methods: 50 eyes of New Zealand white rabbits with normal intraocular pressure were randomly divided into three experimental groups with different concentrations, namely 0. 5mgAvastin experimental group (group A), 1. 0mgAvastin experimental group (group B), 2. 0mgAvastin experimental group (group C), and MMC (MMC) control group and blank control group. 10 eyes of group A, B and C were respectively injected with 6. 25mg/ ml, 12.50mg/ ml, 25. 0mg/ ml Avastin 0. 08ml, that is, 0. 5, 1. 0, 2. 0mg. MC cotton tablets, blank control group were injected with 0.9% sodium chloride injection at the 3 rd and 7 th day after operation, and 0.9% sodium chloride injection was injected. 08ml. After operation, the slit lamp microscope was used to observe the filter bubble condition, and the confocal microscopy was performed in parallel; the intraocular pressure was measured; after 7 and 14 days after operation, 5 eyes were removed from each group, conventional HE staining was performed, and the filtration bubble cavity surface and the area occupied by the formation area accounted for the level of the conjunctival area at high magnification. Both percentage and Tunel method were used to detect the apoptosis of the cells. GF content. Results: (1) Slit lamp microscope examination: After 7 days after operation, the alveolar ridge was dispersed in group A, B, C and MMC control group, and the surface blood vessel was sparse; 3 days after operation of blank control group, the bleb began to localize until the 7th day after filtration bubble. During the 14th day after operation, the experimental group and MMC control group filtered the flat dispersion, the surface blood vessels were sparse and thin, the experimental group was filtered with flat dispersion, and the surface vessels were dense at 7 days. (2) Intraocular pressure: The intraocular pressure in each group increased linearly with time within 14 days after operation, but there was no difference in the growth trend and different time. There was no difference in intraocular pressure in each group. (3) In-vivo confocal microscope examination: group A, B, C and MMC in control group were compared with blank control group, the structure of alveolar cavity was obvious, the fibrous tissue was loose, and the tissue layers of MMC control group A lot of inflammatory cell infiltration was found. After the 14th day of operation, the experimental group and MMC control group still showed a clear filter vesicle cavity structure, and the fibrous tissue was fine and loose; B experimental group had a little residual filtration cell structure; the other groups of fibrous tissue were dense and dry. and the blood vessels are in a spiral shape. The corneal structure was abnormal. (4) HE staining: 7 days after operation, each group The results showed that the bleb structure was found in the experimental group and the MMC control group after the 14th day of operation. The experimental group showed a small amount of small filtration bubble; the experimental group and the blank control group were filtered out to disappear. The results of single-factor analysis of variance showed that there was no difference between the surface area of each group and the area of formation area and the average percentage of conjunctival area after 7 days of operation. The percentage of soaking area was higher than that of group A, group B and blank control group, and the percentage of average area of filter cell in group B was higher than that of group A and blank control group, and between control group and MMC control group and control group A and control group B. The percentage of interconjunctival area in experimental group and MMC control group was higher than that of group A, group B and blank control group, and the experimental group and MMC control group and group A and B were compared with control group. There was no difference in the blank control group. (5) The content of bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, bFGF, The content of bFGF in experimental group was lower than that of group A and blank control group, and between experimental group and MMC control group and between group A and blank control group. There was no difference in bFGF content. (6) Apoptosis of Tunel cells: In the 7th day after operation, the apoptotic cells of strong fluorescence were occasionally observed in each experimental group and the blank control group. In group A, group B and blank control group, apoptotic cells were occasionally seen in group A, group B and blank control group after operation. A large number of apoptotic cells were still seen in the group. The single factor analysis of variance showed that the apoptotic cells in the MMC control group were most in the 7th and 14th days after operation, higher than those in each experimental group and the control group, and the blank control group and the experimental group. group's orientation There was no difference in the number of apoptotic cells: 2. 0mg and 1. 0mgAvasin could effectively inhibit the formation of filter bubble neovascularization at the early stage after operation.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R779.6

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