發(fā)布時間：2018-05-30 08:03

  本文選題：LASIK + 周期性拉伸��； 參考：《太原理工大學》2012年碩士論文

【摘要】：準分子激光原位角膜磨鑲術(laser in situ keratomileusis,LASIK)是目前開展較為廣泛且應用于臨床效果良好的治療近視的角膜屈光手術。每年有近百萬人接受LASIK手術治療。LASIK手術主要通過改變角膜中央光學區(qū)的曲率,使屈光不正現(xiàn)象得以改善。此手術在角膜瓣下的基質層中進行,這樣做既保存了角膜上皮層和前彈力層的完整度,同時又可防止角膜上皮下霧狀渾濁(Haze)的發(fā)生。 LASIK術后角膜損傷修復過程是多因素參與的復雜過程。其中,力學環(huán)境的改變對LASIK術后角膜組織的修復是一個不可忽視的因素。LASIK術后角膜層變薄,在相同的眼內壓作用下,角膜所受的應力明顯增大,角膜成纖維細胞的力學環(huán)境發(fā)生了變化,可能影響其生長、增殖。堿性成纖維細胞生長因子(basic Fibroblast Growth Factor,bFGF)既能促進傷口愈合與受損組織的修復還能促使受損細胞發(fā)生相應的形態(tài)學改變。因此研究在體及離體不同力學環(huán)境下,角膜組織及角膜成纖維細胞中bFGF的表達與分布,以及力學刺激對兔角膜基質細胞增殖的影響,探究力學因素在LASIK術后角膜損傷修復中的作用,為臨床治療近視提供有力的理論依據(jù)和指導。 本文的主要研究內容及結論如下： 1.建立不同切削量的LASIK手術動物模型,使在體角膜處于不同力學環(huán)境中,采用免疫熒光染色觀察角膜上皮及基質層中bFGF表達；蘇木素-伊紅(HE)染色觀察角膜基質細胞增殖的變化。研究發(fā)現(xiàn)：bFGF在上皮及基質層中均有表達,但基質層中熒光強度總體高于上皮層,70%一周組基質層熒光強度明顯強于上皮層(P0.05)；與正常組相比,術后1周角膜細胞數(shù)量先減少,術后1月細胞數(shù)量增多。同一手術方式術后1月細胞數(shù)量顯著多于1周(P0.05)。 2.于LASIK術后1周和1月處死實驗動物提取組織蛋白,采用酶聯(lián)免疫吸附試驗(ELISA)方法檢鋇(?)bFGF含量。研究發(fā)現(xiàn)：LASIK術后1周,角膜基質床殘余30%組與對照組相比,bFGF含量顯著增高(P0.05)；LASIK術后1月,角膜基質床殘余30%組bFGF含量回落至正常水平,不同手術組之間無顯著性差異；不同時間點同一手術方式之間比較發(fā)現(xiàn),僅角膜基質床殘余30%組1周和1月有顯著差異(P0.05)。 3.體外采用FX-4000細胞力學柔性基底拉伸系統(tǒng),對原代提取的兔角膜成纖維細胞分別進行頻率為O.1Hz、拉伸幅度為5%、10%、15%的周期性機械拉伸,并于6h和24h收集細胞培養(yǎng)液上清,采用ELISA法檢(?)(?)bFGF的含量。研究發(fā)現(xiàn)：拉伸6h后,15%拉伸組bFGF含量較對照組顯著增高(P0.05),24h后顯著性降低(P0.05)。 實驗結果表明：在體不同切削量的LASIK手術后角膜基質細胞的增殖發(fā)生變化,bFGF的表達主要集中在基質層。LASIK術后角膜組織及周期性拉伸作用下角膜成纖維細胞bFGF早期表達的顯著上調,將有益于術后角膜組織的修復。
[Abstract]:Laser in situ keratomileusis (LASIKK) is a kind of corneal refractive surgery which is widely used in the treatment of myopia. Every year, nearly one million people are treated with LASIK. LASIK improves refractive errors mainly by changing the curvature of the central optical area of the cornea. This procedure is performed in the stromal layer below the corneal flap, which not only preserves the integrity of the epithelial layer and the anterior elastic layer of the cornea, but also prevents the occurrence of haze under the cornea. The repair process of corneal injury after LASIK is a complicated process involving many factors. Among them, the change of mechanical environment is an important factor in corneal tissue repair after LASIK. The corneal layer becomes thinner after LASIK. Under the same intraocular pressure, the stress of cornea increases obviously. The mechanical environment of corneal fibroblasts has changed, which may affect the growth and proliferation of corneal fibroblasts. Basic Fibroblast Growth FactorbFGFs can not only promote wound healing and repair of damaged tissues, but also induce the corresponding morphological changes of injured cells. Therefore, the expression and distribution of bFGF in corneal tissue and corneal fibroblasts in vitro and in vitro, and the effects of mechanical stimulation on the proliferation of rabbit corneal stromal cells were studied. To explore the role of mechanical factors in the repair of corneal injury after LASIK, and to provide theoretical basis and guidance for clinical treatment of myopia. The main contents and conclusions of this paper are as follows: 1. The animal model of LASIK operation with different cutting amount was established to observe the expression of bFGF in corneal epithelium and stromal layer by immunofluorescence staining and the proliferation of corneal stromal cells by hematoxylin-eosin) staining. It was found that the fluorescence intensity in the stromal layer was significantly higher than that in the epithelium 70% week group than that in the epithelial layer P0.05.The number of corneal cells decreased at the first week after operation, compared with the normal group, but the fluorescence intensity in the stromal layer was significantly higher than that in the control group. The number of cells increased 1 month after operation. The number of cells in one month after operation was significantly more than 1 week after operation (P 0.05). 2. After 1 week and 1 month after LASIK, the experimental animals were killed to extract the histone protein, and the content of bFGF was detected by Elisa. It was found that the content of bFGF in 30% group of residual corneal stroma bed was significantly higher than that in control group at 1 week after operation, and the content of bFGF in 30% group of residual corneal stroma bed decreased to the normal level one month after operation, but there was no significant difference between different groups. Compared with the same operation method at different time points, only the corneal stromal bed residual 30% group had significant difference in 1 week and 1 month (P 0.05). 3. In vitro, the primary rabbit corneal fibroblasts were subjected to mechanical stretching of 0.1 Hz with a tensile range of 5 1010% and 15% periodic mechanical stretching, and the supernatants were collected at 6 h and 24 h, respectively, using the FX-4000 cell mechanical flexible substrate stretching system, and the cultured rabbit corneal fibroblasts were collected at 6 h and 24 h respectively, and the cell culture medium supernatants were collected at 6 h and 24 h respectively. The content of bFGF was detected by ELISA. The results showed that the content of bFGF in the 15% stretch group was significantly higher than that in the control group after 6 hours of stretching. The results showed that the expression of bFGF in corneal stromal cells after LASIK surgery was mainly in the early stage of corneal fibroblast bFGF after stromal layer. LASIK and periodic stretching. A significant up-regulation of the expression, It will be beneficial to the repair of corneal tissue after operation.
【學位授予單位】：太原理工大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R779.63

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本文編號：1954554