本文選題：高度近視 + 后鞏膜加固術(shù)　； 參考：《太原理工大學(xué)》2011年博士論文

【摘要】：近視是常見的眼部疾病,有屈光性近視與進行性近視之分。其中,進行性近視又稱高度近視或病理性近視,在人群中患病率很高,可發(fā)生很多嚴(yán)重并發(fā)癥,是常見的致盲原因之一。后鞏膜加固術(shù)能夠提高后極部鞏膜的強度,阻止眼軸進一步延長,最終阻止視功能進一步惡化,是治療高度近視較為有效的一種方法。 有關(guān)后鞏膜加固術(shù)的療效說法不一,有必要對后鞏膜加固術(shù)的治療機理進行更深入、全面地研究。本文對建立的新西蘭大白兔高度近視眼動物模型實施后鞏膜加固手術(shù),術(shù)后不同時期獲取加固區(qū)鞏膜組織與鞏膜成纖維細(xì)胞,研究后鞏膜加固術(shù)后鞏膜及鞏膜成纖維細(xì)胞蛋白的表達(dá)情況、鞏膜成纖維細(xì)胞的增殖活性和粘彈性、鞏膜成纖維細(xì)胞對力學(xué)刺激的響應(yīng),從力學(xué)-細(xì)胞生物學(xué)角度解釋后鞏膜加固術(shù)的治療機理,這將為該手術(shù)在臨床上的選擇實施和推廣提供理論指導(dǎo)。 本文的主要研究內(nèi)容及結(jié)論如下： 1.選取3周齡未脫離母乳喂養(yǎng)的新西蘭大白兔40只,隨機、單眼進行眼瞼縫合手術(shù)誘導(dǎo)建立高度近視眼模型,另一眼為對照眼。眼瞼縫合后60天,打開眼臉,檢驗屈光度和測量眼軸長度。經(jīng)過誘導(dǎo)后,實驗眼與對照眼屈光度比較,誘導(dǎo)出1.80±0.72D的相對近視,眼軸相對延長了0.45±0.29mm,差異具有統(tǒng)計學(xué)意義(P0.05),表明動物模型建立成功。 2.對高度近視眼模型實施后鞏膜加固手術(shù),術(shù)后3個月和6個月提取加固區(qū)鞏膜組織和正常鞏膜組織,制備組織切片,通過免疫組化方法檢測術(shù)后鞏膜基質(zhì)金屬蛋白酶-2(MMP-2)及其特異性組織抑制劑(TIMP-2)、轉(zhuǎn)化生長因子-β1 (TGF-β1)及堿性成纖維細(xì)胞生長因子(bFGF)的表達(dá)。檢測結(jié)果顯示：后鞏膜加固術(shù)后鞏膜組織MMP-2表達(dá)減少,而TIMP-2、TGF-β1及bFGF的表達(dá)增加。這些變化有利于鞏膜成纖維細(xì)胞的增殖,抑制鞏膜基質(zhì)蛋白降解,促進胞外基質(zhì)合成,使鞏膜組織增厚。 3.組織塊培養(yǎng)法提取并培養(yǎng)正常鞏膜組織、術(shù)后3個月、6個月原鞏膜組織與融合區(qū)組織鞏膜成纖維細(xì)胞,用免疫細(xì)胞化學(xué)法對獲取的細(xì)胞進行鑒定,結(jié)果顯示Vimentin染色陽性,Desmin染色陰性,keratin染色陰性,S-100染色陰性,證實所獲得的細(xì)胞為鞏膜成纖維細(xì)胞。 采用ATP熒光檢測法檢測各組細(xì)胞的增殖活性；免疫細(xì)胞化學(xué)及酶聯(lián)免疫吸附試驗(ELISA)的方法聯(lián)合檢測各組鞏膜成纖維細(xì)胞MMP-2、TIMP-2、TGF-β1及bFGF的表達(dá)；微管吸吮技術(shù)檢測各組鞏膜成纖維細(xì)胞的粘彈性參數(shù)(瞬時模量E0、平衡模量E∞和表觀粘性μ)。研究發(fā)現(xiàn)： (1)后鞏膜加固術(shù)后3個月和6個月鞏膜成纖維細(xì)胞的增殖活性均比正常鞏膜組織有明顯提高(P0.05),且術(shù)后融合區(qū)鞏膜成纖維細(xì)胞的增殖活性明顯高于原鞏膜組織(P0.05)。 (2)正常鞏膜組織、術(shù)后3個月和6個月原鞏膜組織的鞏膜成纖維細(xì)胞之間MMP-2的表達(dá)無明顯差異(P0.05),術(shù)后3個月和6個月融合區(qū)鞏膜成纖維細(xì)胞MMP-2的表達(dá)明顯低于術(shù)后原鞏膜組織和正常鞏膜組織(P0.05)；術(shù)后3個月和6個月原鞏膜組織與融合區(qū)鞏膜成纖維細(xì)胞TIMP-2、TGF-β1的表達(dá)要明顯高于正常鞏膜組織(P0.05)；術(shù)后3個月和6個月原鞏膜組織與融合區(qū)鞏膜成纖維細(xì)胞bFGF的表達(dá)要明顯高于正常鞏膜組織(P0.05),術(shù)后3個月和6個月融合區(qū)鞏膜成纖維細(xì)胞bFGF的表達(dá)明顯高于原鞏膜組織(P0.05)；術(shù)后3個月和6個月原鞏膜組織之間、術(shù)后3個月和6個月融合區(qū)之間鞏膜成纖維細(xì)胞MMP-2、TIMP-2、TGF-β1及bFGF的表達(dá)均無明顯差異(P0.05)。 (3)正常鞏膜組織與術(shù)后3個月、6個月原鞏膜組織鞏膜成纖維細(xì)胞之間的各項粘彈性參數(shù)(E0、E∞和μ)無明顯差異(P0.05),術(shù)后3個月、6個月融合區(qū)鞏膜成纖維細(xì)胞的各項粘彈性參數(shù)(E0、E∞和μ)均小于術(shù)后原鞏膜組織和正常鞏膜組織(P0.05)。 實驗結(jié)果表明：后鞏膜加固術(shù)后加固區(qū)鞏膜成纖維細(xì)胞的增殖活性提高,MMP-2表達(dá)減少,而TIMP-2、TGF-β1及bFGF的表達(dá)增加,與免疫組化結(jié)果是一致的,這些變化有利于鞏膜成纖維細(xì)胞外基質(zhì)的沉積和鞏膜組織增厚。后鞏膜加固術(shù)后原鞏膜組織鞏膜成纖維細(xì)胞的各項粘彈性參數(shù)(E0、E∞和μ)沒有發(fā)生變化,而融合區(qū)新生鞏膜成纖維細(xì)胞的各項粘彈性參數(shù)(E0、E∞和μ)較低。 4.取正常鞏膜組織,術(shù)后6個月原鞏膜組織、術(shù)后6個月融合區(qū)組織的鞏膜成纖維細(xì)胞,采用FX-4000系統(tǒng)施加0.1Hz、幅度為3%和6%的周期性拉伸,拉伸48h后收集細(xì)胞及細(xì)胞上清液,檢測各組細(xì)胞的增殖活性、MMP-2、TIMP-2、TGF-β1及bFGF的表達(dá)和粘彈性。研究發(fā)現(xiàn)： (1)經(jīng)過拉伸培養(yǎng)之后正常鞏膜組織和術(shù)后6個月原鞏膜組織鞏膜成纖維細(xì)胞的增殖活性與各自靜態(tài)對照組比較有明顯提高(P0.05),正常鞏膜組織6%拉伸組細(xì)胞的增殖活性明顯高于3%拉伸組(P0.05),術(shù)后6個月融合區(qū)鞏膜成纖維細(xì)胞在經(jīng)過力學(xué)環(huán)境培養(yǎng)后增殖活性與靜態(tài)組比較無統(tǒng)計學(xué)差異(P0.05)。 (2)正常鞏膜組織和術(shù)后6個月鞏膜組織的鞏膜成纖維細(xì)胞在經(jīng)過力學(xué)環(huán)境培養(yǎng)后,MMP-2的表達(dá)減少,TIMP-2的表達(dá)增加(P0.05),而術(shù)后6個月融合區(qū)鞏膜成纖維細(xì)胞MMP-2、TIMP-2的表達(dá)與靜態(tài)組比較無明顯差異(P0.05)；三組鞏膜成纖維細(xì)胞在經(jīng)過力學(xué)環(huán)境培養(yǎng)后,TGF-β1的表達(dá)均增加(P0.05)；正常鞏膜組織和術(shù)后6個月鞏膜組織的鞏膜成纖維細(xì)胞在經(jīng)過力學(xué)環(huán)境培養(yǎng)后,bFGF的表達(dá)增加(P0.05),術(shù)后6個月融合區(qū)鞏膜成纖維細(xì)胞bFGF的表達(dá)與靜態(tài)組比較無明顯差異(P0.05)。 (3)正常鞏膜組織與術(shù)后6個月原鞏膜組織鞏膜成纖維細(xì)胞在經(jīng)過力學(xué)環(huán)境培養(yǎng)后,細(xì)胞的粘彈性參數(shù)(E0、E∞和μ)與各自靜態(tài)培養(yǎng)的細(xì)胞相比顯著降低(p0.05)；術(shù)后6個月融合區(qū)鞏膜成纖維細(xì)胞力學(xué)環(huán)境培養(yǎng)后細(xì)胞的各項粘彈性參數(shù)(E0、E∞和μ)均增高(P0.05)；各組細(xì)胞3%拉伸組與6%拉伸組之間細(xì)胞的粘彈性參數(shù)(E0、E∞和μ)無明顯差異(P0.05)；三組細(xì)胞的粘彈性參數(shù)(E0、E∞和μ)在經(jīng)過力學(xué)拉伸后趨于相同。 實驗結(jié)果表明：力學(xué)刺激參與調(diào)節(jié)后鞏膜加固術(shù)后鞏膜成纖維細(xì)胞的生物學(xué)和生物力學(xué)行為,能夠提高正常鞏膜及后鞏膜加固術(shù)后原鞏膜組織的增殖活性,且能夠影響后鞏膜加固術(shù)后不同區(qū)域鞏膜成纖維細(xì)胞MMP-2、TIMP-2、TGF-β1及bFGF的表達(dá)及生物力學(xué)特性。力學(xué)刺激引起鞏膜成纖維細(xì)胞的這些變化有利于鞏膜胞外基質(zhì)的沉積、鞏膜成纖維細(xì)胞的增殖以及鞏膜組織與加固條帶之間融合,從而使鞏膜組織增厚,提高鞏膜的生物力學(xué)特性,控制近視的發(fā)展。
[Abstract]:Myopic myopia is a common eye disease , and has refractive myopia and progressive myopia . Among them , progressive myopia is also known as high myopia or pathological myopia , which is one of the most common causes of blindness . Posterior scleral reinforcement can improve the intensity of posterior sclera , prevent further prolongation of visual function , and prevent visual function from further worsening . It is a more effective method to treat high myopia .

In this paper , the mechanism of scleral reinforcement after posterior scleral reinforcement was studied in this paper . The mechanism of scleral reinforcement after scleral reinforcement after scleral reinforcement was studied . The mechanism of scleral reinforcement was explained from the perspective of mechanical - cell biology , which would provide theoretical guidance for the clinical selection and popularization of the operation .

The main research contents and conclusions are as follows :

1 . Forty - four New Zealand white rabbits who were not separated from breast - feeding were randomly and single - eye to establish a high myopia model , and the other one was the control eye . After induction , the eyes were opened , the dioptric power was measured and the length of the eye axis was measured . After induction , the relative myopia of 0.80 鹵 0.72D was induced in the experimental eye and the control eye . The difference was statistically significant ( P0.05 ) , indicating that the animal model was established successfully .

The expressions of MMP - 2 and TIMP - 2 , TGF - 尾1 and bFGF in scleral tissues were detected by immunohistochemistry . The results showed that the expression of MMP - 2 and TIMP - 2 , TGF - 尾1 and bFGF increased . These changes were beneficial to the proliferation of scleral fibroblasts , inhibited the degradation of scleral matrix protein , promoted the synthesis of extracellular matrix , and thickened scleral tissue .

3 . Tissue culture method was used to extract and culture the normal scleral tissue . After 3 months of operation , the scleral fibroblasts were cultured in the scleral tissue and the fusion area . The results showed that Vimentin was positive , Desmin staining was negative , keratin staining was negative , S - 100 was negative , and it was confirmed that the obtained cells were scleral fibroblasts .

ATP fluorescence assay was used to detect the proliferative activity of each group of cells .
The expressions of MMP - 2 , TIMP - 2 , TGF - 尾1 and bFGF were detected by ELISA .
The viscoelastic parameters ( instantaneous modulus E0 , equilibrium modulus E 鈭，

本文編號：1978315