本文選題：角膜新生血管 + 堿燒傷�。� 參考：《中國(guó)人民解放軍醫(yī)學(xué)院》2013年博士論文

【摘要】：目的:檢測(cè)靶向Bcl-2小分子化合物納米懸混液（Z24-NS）在眼部應(yīng)用安全性基礎(chǔ)上，觀察Z24-NS對(duì)大鼠角膜堿燒傷后角膜新生血管（cornealneovascularization，CNV）增殖的影響，并初步探討其抑制效應(yīng)及其可能的機(jī)制，為將來Z24-NS滴眼液應(yīng)用于臨床奠定理論基礎(chǔ)。 方法： 1．8只SD大鼠隨機(jī)分為Z24組和生理鹽水組兩組，，每組4只(4眼)。Z24組左眼滴2％的Z24-NS，生理鹽水組左眼滴0.9％的生理鹽水。連續(xù)滴眼14天，通過行為學(xué)觀察、裂隙燈顯微鏡檢查、制作病理切片，觀察光鏡下角膜病理學(xué)改變，檢測(cè)2％Z24-NS滴眼液有無明顯毒副作用。 2．36只SD大鼠，采用浸潤(rùn)1mol/L NaOH的3×3mm濾紙貼附左眼角膜中央40s的方法制作大鼠角膜堿燒傷誘導(dǎo)CNV模型，并隨機(jī)分為Z24-NS滴眼液治療組、地塞米松磷酸鈉滴眼液陽性對(duì)照組以及空白組三組，每組12只(12眼)。Z24組左眼滴2％的Z24-NS，地米組左眼滴0.1％地塞米松磷酸鈉滴眼液，每日4次，空白組不做任何處理。實(shí)驗(yàn)周期為14天。在堿燒傷后3天、7天、14天通過裂隙燈顯微鏡檢查及照相定性、定量觀察各組的CNV生長(zhǎng)情況并計(jì)算CNV面積、角膜混濁程度、炎癥反應(yīng)和前房反應(yīng)。 3．堿燒傷后14天在第二部分的基礎(chǔ)上通過病理組織學(xué)觀察及計(jì)算機(jī)圖像分析系統(tǒng)分析各組角膜情況，檢測(cè)各組角膜組織中炎癥細(xì)胞的數(shù)量、微血管密度，通過免疫組織化學(xué)方法、RT-PCR和Western Blot檢測(cè)大鼠角膜VEGF及其受體Flk-1和Bcl-2的mRNA和蛋白的表達(dá)情況。 結(jié)果: 1．短期局部應(yīng)用2％的Z24-NS滴眼后大鼠無明顯甩頭、撓抓眼部現(xiàn)象，大鼠無角膜混濁、水腫，角膜上皮無明顯缺損，組織學(xué)檢查顯示結(jié)構(gòu)正常，未見炎癥細(xì)胞浸潤(rùn)。 2．堿燒傷后3、7、14天Z24組和地塞米松組CNV面積、炎癥指數(shù)及角膜混濁程度均小于空白組（P0.01或P0.05）。與地塞米松組比較，2％的Z24-NS抑制CNV程度、炎癥指數(shù)及角膜混濁程度差異無統(tǒng)計(jì)學(xué)意義(P0.05)。 3．堿燒傷后14天Z24組和地塞米松組角膜組織中炎癥細(xì)胞的數(shù)量、微血管密度均小于空白組（P0.01或P0.05），VEGF、 Flk-1及Bcl-2蛋白及mRNA表達(dá)明顯低于空白組（P0.05），而2％的Z24-NS與地塞米松組比較差異無統(tǒng)計(jì)學(xué)意義(P0.05)。各實(shí)驗(yàn)組VEGF和Flk-1的表達(dá)與MVD的數(shù)量呈正相關(guān)(r1=0.9997，P1=0.015；r2=0.9999，P2=0.011)。 結(jié)論： 1．2％的Z24-NS滴眼液短期局部應(yīng)用對(duì)大鼠的角膜結(jié)構(gòu)無影響，在眼部應(yīng)用是安全的。 2．2％的Z24-NS滴眼液對(duì)大鼠角膜堿燒傷后CNV的生長(zhǎng)具有明顯抑制作用，同時(shí)相應(yīng)減輕了角膜混濁程度，其抑制CRNV效果與0.1％地塞米松滴眼液相同。 3．2％的Z24-NS滴眼液有一定的抗炎作用，它可以抑制大鼠角膜新生血管模型中角膜VEGF、Flk-1及Bcl-2的表達(dá)，提示Z24-NS可能通過降低角膜VEGF、Flk-1及Bcl-2的產(chǎn)生，從而抑制了CNV的生長(zhǎng)。
[Abstract]:Objective : To investigate the effect of Z24 - NS on the proliferation of corneal neovascularization ( CNV ) after corneal alkali burn in rats and to investigate the inhibitory effect and possible mechanism of Z24 - NS on corneal neovascularization after corneal alkali burn in rats .

Method :

1.8 SD rats were randomly divided into two groups : Z24 - NS group and normal saline group .

2 . Thirty - six SD rats were randomly divided into two groups : Z24 - NS eye drops 2 % Z24 - NS , dexamethasone sodium phosphate eye drops 0.1 % sodium phosphate eye drops , 4 times a day .

3 . The expression of VEGF and its receptor Flk - 1 and Bcl - 2 mRNA and protein in corneal tissue were measured by pathological and histological observation and computer image analysis on the second part 14 days after alkali burn . The expression of VEGF and its receptor Flk - 1 and Bcl - 2 mRNA and protein were detected by immunohistochemistry , RT - PCR and Western Blot .

Results :

1 . After short - term topical application of 2 % Z24 - NS , no obvious spin - off was found in the rats , scratching the eyes , the rats had no corneal opacity , edema , no obvious defect of corneal epithelium , histological examination showed normal structure , and no infiltration of inflammatory cells was seen .

Compared with the dexamethasone group , 2 % of Z24 - NS inhibited the degree of CNV , the inflammatory index and the degree of corneal opacity ( P0.05 ) .

The expression of VEGF , Flk - 1 and Bcl - 2 protein and mRNA in the experimental group were significantly lower than those in the blank group ( P0.05 ) . The expression of VEGF and Flk - 1 was positively correlated with MVD ( r1 = 0.9997 , P1 = 0.015 ) .
r2=0.9999,P2=0.011).


Conclusion :

Short - term topical application of 1.2 % Z24 - NS eye drops has no effect on the corneal structure of rats and is safe in eye application .

2 . 2 % Z24 - NS eye drops significantly inhibited the growth of CNV after corneal alkali burn in rats , and the degree of corneal opacity was decreased accordingly , which inhibited the effects of CRNV and 0.1 % dexamethasone eye drops .

3.2 % Z24 - NS eye drops have anti - inflammatory effects , which can inhibit the expression of VEGF , Flk - 1 and Bcl - 2 in corneal neovascularization model in rats , suggesting that Z24 - NS may inhibit the growth of CNV by lowering the production of corneal VEGF , Flk - 1 and Bcl - 2 .

【學(xué)位授予單位】：中國(guó)人民解放軍醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類號(hào)】：R772.2

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本文編號(hào)：1801094