本文關(guān)鍵詞： 超聲 微泡造影劑 脈絡(luò)膜新生血管 血管內(nèi)皮生長(zhǎng)因子 抗血管內(nèi)皮生長(zhǎng)因子單克隆抗體　出處：《重慶醫(yī)科大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】： 脈絡(luò)膜新生血管(CNV)與許多眼底疾病有關(guān),是致盲的常見原因[1]。而目前的治療方法存在許多缺點(diǎn),如:方法復(fù)雜、昂貴、療效欠穩(wěn)定等。由于在CNV形成和發(fā)展過(guò)程中血管內(nèi)皮生長(zhǎng)因子(VEGF)發(fā)揮著中心調(diào)控作用,阻斷VEGF的作用已成為目前國(guó)內(nèi)外CNV治療方案中研究的熱點(diǎn)和重點(diǎn)。因此近年有學(xué)者將被美國(guó)FDA批準(zhǔn)的通過(guò)抑制血管生成發(fā)揮抗癌作用的新藥Avastin(即bevacizumab,血管內(nèi)皮生長(zhǎng)因子單克隆抗體)用于眼部新生血管性疾病,特別是年齡相關(guān)性黃斑變性的治療,并取得了較好的治療效果。而超聲微泡造影劑作為一種新型聲學(xué)造影劑,爆破過(guò)程中可有效增加局部組織通透性,提高局部藥物濃度,以達(dá)到靶向治療作用。本研究首先利用氬綠激光構(gòu)建了兔眼脈絡(luò)膜新生血管模型,其次將超聲爆破微泡與抗血管內(nèi)皮生長(zhǎng)因子單克隆抗體Avastin (bevacizumab)聯(lián)合,用于兔CNV的治療,觀察VEGF蛋白的表達(dá)和對(duì)CNV的抑制情況,為臨床治療CNV提供了一種新的思路。 方法:動(dòng)物模型的建造:用氬綠激光(波長(zhǎng)514.5nm)距視乳頭2~3個(gè)視盤直徑的顳側(cè)髓線上下視盤網(wǎng)膜密集處照射20個(gè)點(diǎn),激光光斑間隔300μm,激光光斑直徑50μm,激光功率0.7 W,曝光時(shí)間0.1 s,建造實(shí)驗(yàn)性色素兔脈絡(luò)膜新生血管模型。在造模后第21d進(jìn)行組織病理學(xué)和FFA觀察,確定CNV的形成。 造模成功后再進(jìn)行以下研究:將CNV兔分為對(duì)照組:不做任何處理; Avastin組(A):玻璃體內(nèi)注射Avastin;超聲微泡+Avastin組(U+MB+A):玻璃體腔內(nèi)注射Avastin及超聲微泡,再用頻率1MHZ,聲強(qiáng)分別為0.5W/cm2超聲輻照眼球60s,工作時(shí)間20%超,每周兩次,處理后7d、14d、28d用免疫熒光及Western-blot檢測(cè)VEGF蛋白表達(dá),FFA觀察CNV的抑制情況。 結(jié)果:在分組處理后7d、14d、28d超聲微泡+Avastin組免疫熒光及Western-blot檢測(cè)VEGF蛋白表達(dá)均明顯低于Avastin組(p0.05),FFA結(jié)果顯示超聲微泡+Avastin組熒光滲漏吸光度(A值)明顯低于Avastin組(p0.05),且與對(duì)照組差異均有統(tǒng)計(jì)學(xué)意義(p0.05)。以28天作為療效判定時(shí)間點(diǎn),FFA檢查結(jié)果顯示Avastin組熒光滲漏平均強(qiáng)度為66.96±4.41 ,與對(duì)照組(119.60±6.57 )相比,差異有統(tǒng)計(jì)學(xué)意義(t=16.2952;p=0.0000);超聲微泡+Avastin組為54.75±4.41,與Avastin組相比,差異有統(tǒng)計(jì)學(xué)意義(t=4.7955;p=0.0000)。且各組熒光滲漏強(qiáng)度隨時(shí)間變化均呈下降趨勢(shì)。分組處理后28d,組織免疫熒光及Western-blot檢測(cè)VEGF蛋白表達(dá)結(jié)果顯示,Avastin組為23.9825±3.3180與0.5666±0.0179,與對(duì)照組相比,差異有統(tǒng)計(jì)學(xué)意義(t=7.0327,p=0.0000;t=9.2596,P=0.0000);超聲微泡+Avastin組為19.5636±1.5006與0.3214±0.030 ,與Avastin組相比,差異有統(tǒng)計(jì)學(xué)意義(t=2.9724,p=0.0140;t=17.1 937,p=0.0000),且各組VEGF蛋白表達(dá)隨時(shí)間變化呈下降趨勢(shì)。 結(jié)論: 1、采用一定波長(zhǎng)、功率的氬綠激光進(jìn)行視網(wǎng)膜光凝方法,可較好復(fù)制實(shí)驗(yàn)性色素兔脈絡(luò)膜新生血管模型。 2、玻璃體腔注射Avastin,對(duì)脈絡(luò)膜新生血管有一定療效 3、超聲微泡與Avastin聯(lián)合使用時(shí),可有效增加Avastin對(duì)脈絡(luò)膜新生血管的治療效果。
[Abstract]:Choroidal neovascularization (CNV) associated with many fundus diseases, [1]. is a common cause of blindness and the current treatment methods have many disadvantages, such as complex, expensive, less effective and stable. The formation and development of vascular endothelial growth factor in the process of CNV (VEGF) plays a key role in regulating, blocking VEGF the effect has become the hotspot and focus of research at home and abroad CNV therapy. So in recent years scholars will be approved by the FDA to play a role in cancer drug Avastin through inhibition of angiogenesis (bevacizumab, vascular endothelial growth factor monoclonal antibody) for ocular neovascular diseases, especially in the treatment of age-related macular degeneration. And get a better therapeutic effect. Ultrasound microbubble contrast agent as a new contrast agent, the blasting process can effectively increase the local tissue permeability, improve local drug In order to achieve the concentration, the effect of targeted therapy. In this study, the use of argon green laser was constructed rabbit choroidal neovascularization model, followed by ultrasonic microbubble and anti VEGF monoclonal antibody Avastin (bevacizumab), for the treatment of rabbit CNV, to observe the expression of VEGF protein and inhibition of CNV. Provides a new way for the treatment of CNV.
Methods: to build animal model with argon green laser (wavelength 514.5nm) from the optic disc diameter 2~3 temporal pith line of optic disc intensive irradiation omentum 20 points, laser spot spacing 300 m, laser spot diameter of 50 m and 0.7 W laser power, exposure time of 0.1 s, the construction of experimental pigmented choroidal neovascularization in rabbits model. After modeling the 21d pathology and FFA observation, determine the formation of CNV.
The following research again after the success of the model: the CNV rabbits were divided into control group: no treatment; group Avastin (A): Intravitreal injection of Avastin; ultrasound microbubble group +Avastin (U+MB+A): Intravitreal injection of Avastin and ultrasound, then the frequency 1MHZ, sound intensity were 0.5W/cm2 ultrasound irradiation of eyeball 60s, working time of 20%, two times a week, after the treatment of 7D, 14d, 28d, VEGF protein was detected by immunofluorescence and the expression of Western-blot, inhibition of FFA observation of CNV.
Results: in group 7d after treatment, 14d, 28d ultrasound microbubble group +Avastin immunofluorescence detection of VEGF protein and Western-blot expression were significantly lower than that of Avastin group (P0.05), FFA results showed that ultrasound microbubble group +Avastin fluorescence leakage absorbance (A value) was significantly lower than that of Avastin group (P0.05) and the control group, and the differences were statistically significant (P0.05) for 28 days. As the effect of the judgment time, FFA examination showed that Avastin group average fluorescence leakage intensity was 66.96 + 4.41, and the control group (119.60 + 6.57) compared to the difference was statistically significant (t=16.2952; p=0.0000); ultrasoundmicrobubble +Avastin group is 54.75 + 4.41, compared with the Avastin group, a statistically significant differences (t=4.7955; p=0.0000). And each group of fluorescence leakage intensity decreased. After 28d, the expression of VEGF by immunofluorescence and the expression of Western-blot showed that the Avastin group is 23.9825 + 3.3180 With 0.5666 + 0.0179, compared with the control group, the difference was statistically significant (t=7.0327, p=0.0000; t=9.2596, P=0.0000); ultrasoundmicrobubble +Avastin group is 19.5636 + 1.5006 and 0.3214 + 0.030, compared with the Avastin group, the difference was statistically significant (t=2.9724, p=0.0140; t=17.1 937, p=0.0000), and each group of VEGF expression downward trend changes with time.
Conclusion:
1, the retinal photocoagulation method with a certain wavelength and power argon green laser can be used to reproduce the experimental pigmented rabbit choroidal neovascularization model.
2, the intravitreal injection of Avastin has a certain effect on the choroidal neovascularization.
3, the combined use of ultrasound microbubbles and Avastin can effectively increase the therapeutic effect of Avastin on choroidal neovascularization.

【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R774.1

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