中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2011年
11期
988-992
,共5页
付浴东%周占宇%万金娥%丰慧%李松涛
付浴東%週佔宇%萬金娥%豐慧%李鬆濤
부욕동%주점우%만금아%봉혜%리송도
Avastin%视网膜/氧诱导视网膜新生血管化%血管形成%玻璃体腔注射
Avastin%視網膜/氧誘導視網膜新生血管化%血管形成%玻璃體腔註射
Avastin%시망막/양유도시망막신생혈관화%혈관형성%파리체강주사
Avastin%Retina/oxygen-induced retinal neovascularization%Angiogenesis%Intravitreal injection
背景 早产儿视网膜病变(ROP)主要源于视网膜新生血管的形成,avastin可以同血管内皮生长因子(VEGF)的所有异构体发生高亲和力结合,拮抗其生理活性.目的 观察玻璃体腔注射avastin对氧诱导视网膜病变模型新生血管的作用.方法 取7日龄清洁级C57BL/6J小鼠90只,用随机数字表法随机分为6组:空气对照组、高氧对照组、高氧BSS组和低、中、高剂量avastin组,每组15只.空气对照组小鼠在空气中喂养,其他各组均置于体积分数75％±2％O2的高氧环境中饲养,连续5d,高氧BSS组大鼠玻璃体腔注射无菌BSS液0.5μl,3个avastin处理组小鼠分别玻璃体腔注射1.25、2.50、5.00 g/L avastin各0.5μl.17日龄时过量麻醉处死小鼠,摘除眼球制备视网膜组织切片,苏木精—伊红染色后计数突破内界膜的新生血管内皮细胞核数目；应用免疫组织化学染色法检测视网膜中CD34分子的表达；利用视网膜铺片技术观察低、中、高剂量avastin组大鼠视网膜新生血管的形态并进行比较.结果苏木精—伊红染色发现,高氧对照组小鼠视网膜突破内界膜的新生血管内皮细胞核数目明显多于空气对照组,差异有统计学意义(P＜0.01)；不同剂量的avastin组小鼠视网膜中突破内界膜的新生血管内皮细胞核数明显少于高氧BSS组和高氧对照组,差异均有统计学意义(P＜0.01)；高剂量avastin组与低剂量avastin组比较,突破内界膜的新生血管内皮细胞核数明显减少,差异有统计学意义(P＜0.05).免疫组织化学检测显示,高氧对照组视网膜突破内界膜的内皮细胞CD34分子呈阳性表达.视网膜铺片可见空气对照组的血管自视盘向四周均匀放射,分支良好,结构清晰；高氧对照组、高氧BBS组可见大量新生血管,其结构紊乱,分布不均；各剂量avastin组随着剂量的增加,视网膜血管分布和走行接近空气对照组.结论玻璃体腔注射avastin可抑制视网膜新生血管的产生,其作用与avastin的剂量有关.
揹景 早產兒視網膜病變(ROP)主要源于視網膜新生血管的形成,avastin可以同血管內皮生長因子(VEGF)的所有異構體髮生高親和力結閤,拮抗其生理活性.目的 觀察玻璃體腔註射avastin對氧誘導視網膜病變模型新生血管的作用.方法 取7日齡清潔級C57BL/6J小鼠90隻,用隨機數字錶法隨機分為6組:空氣對照組、高氧對照組、高氧BSS組和低、中、高劑量avastin組,每組15隻.空氣對照組小鼠在空氣中餵養,其他各組均置于體積分數75％±2％O2的高氧環境中飼養,連續5d,高氧BSS組大鼠玻璃體腔註射無菌BSS液0.5μl,3箇avastin處理組小鼠分彆玻璃體腔註射1.25、2.50、5.00 g/L avastin各0.5μl.17日齡時過量痳醉處死小鼠,摘除眼毬製備視網膜組織切片,囌木精—伊紅染色後計數突破內界膜的新生血管內皮細胞覈數目；應用免疫組織化學染色法檢測視網膜中CD34分子的錶達；利用視網膜鋪片技術觀察低、中、高劑量avastin組大鼠視網膜新生血管的形態併進行比較.結果囌木精—伊紅染色髮現,高氧對照組小鼠視網膜突破內界膜的新生血管內皮細胞覈數目明顯多于空氣對照組,差異有統計學意義(P＜0.01)；不同劑量的avastin組小鼠視網膜中突破內界膜的新生血管內皮細胞覈數明顯少于高氧BSS組和高氧對照組,差異均有統計學意義(P＜0.01)；高劑量avastin組與低劑量avastin組比較,突破內界膜的新生血管內皮細胞覈數明顯減少,差異有統計學意義(P＜0.05).免疫組織化學檢測顯示,高氧對照組視網膜突破內界膜的內皮細胞CD34分子呈暘性錶達.視網膜鋪片可見空氣對照組的血管自視盤嚮四週均勻放射,分支良好,結構清晰；高氧對照組、高氧BBS組可見大量新生血管,其結構紊亂,分佈不均；各劑量avastin組隨著劑量的增加,視網膜血管分佈和走行接近空氣對照組.結論玻璃體腔註射avastin可抑製視網膜新生血管的產生,其作用與avastin的劑量有關.
배경 조산인시망막병변(ROP)주요원우시망막신생혈관적형성,avastin가이동혈관내피생장인자(VEGF)적소유이구체발생고친화력결합,길항기생리활성.목적 관찰파리체강주사avastin대양유도시망막병변모형신생혈관적작용.방법 취7일령청길급C57BL/6J소서90지,용수궤수자표법수궤분위6조:공기대조조、고양대조조、고양BSS조화저、중、고제량avastin조,매조15지.공기대조조소서재공기중위양,기타각조균치우체적분수75％±2％O2적고양배경중사양,련속5d,고양BSS조대서파리체강주사무균BSS액0.5μl,3개avastin처리조소서분별파리체강주사1.25、2.50、5.00 g/L avastin각0.5μl.17일령시과량마취처사소서,적제안구제비시망막조직절편,소목정—이홍염색후계수돌파내계막적신생혈관내피세포핵수목；응용면역조직화학염색법검측시망막중CD34분자적표체；이용시망막포편기술관찰저、중、고제량avastin조대서시망막신생혈관적형태병진행비교.결과소목정—이홍염색발현,고양대조조소서시망막돌파내계막적신생혈관내피세포핵수목명현다우공기대조조,차이유통계학의의(P＜0.01)；불동제량적avastin조소서시망막중돌파내계막적신생혈관내피세포핵수명현소우고양BSS조화고양대조조,차이균유통계학의의(P＜0.01)；고제량avastin조여저제량avastin조비교,돌파내계막적신생혈관내피세포핵수명현감소,차이유통계학의의(P＜0.05).면역조직화학검측현시,고양대조조시망막돌파내계막적내피세포CD34분자정양성표체.시망막포편가견공기대조조적혈관자시반향사주균균방사,분지량호,결구청석；고양대조조、고양BBS조가견대량신생혈관,기결구문란,분포불균；각제량avastin조수착제량적증가,시망막혈관분포화주행접근공기대조조.결론파리체강주사avastin가억제시망막신생혈관적산생,기작용여avastin적제량유관.
Background Retinopathy of prematurity is mainly due to retinal neovascularization.Objective This laboratory work was to evaluate the efficacy of different dosage of avastin for inhibiting retinal neovascularization.Methods Ninety 7-day-old clean C57BL/J6 mice were randomized into six groups as follows:air control group,hyperxia control group,hyperxia BSS group and avastin groups.C57BL/J6 mice in air control group were raised in regular air environments.The fifty mice were fed under the environment with 75％ ±2％ oxygen for 5 days to establish the retinal neovascularization models.The 1.25,2.50 and 5.00 g/L avastin (0.5 μl) were injected inteavtreally in forty-five mice models as low,moderate and high dosage avastin groups respectively,and 0.5 μl BSS was used at the same way in fifteen models as hyperxia BSS group.The mice were sacrificed in the 17-day-old age using excessive anesthesia method and the retina sections were prepared for the calculation of the numbers of vascular endothelial cell nuclei broken retinal inner membrane after hemotoxylin and eosin staining.The expression of CD34 in the retina was detected by immunochemistry.The morphology and distribution of retinal neovascular vessel in various groups were observed using retinal flat.The use of the animals followed the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The numbers of cell nuclei broken the inner limiting membrane was significant increased in the hyperxia group compared with the air control group( P＜0.01 ),and those in difference doses of avastin were considerably reduced in comparison with hyperxia BSS group (P＜0.01) and hyperxia group (P＜0.01 ).The decrease of numbers of cell nuclei broken the inner limiting membrane was obvious in low dose of high dose of avastin compared with low dose of avastin (P＜0.05 ).CD34 was positively expressed in retina internal membrane of hyperxia group.Retinal flat revealed the regular distribution and normal structure of retinal vessels in air control group and avastin groups.However,retinal and vitreous cavity neovascularization,leakage and enlarged non-perfusion regions in the perimeter of the retina were seen in hyperxia group and hyperxia BSS group. Conclusions Intravitreal injection of avastin can arrest retinal angiogenesis in oxygen-induced retinal neovascularization models in a dose-dependent manner.
