目的 研究高氧诱导小鼠视网膜病变模型中血管内皮生长因子(VEGF)和色素上皮衍生因子(PEDF)的表达和意义,并探讨两者在视网膜新生血管形成过程中的作用.方法 对照实验研究.对新生C57BL-6N系小鼠给予高氧后,置相对低氧环境中饲养,诱导产生视网膜新生血管.在小鼠生后第12、14及17天摘除眼球,通过逆转录聚合酶链反应和免疫印迹法及荧光素灌注造影,分别检测不同时间点全视网膜新生血管组织对VEGF与PEDF的mRNA和蛋白质的表达水平的差异,并观察视网膜新生血管的分布与形态变化,通过血管内皮细胞计数对新生血管进行量化.应用SPSS11.5统计学软件,采用析因设计方差分析,分别比较实验组与对照组mRNA与蛋白质表达水平的差异,以P<0.05作为差异有统计学意义.结果 在高氧环境中(出生第12天小鼠),OIR模型鼠VEGF蛋白质表达A值(0.47±0.12)较正常鼠(1.81±050)下降,PEDF蛋白质表达A值(5.35±0.94)较正常鼠(0.68±0.17)明显升高;而相对低氧环境中(出生第14和17天小鼠),VEGF蛋白质表达A值(2.15±0.46,5.49±0.97)较正常鼠(0.90±0.05,0.88±0.91)明显升高,PEDF蛋白质表达A值(2.07±0.35,1.37±0.48)较正常鼠(2.62±0.68,5.30±0.59)明显下降,尤以第17天下降显著.对不同时间点VEGF和PEDF蛋白质表达水平进行析因方差分析,结果显示各时间点的VEGF蛋白质表达水平差异有统计学意义(F=70.450,P=0.000),各时间点的PEDF蛋白质表达水平差异有统计学意义(F=160.237,P=0.000).出生第12、14及17天小鼠视网膜VEGF蛋白质的表达与正常对照组比较,差异有统计学意义(P=0.009,0.010,0.000),PEDF蛋白质的表达差异也有统计学意义(P=0.002,0.046,0.000);出生第12、14及17天小鼠视网膜VEGF mRNA的表达与正常对照组比较,差异有统计学意义(P=0.001,0.000,0.001),PEDF mRNA的表达差异也有统计学意义(P=0.000,0.001,0.000).伴随着视网膜组织的缺氧,出现了VEGF蛋白质表达水平升高和PEDF蛋白质表达水平降低,导致视网膜组织中VEGF与PEDF蛋白质表达失衡;VEGF与PEDF的mRNA表达亦发生类似变化,且较蛋白质改变提前发生.上述改变均伴随着同期视网膜新生血管的发生、发展及其严重程度而逐渐加重.结论 VEGF和PEDF的mRNA和蛋白质表达失衡可能足造成视网膜新生血管发生的机制之一.(中华眼科杂志,2008,44:734-740)
目的 研究高氧誘導小鼠視網膜病變模型中血管內皮生長因子(VEGF)和色素上皮衍生因子(PEDF)的錶達和意義,併探討兩者在視網膜新生血管形成過程中的作用.方法 對照實驗研究.對新生C57BL-6N繫小鼠給予高氧後,置相對低氧環境中飼養,誘導產生視網膜新生血管.在小鼠生後第12、14及17天摘除眼毬,通過逆轉錄聚閤酶鏈反應和免疫印跡法及熒光素灌註造影,分彆檢測不同時間點全視網膜新生血管組織對VEGF與PEDF的mRNA和蛋白質的錶達水平的差異,併觀察視網膜新生血管的分佈與形態變化,通過血管內皮細胞計數對新生血管進行量化.應用SPSS11.5統計學軟件,採用析因設計方差分析,分彆比較實驗組與對照組mRNA與蛋白質錶達水平的差異,以P<0.05作為差異有統計學意義.結果 在高氧環境中(齣生第12天小鼠),OIR模型鼠VEGF蛋白質錶達A值(0.47±0.12)較正常鼠(1.81±050)下降,PEDF蛋白質錶達A值(5.35±0.94)較正常鼠(0.68±0.17)明顯升高;而相對低氧環境中(齣生第14和17天小鼠),VEGF蛋白質錶達A值(2.15±0.46,5.49±0.97)較正常鼠(0.90±0.05,0.88±0.91)明顯升高,PEDF蛋白質錶達A值(2.07±0.35,1.37±0.48)較正常鼠(2.62±0.68,5.30±0.59)明顯下降,尤以第17天下降顯著.對不同時間點VEGF和PEDF蛋白質錶達水平進行析因方差分析,結果顯示各時間點的VEGF蛋白質錶達水平差異有統計學意義(F=70.450,P=0.000),各時間點的PEDF蛋白質錶達水平差異有統計學意義(F=160.237,P=0.000).齣生第12、14及17天小鼠視網膜VEGF蛋白質的錶達與正常對照組比較,差異有統計學意義(P=0.009,0.010,0.000),PEDF蛋白質的錶達差異也有統計學意義(P=0.002,0.046,0.000);齣生第12、14及17天小鼠視網膜VEGF mRNA的錶達與正常對照組比較,差異有統計學意義(P=0.001,0.000,0.001),PEDF mRNA的錶達差異也有統計學意義(P=0.000,0.001,0.000).伴隨著視網膜組織的缺氧,齣現瞭VEGF蛋白質錶達水平升高和PEDF蛋白質錶達水平降低,導緻視網膜組織中VEGF與PEDF蛋白質錶達失衡;VEGF與PEDF的mRNA錶達亦髮生類似變化,且較蛋白質改變提前髮生.上述改變均伴隨著同期視網膜新生血管的髮生、髮展及其嚴重程度而逐漸加重.結論 VEGF和PEDF的mRNA和蛋白質錶達失衡可能足造成視網膜新生血管髮生的機製之一.(中華眼科雜誌,2008,44:734-740)
목적 연구고양유도소서시망막병변모형중혈관내피생장인자(VEGF)화색소상피연생인자(PEDF)적표체화의의,병탐토량자재시망막신생혈관형성과정중적작용.방법 대조실험연구.대신생C57BL-6N계소서급여고양후,치상대저양배경중사양,유도산생시망막신생혈관.재소서생후제12、14급17천적제안구,통과역전록취합매련반응화면역인적법급형광소관주조영,분별검측불동시간점전시망막신생혈관조직대VEGF여PEDF적mRNA화단백질적표체수평적차이,병관찰시망막신생혈관적분포여형태변화,통과혈관내피세포계수대신생혈관진행양화.응용SPSS11.5통계학연건,채용석인설계방차분석,분별비교실험조여대조조mRNA여단백질표체수평적차이,이P<0.05작위차이유통계학의의.결과 재고양배경중(출생제12천소서),OIR모형서VEGF단백질표체A치(0.47±0.12)교정상서(1.81±050)하강,PEDF단백질표체A치(5.35±0.94)교정상서(0.68±0.17)명현승고;이상대저양배경중(출생제14화17천소서),VEGF단백질표체A치(2.15±0.46,5.49±0.97)교정상서(0.90±0.05,0.88±0.91)명현승고,PEDF단백질표체A치(2.07±0.35,1.37±0.48)교정상서(2.62±0.68,5.30±0.59)명현하강,우이제17천하강현저.대불동시간점VEGF화PEDF단백질표체수평진행석인방차분석,결과현시각시간점적VEGF단백질표체수평차이유통계학의의(F=70.450,P=0.000),각시간점적PEDF단백질표체수평차이유통계학의의(F=160.237,P=0.000).출생제12、14급17천소서시망막VEGF단백질적표체여정상대조조비교,차이유통계학의의(P=0.009,0.010,0.000),PEDF단백질적표체차이야유통계학의의(P=0.002,0.046,0.000);출생제12、14급17천소서시망막VEGF mRNA적표체여정상대조조비교,차이유통계학의의(P=0.001,0.000,0.001),PEDF mRNA적표체차이야유통계학의의(P=0.000,0.001,0.000).반수착시망막조직적결양,출현료VEGF단백질표체수평승고화PEDF단백질표체수평강저,도치시망막조직중VEGF여PEDF단백질표체실형;VEGF여PEDF적mRNA표체역발생유사변화,차교단백질개변제전발생.상술개변균반수착동기시망막신생혈관적발생、발전급기엄중정도이축점가중.결론 VEGF화PEDF적mRNA화단백질표체실형가능족조성시망막신생혈관발생적궤제지일.(중화안과잡지,2008,44:734-740)
Objective To investigate the expression and significance of vascular endothelial growth factor (VEGF) and pigment epithelium derived factor (PEDF) in oxygen-induced mouse retinopathy. Methods This experiment was a control experiment study. Newborn C57BL-6 mice were exposed to hyperoxia, and then returned to normoxia to induce retinal neovascularization. Mice were sacrificed at postnatal days 12, 14 and 17 and the retina were processed for RT-PCR, Western Blot and fluorescein angiography. Analysis of variance and Dunnett's t3 was used to compare the mRNA and protein level of VEGF and PEDF between experiment and control groups. Statistical difference was considered significant at a P value less than 0. 05. Results At any of the time points tested, there was significant difference at VEGF protein level (A value) between the experiment (0.47±0.12,2.15±0.46,5.49±0.97) and control(1.81±0. 50,0.90±0.05,0.88±0.91) groups(P=0. 009,0. 010,0. 000, respectively);the same situation occurred at PEDF protein level (P=0. 002,0. 046,0. 000,respectively) . At postnatal day 12,14 and 17 ,a significant difference at VEGF protein level was observed among the different experiment groups (P =0. 002,0. 001,0. 000, respectively); the same situation occurred at PEDF protein level( P =0.009,0.010,0.000, respectively). There was significant difference at VEGF mRNA level between the experiment and control groups (P= 0. 001,0. 000,0. 001);at postnatal day 12 and 14, the same situation occurred at PEDF protein level(P=0.001,0.000, respectively) ,but there was no significant difference at postnatal day 17(P=0.612). At postnatal day 12,14 and 17,a significant difference at VEGF mRNA level was observed among different experiment groups (P=0.000, 0.001,0.000, respectively); the same situation occurred at PEDF mRNA level (P=0.000,0.001,0.000, respectively). The time course of the decrease of PEDF was consistent with the increase of VEGF expressiorL VEGF/PEDF ratio change was correlated with the development and progression of retinal neovascularization. The time course of PEDF mRNA down-regulation was consistent with the VEGF mRNA up-regulation, similar with the changes of the protein. The change of VEGF and PEDF mRNA was prior to that of the protein. Conclusions One of the mechanisms for development of retinal neovascularization is the changes of VEGF\PEDF level in the retina.(Chin J Ophthalmol, 2008,44:734-740)
