腺病毒介导的色素上皮衍生因子抑制大鼠脉络膜新生血管的研究

An experimental study on choroidal neovascularization inhibited by adenoviral vectored pigment eplthelium-derived factor

目的 探讨腺病毒介导的色素上皮衍生因子（AdPEDF）对大鼠脉络膜新生血管（CNV）的抑制作用。方法 实验研究。选取6～8周雌性BN大鼠68只（136只眼），CNV模型建立后，采用单纯随机抽样方法将68只大鼠随机分为5组，空白对照组（N组）4只大鼠（8只眼），其余64只大鼠（128只眼）作为实验组。根据给药方式不同将实验组随机分为玻璃体腔注射实验组（A组）、玻璃体腔注射对照组（B组）、球周注射实验组（C组）、球周注射对照组（D组）。A组玻璃体腔注射AdPEDF1μl；B组玻璃体腔注射腺病毒载体注射液（AdNull）1μl；C组球周注射AdPEDF1山；D组球周注射AdNull1μl。于给药后3、7、14及28d行组织病理、TUNEL染色、荧光素眼底血管造影（FFA）检查、及CNV中央最大厚度测量。应用SPSS11．5统计学软件对光凝斑荧光素渗漏行秩和检验；对CNV发生率行X^2检验；对CNV中央最大厚度行单因素与析因方差分析。结果 A组（54．7％）和C组（56．3％）给药后比给药前荧光素渗漏减轻（t=2．75，3．15；P〈0．01）。给药后7d，A组（57．3％）、C组（57．8％）CNV数量减少；B、D组CNV呈显著纤维血管增殖。给药后A、C组CNV中央最大厚度分别为（44．51±0．53）和（44．37±0．48）μm，较N组减小（F：7．57，8．85；P〈0．01），并且随时间延长而减小（F=4．31，5．25；P〈0．05）。给药后3dA组CNV中央最大厚度为（46．35±0．62）μm，比C组（44．90±0．44）μm大（F=3．55，P〈0．05）；给药后14及28d，A组CNV中央最大厚度比C组减少（F=6．54，P〈0．01；F=4．41，P〈0．05）。给药后A、C组CNV内皮细胞出现部分TUNEL阳性细胞。术后并发症为白内障（5只眼）。结论 AdPEDF对BN大鼠CNV有抑制作用，治疗后7d起效，14d抑制作用最强，可持续至28d。玻璃体腔注射比球周注射起效慢，但抑制作用强。

Objective To study the effect of adenoviral vectored pigment epithelium-derived factor （AdPEDF） gene transfer on established neovascularization in a rat model of choroidal neovascularization （CNV）. Methods It was a experimental study. Sixty-eight female BN rats （136 eyes） with 6 to 8 weeks were used in this study. After the CNV model founded, 68 rats were divided into 5 groups with simple random sampling method. Four of them （8 eyes） were randomly selected as normal control group and 64 rats （ 128 eyes） were as experimental group. The experimental group included intravitreal injection with AdPEDF 1 μl （group A）, intravitreal injection with control vector （AdNull） 1 μl （group B）, periocular injection with AdPEDF 1μl （group C ）, and periocular injection with AdNull 1μl （group D）. Histopathology, TUNEL staining, fundus fluorescein angiography （FFA） , and thickness of CNV were tested 3, 7, 14, and 28 days after injection. Results （ 1 ） The leakages appeared decrease after treatment in group A （54. 7% ） andC （56.3%）（t =2.75,t =3.15;P 〈0.01）. （2） CNV decreased in group A （57.3%） and C （57.8%） 7 days after injection, and kept fibrovascular proliferation in group B and D. （3）The thickness of CNV in group A [（44.51±0.53） μm] and C [（44.37±0.48） μm] was significantly less than that in normal control group [ （ 46. 35± 0. 93 ） μm ] after the treatment （ F = 7. 57,8. 85 ; P 〈 0. 01 ） , and it diminished with the time prolong （ F = 4. 31,5.25 ; P 〈 0. 05 ）. The thickness of CNV in group A [ （46. 35 ±0. 62） μm ] was greater than that in group C [ （44.90 ± 0. 44 ） μm ] 3 days after treatment （ F = 3.55, P 〈 0. 05）, and it was less in group A than that in group C 14 and 28 days after treatment （F=6. 54,P 〈0. 01 ; F = 4. 41, P 〈 0. 05 ）. （4） There were positive TUNEL stainings in choroidal neovascular endothelium in group A and C. （5） Postoperative complication included cataract （5 eyes） after intravitreal injection. Conclusions AdPEDF is effective in inhibiting CNV in an animal model. The effect appears 7 days after treatment, reaches the peak on day 14, and keeps stable on day 28. The inhibition effect on CNV appears slowly in eyes with intravitreal injection than that with periocular injection, and it is stronger in eyes with intravitreal injection.