激光诱导小鼠脉络膜新生血管中C9的表达及眼镜蛇毒因子的抑制作用

C9 expression in the laser-induced choroidai neovascularization and cobra venom factor inhibits choroidal neovascularization in mice

目的观察激光诱导小鼠脉络膜新生血管（CNV）中C9的表达及眼镜蛇毒因子（CVF）对CNV的抑制作用。方法C57BL／6J小鼠36只随机分为对照组、单纯激光光凝组、CVF干预组，每组12只。后两组用激光光凝方式建立CNV模型。CVF干预组在激光光凝前2d和激光光凝后每天以0．1μg／g的剂量腹腔注射CVF；激光光凝后6d，采用荧光素眼底血管造影检查确定单纯激光光凝组小鼠CNV形成。其后1、3、5、7d，断颈处死各组小鼠后摘除眼球作冰冻切片。采用苏木精-伊红（HE）和链霉亲和素生物索过氧化物酶复合物一荧光素异硫氰酸盐（SABDFITc）免疫荧光组织化学染色法观察膜攻击复合物（MAC）中c9的表达。连续冠状切片后，选取免疫荧光SABC-FITC染色切片组织像，采用Image—ProPlus6．0图像处理分析软件测量平均累积吸光度[A，旧称光密度（OD）]值。结果单纯激光光凝组CNV形成后7d，单纯激光光凝组仍可见CNV，CVF干预组未见CNV形成。单纯激光光凝组CNV形成后1、3、5、7d，单纯激光光凝组均可见c9表达，CVF干预组均未见C9表达。对照组、单纯激光光凝组及CVF干预组3组间平均累积A值比较，差异有统计学意义（F=146．045，P=0．000）。各组组内在单纯激光光凝组CNV形成后1、3、5、7d各时间点平均累积A值比较，差异也有统计学意义（F=9．725，P=0．000）。结论激光诱导小鼠CNV中存在C9表达；CVF诱导补体消耗可抑制CNV形成。

Objective To observe the expression of CO in laser-induced choroidal;neovascularization （CNV） and the inhibitory effect of cobra venom factor （CVF） on CNV. Methods Thirty-six C57BL/6J mice were randomly divided into control group （n= 12）, laser photocoagulation group （n= 12） and CVF group （n=12）. The mice in control group had no interference treatment. CNV of the latter 2 groups were induced by photocoagulation, The mice of CVF group received intraperitoneal injection of CVF （0.1μg/g） 2 days before and once a day after laser treatment. Six days after pbotocoagulation, the examination of fundus fluorescein angiography was used to confirm that CNV existed in the mice of laser the photocoagulation group. Then at 1, 3, 5, 7 days, the C9 expression was observed using SABC-FITC histochemical stain and HE stain. The absorbance was measured by Image-Pro Plus 6.0 image software. Results Only in the laser group had formed CNV. At different time points, C9 expression was observed in the laser photocoagulation group but not in the CVF group. The A value showed a significant difference among 3 groups at different time points （F= 146. 045, P=0. 000）. The significant difference of A value exited in each group at different time points （F= 9. 725, P= 0. 000）. Conclusions There is C9 expression in laser-induced choroidal neovascularization. CVF can inhibit CNV formation by inducing complement consumption.