中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2011年
6期
557-560
,共4页
陈松%于华香%粱泽玉%李文博
陳鬆%于華香%粱澤玉%李文博
진송%우화향%량택옥%리문박
脉络膜新生血管化/病因学%基因表达%眼镜蛇毒液类%疾病模型%动物
脈絡膜新生血管化/病因學%基因錶達%眼鏡蛇毒液類%疾病模型%動物
맥락막신생혈관화/병인학%기인표체%안경사독액류%질병모형%동물
Choroidal neovascularization/etiology%Gene expression%Cobra venoms%Disease models,animal
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0.1 μg/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)和链霉亲和素生物素过氧化物酶复合物-荧光素异硫氰酸盐(SABC-FITC)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-Pro Plus 6.0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.
目的 觀察激光誘導小鼠脈絡膜新生血管(CNV)中C9的錶達及眼鏡蛇毒因子(CVF)對CNV的抑製作用.方法 C57BL/6J小鼠36隻隨機分為對照組、單純激光光凝組、CVF榦預組,每組12隻.後兩組用激光光凝方式建立CNV模型.CVF榦預組在激光光凝前2d和激光光凝後每天以0.1 μg/g的劑量腹腔註射CVF;激光光凝後6d,採用熒光素眼底血管造影檢查確定單純激光光凝組小鼠CNV形成.其後1、3、5、7d,斷頸處死各組小鼠後摘除眼毬作冰凍切片.採用囌木精-伊紅(HE)和鏈黴親和素生物素過氧化物酶複閤物-熒光素異硫氰痠鹽(SABC-FITC)免疫熒光組織化學染色法觀察膜攻擊複閤物(MAC)中C9的錶達.連續冠狀切片後,選取免疫熒光SABC-FITC染色切片組織像,採用Image-Pro Plus 6.0圖像處理分析軟件測量平均纍積吸光度[A,舊稱光密度(OD)]值.結果 單純激光光凝組CNV形成後7d,單純激光光凝組仍可見CNV,CVF榦預組未見CNV形成.單純激光光凝組CNV形成後1、3、5、7d,單純激光光凝組均可見C9錶達,CVF榦預組均未見C9錶達.對照組、單純激光光凝組及CVF榦預組3組間平均纍積A值比較,差異有統計學意義(F=146.045,P=0.000).各組組內在單純激光光凝組CNV形成後1、3、5、7d各時間點平均纍積A值比較,差異也有統計學意義(F=9.725,P=0.000).結論 激光誘導小鼠CNV中存在C9錶達；CVF誘導補體消耗可抑製CNV形成.
목적 관찰격광유도소서맥락막신생혈관(CNV)중C9적표체급안경사독인자(CVF)대CNV적억제작용.방법 C57BL/6J소서36지수궤분위대조조、단순격광광응조、CVF간예조,매조12지.후량조용격광광응방식건립CNV모형.CVF간예조재격광광응전2d화격광광응후매천이0.1 μg/g적제량복강주사CVF;격광광응후6d,채용형광소안저혈관조영검사학정단순격광광응조소서CNV형성.기후1、3、5、7d,단경처사각조소서후적제안구작빙동절편.채용소목정-이홍(HE)화련매친화소생물소과양화물매복합물-형광소이류청산염(SABC-FITC)면역형광조직화학염색법관찰막공격복합물(MAC)중C9적표체.련속관상절편후,선취면역형광SABC-FITC염색절편조직상,채용Image-Pro Plus 6.0도상처리분석연건측량평균루적흡광도[A,구칭광밀도(OD)]치.결과 단순격광광응조CNV형성후7d,단순격광광응조잉가견CNV,CVF간예조미견CNV형성.단순격광광응조CNV형성후1、3、5、7d,단순격광광응조균가견C9표체,CVF간예조균미견C9표체.대조조、단순격광광응조급CVF간예조3조간평균루적A치비교,차이유통계학의의(F=146.045,P=0.000).각조조내재단순격광광응조CNV형성후1、3、5、7d각시간점평균루적A치비교,차이야유통계학의의(F=9.725,P=0.000).결론 격광유도소서CNV중존재C9표체；CVF유도보체소모가억제CNV형성.
Objective To observe the expression of C9 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 (0.1 μg/g)2 days before and once a day after laser treatment.Six days after photocoagulation,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.
