中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2008年
3期
200-206
,共7页
石璇%Irina Semkova%Norbert Kociok%Claudia Gavranic%Martina Becker%Antonia M.Joussen%Bernd Kirchhof
石璇%Irina Semkova%Norbert Kociok%Claudia Gavranic%Martina Becker%Antonia M.Joussen%Bernd Kirchhof
석선%Irina Semkova%Norbert Kociok%Claudia Gavranic%Martina Becker%Antonia M.Joussen%Bernd Kirchhof
脉络膜新生血管化%肿瘤坏死因子α%免疫球蛋白G%受体,肿瘤坏死因子%抗体,单克隆%光刺激
脈絡膜新生血管化%腫瘤壞死因子α%免疫毬蛋白G%受體,腫瘤壞死因子%抗體,單剋隆%光刺激
맥락막신생혈관화%종류배사인자α%면역구단백G%수체,종류배사인자%항체,단극륭%광자격
Choroidal neovascularization%Tumor necrosis factor-alpha%Immunoglobulin G%Recetor,tumor neicrosis factor%Antibodies,monoclonal%Photic stimulation
目的 探讨肿瘤坏死因子α(TNF-α)在激光诱导的小鼠脉络膜新生血管(CNV)形成中的作用.方法 野生型C57BL/6J小鼠36只(72只眼),每只眼视网膜上做4个光凝斑诱导CNV生成.小鼠分别于光凝前3 d或后3 d腹腔内植入渗透泵,给予TNF-α抑制剂依那昔普(5μg/h,n=12)或英利西单抗(5μg/h,n=12),持续7 d,对照组给予PBS(n=12).光凝前3 d的用药组分别于光凝后1和2周,光凝后3 d的用药组于光凝后10 d做荧光素眼底血管造影(FFA),之后立刻取左眼行视网膜组织学检查,右眼制作脉络膜平片,免疫组化方法 标记内皮细胞,测定荧光素渗漏面积及脉络膜平片CNV相关荧光面积,比较CNV病变的大小及程度.用免疫印迹法分析比较小鼠光凝前后视网膜色素上皮层及脉络膜上TNF-α蛋白的表达.结果 光凝后1周,免疫印迹法结果 显示小鼠脉络膜及视网膜色素上皮层TNF-α表达增加.光凝前3 d用药,光凝后1周,荧光素渗漏面积为(0.74±0.33)×104 像素,2周时则为(0.63±0.20)×104 像素,与对照组[(2.61±0.80)×104 像素]比较均显著减少(t=3.69,3.56;P＜0.05).光凝后1周对照组、依那昔普组和英利西单抗组CNV面积分别为(3.61±0.93)×105、(1.89±0.62)×105和(2.14±0.72)×105像素,与对照组比较,差异均有统计学意义(t=3.10,2.51;P＜0.05.但光凝后3 d用药,则只有依那昔普能显著抑制CNV病变.组织病理学检查进一步证实,治疗组小鼠的CNV病变直径及厚度小于对照组.结论 TNF-α参与激光诱导的CNV的形成.TNF-α抑制剂能抑制激光诱导的CNV的形成及渗漏.(中华眼科杂志,2008,44:200-206)
目的 探討腫瘤壞死因子α(TNF-α)在激光誘導的小鼠脈絡膜新生血管(CNV)形成中的作用.方法 野生型C57BL/6J小鼠36隻(72隻眼),每隻眼視網膜上做4箇光凝斑誘導CNV生成.小鼠分彆于光凝前3 d或後3 d腹腔內植入滲透泵,給予TNF-α抑製劑依那昔普(5μg/h,n=12)或英利西單抗(5μg/h,n=12),持續7 d,對照組給予PBS(n=12).光凝前3 d的用藥組分彆于光凝後1和2週,光凝後3 d的用藥組于光凝後10 d做熒光素眼底血管造影(FFA),之後立刻取左眼行視網膜組織學檢查,右眼製作脈絡膜平片,免疫組化方法 標記內皮細胞,測定熒光素滲漏麵積及脈絡膜平片CNV相關熒光麵積,比較CNV病變的大小及程度.用免疫印跡法分析比較小鼠光凝前後視網膜色素上皮層及脈絡膜上TNF-α蛋白的錶達.結果 光凝後1週,免疫印跡法結果 顯示小鼠脈絡膜及視網膜色素上皮層TNF-α錶達增加.光凝前3 d用藥,光凝後1週,熒光素滲漏麵積為(0.74±0.33)×104 像素,2週時則為(0.63±0.20)×104 像素,與對照組[(2.61±0.80)×104 像素]比較均顯著減少(t=3.69,3.56;P＜0.05).光凝後1週對照組、依那昔普組和英利西單抗組CNV麵積分彆為(3.61±0.93)×105、(1.89±0.62)×105和(2.14±0.72)×105像素,與對照組比較,差異均有統計學意義(t=3.10,2.51;P＜0.05.但光凝後3 d用藥,則隻有依那昔普能顯著抑製CNV病變.組織病理學檢查進一步證實,治療組小鼠的CNV病變直徑及厚度小于對照組.結論 TNF-α參與激光誘導的CNV的形成.TNF-α抑製劑能抑製激光誘導的CNV的形成及滲漏.(中華眼科雜誌,2008,44:200-206)
목적 탐토종류배사인자α(TNF-α)재격광유도적소서맥락막신생혈관(CNV)형성중적작용.방법 야생형C57BL/6J소서36지(72지안),매지안시망막상주4개광응반유도CNV생성.소서분별우광응전3 d혹후3 d복강내식입삼투빙,급여TNF-α억제제의나석보(5μg/h,n=12)혹영리서단항(5μg/h,n=12),지속7 d,대조조급여PBS(n=12).광응전3 d적용약조분별우광응후1화2주,광응후3 d적용약조우광응후10 d주형광소안저혈관조영(FFA),지후립각취좌안행시망막조직학검사,우안제작맥락막평편,면역조화방법 표기내피세포,측정형광소삼루면적급맥락막평편CNV상관형광면적,비교CNV병변적대소급정도.용면역인적법분석비교소서광응전후시망막색소상피층급맥락막상TNF-α단백적표체.결과 광응후1주,면역인적법결과 현시소서맥락막급시망막색소상피층TNF-α표체증가.광응전3 d용약,광응후1주,형광소삼루면적위(0.74±0.33)×104 상소,2주시칙위(0.63±0.20)×104 상소,여대조조[(2.61±0.80)×104 상소]비교균현저감소(t=3.69,3.56;P＜0.05).광응후1주대조조、의나석보조화영리서단항조CNV면적분별위(3.61±0.93)×105、(1.89±0.62)×105화(2.14±0.72)×105상소,여대조조비교,차이균유통계학의의(t=3.10,2.51;P＜0.05.단광응후3 d용약,칙지유의나석보능현저억제CNV병변.조직병이학검사진일보증실,치료조소서적CNV병변직경급후도소우대조조.결론 TNF-α삼여격광유도적CNV적형성.TNF-α억제제능억제격광유도적CNV적형성급삼루.(중화안과잡지,2008,44:200-206)
Objective To investigate the role of TNF-α in the development of laser-induced choroidal neovascularization(CNV)in the mouse.Methods Laser photocoagulation was used to induce CNV in wild-type C57BL/6J mice by making four separate choroidal burns in each eye.Animals were treated 3 days before or after laser injury with recombinant TNF receptor P75(etanercept,5μg/h,groupl,n=12),chimeric monoclonal antibody(infliximab,5μg/h,group2,n=12)for 7 days by intraperitonealy implanted osmotic pumps.PBS was used as control(group3,n=12).The left eyes were removed for histopathologic examination and the right eyes were removed for flatmounts immunohistochemistry immediately after fluorescien angiography.In mice treated with medications 3 days before laser injury,left eyes were collected at 1 or 2 weeks after laser injury.In mice treated with medications 3 days after laser injury,left eyes were collected at 10 days after laser injury.CNV responses were compared by flatmount analysis of CNV-related fluorescence area and by determination of fluorescein angiographic leakage.The level of protein expression of TNF-α was semiquantitatively evaluated by Western blot analysis of the choroidal and RPE layer from mice with or without laser treatment. Results Western blotting demonstrated that TNF-α was highly expressed in choroidal and RPE cells of wild type mice 1 week after laser treatment as compared to the control mice without laser treatment. Etanercept and infliximab administrated 3 days before laser-damage significantly reduced CNV size and pathological fluorescein leakage in comparison to the control group one and two weeks after laser injury. Only etanercept administered 3 days after laser injury still significantly reduced the development of CNV lesions.Histopathological examination confirmed that CNV lesions in treated mice had smaller diameter and thinner center as compared to the control animals.Conclusions Anti-TNF-α treatment reduces the size and leakage of laser-induced CNV.These results suggest the involvement of TNF-α in the development of laser-induced CNV and its potential use as a therapeutic agent in the age related macular degeneration.(Chin J Opbthalmol,2008,44:200-206)
