CAJ | 학술논문

背景角膜新生血管(CNV)可发生在多种跟表疾病中,常可加重病情,但有效的临床治疗方法仍在探索中. 目的探讨电离辐射对角膜碱烧伤后新生血管的抑制作用.方法 76只清洁级Wistar纯系大鼠中70只用角膜碱烧伤的方法建立大鼠右眼CNV模型,然后按照随机数字表法将造模动物分为β射线10 Gy一次性照射组2只、β射线7 Gy分次照射组17只、β射线10 Gy分次照射组17只、质量分数1％环孢素A(CsA)点眼组17只和角膜碱烧伤模型组17只,6只正常兔6只眼(均取右眼)作为正常对照组.用90 Sr-90Y眼科敷贴器于右眼角膜碱烧伤后第1天开始沿角膜缘进行β射线照射,1％ CsA点眼组用药时间与照射时间相同.实验后每日行裂隙灯检查并计算CNV长度和面积.于角膜碱烧伤后3、5、7d制备角膜组织石蜡切片和匀浆,采用免疫组织化学法检测各组大鼠角膜组织中bcl-2、bax、血管内皮生长因子(VEGF)的表达,分别采用Western blot和逆转录聚合酶链反应(RT-PCR)检测各组大鼠角膜组织中VEGF蛋白及mRNA表达的变化.结果角膜碱烧伤后7d,裂隙灯下可见β射线10 Gy一次性照射组、β射线10 Gy分次照射组均出现角膜溃疡,角膜碱烧伤模型组可见大量CNV生成,而β射线7 Gy分次照射组和1％CsA点眼组CNV明显较少.角膜碱烧伤后7d,β射线7 Gy分次照射组、1％ CsA点眼组CNV长度和面积均明显小于角膜碱烧伤模型组,差异均有统计学意义(长度:q=14.40、24.20,P＜0.01;面积:q=17.80、14.00,P＜0.01).免疫组织化学检测表明,与角膜碱烧伤模型组比较,β射线7Gy分次照射组、1％CsA点眼组大鼠角膜中bcl-2和VEGF蛋白表达均减弱,而bax蛋白表达均增强.RT-PCR检测表明,β射线7 Gy分次照射组、β射线10 Gy分次照射组、1％CsA点眼组VEGF mRNA表达强度明显低于角膜碱烧伤模型组,Western blot检测发现VEGF蛋白的表达与VEGFmRNA的表达遵循同样的规律.结论 90Sr-90Y跟科敷贴器小剂量分次放射治疗可明显抑制角膜碱烧伤后CNV的生长,并且以β射线7 Gy分次照射作用效果最佳. 揹景角膜新生血管(CNV)可髮生在多種跟錶疾病中,常可加重病情,但有效的臨床治療方法仍在探索中. 目的探討電離輻射對角膜堿燒傷後新生血管的抑製作用.方法 76隻清潔級Wistar純繫大鼠中70隻用角膜堿燒傷的方法建立大鼠右眼CNV模型,然後按照隨機數字錶法將造模動物分為β射線10 Gy一次性照射組2隻、β射線7 Gy分次照射組17隻、β射線10 Gy分次照射組17隻、質量分數1％環孢素A(CsA)點眼組17隻和角膜堿燒傷模型組17隻,6隻正常兔6隻眼(均取右眼)作為正常對照組.用90 Sr-90Y眼科敷貼器于右眼角膜堿燒傷後第1天開始沿角膜緣進行β射線照射,1％ CsA點眼組用藥時間與照射時間相同.實驗後每日行裂隙燈檢查併計算CNV長度和麵積.于角膜堿燒傷後3、5、7d製備角膜組織石蠟切片和勻漿,採用免疫組織化學法檢測各組大鼠角膜組織中bcl-2、bax、血管內皮生長因子(VEGF)的錶達,分彆採用Western blot和逆轉錄聚閤酶鏈反應(RT-PCR)檢測各組大鼠角膜組織中VEGF蛋白及mRNA錶達的變化.結果角膜堿燒傷後7d,裂隙燈下可見β射線10 Gy一次性照射組、β射線10 Gy分次照射組均齣現角膜潰瘍,角膜堿燒傷模型組可見大量CNV生成,而β射線7 Gy分次照射組和1％CsA點眼組CNV明顯較少.角膜堿燒傷後7d,β射線7 Gy分次照射組、1％ CsA點眼組CNV長度和麵積均明顯小于角膜堿燒傷模型組,差異均有統計學意義(長度:q=14.40、24.20,P＜0.01;麵積:q=17.80、14.00,P＜0.01).免疫組織化學檢測錶明,與角膜堿燒傷模型組比較,β射線7Gy分次照射組、1％CsA點眼組大鼠角膜中bcl-2和VEGF蛋白錶達均減弱,而bax蛋白錶達均增彊.RT-PCR檢測錶明,β射線7 Gy分次照射組、β射線10 Gy分次照射組、1％CsA點眼組VEGF mRNA錶達彊度明顯低于角膜堿燒傷模型組,Western blot檢測髮現VEGF蛋白的錶達與VEGFmRNA的錶達遵循同樣的規律.結論 90Sr-90Y跟科敷貼器小劑量分次放射治療可明顯抑製角膜堿燒傷後CNV的生長,併且以β射線7 Gy分次照射作用效果最佳.
배경 각막신생혈관(CNV)가발생재다충근표질병중,상가가중병정,단유효적림상치료방법잉재탐색중. 목적 탐토전리복사대각막감소상후신생혈관적억제작용.방법 76지청길급Wistar순계대서중70지용각막감소상적방법건립대서우안CNV모형,연후안조수궤수자표법장조모동물분위β사선10 Gy일차성조사조2지、β사선7 Gy분차조사조17지、β사선10 Gy분차조사조17지、질량분수1％배포소A(CsA)점안조17지화각막감소상모형조17지,6지정상토6지안(균취우안)작위정상대조조.용90 Sr-90Y안과부첩기우우안각막감소상후제1천개시연각막연진행β사선조사,1％ CsA점안조용약시간여조사시간상동.실험후매일행렬극등검사병계산CNV장도화면적.우각막감소상후3、5、7d제비각막조직석사절편화균장,채용면역조직화학법검측각조대서각막조직중bcl-2、bax、혈관내피생장인자(VEGF)적표체,분별채용Western blot화역전록취합매련반응(RT-PCR)검측각조대서각막조직중VEGF단백급mRNA표체적변화.결과 각막감소상후7d,렬극등하가견β사선10 Gy일차성조사조、β사선10 Gy분차조사조균출현각막궤양,각막감소상모형조가견대량CNV생성,이β사선7 Gy분차조사조화1％CsA점안조CNV명현교소.각막감소상후7d,β사선7 Gy분차조사조、1％ CsA점안조CNV장도화면적균명현소우각막감소상모형조,차이균유통계학의의(장도:q=14.40、24.20,P＜0.01;면적:q=17.80、14.00,P＜0.01).면역조직화학검측표명,여각막감소상모형조비교,β사선7Gy분차조사조、1％CsA점안조대서각막중bcl-2화VEGF단백표체균감약,이bax단백표체균증강.RT-PCR검측표명,β사선7 Gy분차조사조、β사선10 Gy분차조사조、1％CsA점안조VEGF mRNA표체강도명현저우각막감소상모형조,Western blot검측발현VEGF단백적표체여VEGFmRNA적표체준순동양적규률.결론 90Sr-90Y근과부첩기소제량분차방사치료가명현억제각막감소상후CNV적생장,병차이β사선7 Gy분차조사작용효과최가.
Background Corneal neovascularization ( CNV ) is a complication of many ocular surface diseases.It often worsen the pathological course.Effective therapy for CNV is still researching. Objective This study was to investigate the inhibitory effect of irradiation on CNV. Methods CNV models were established in 70 right eyes of 70 clean Wistar rats by corneal alkali burning.The models were randomized into β ray 10 Gy once irradiation group( 2 eyes),β ray 7 Gy multiple irradiation group( 17 eyes),β ray 10 Gy multiple irradiation group( 17 eyes),1％ cyclosporin A ( CsA ) eye drops group ( 17 eyes) and model group ( 17 eyes),and 6 matched normal rats were used as normal controls.All treatments started from the first day of the corneal alkali burning.CNV length and area were measured under the slit lamp every day.Corneal samples and homogenate were prepared 3,5,7 days after corneal alkali burning.The expressions of bcl-2,bax,vascular endothelial growth factor(VEGF) in rat corneas were detected by immunochemistry,VEGF proteins and VEGF mRNA were detected by Western blot and reverse transcription polymerase chain reaction (RT-PCR),respectively. Results Corneal ulceration was found in the βray 10 Gy once irradiation group and β ray 10 Gy multiple irradiation group.CNV length and area were much less in the β ray 7 Gy multiple irradiation group and 1％ CsA eye drops group compared with the model group on the seventh day after experiment( length:q=14.40,24.20,P＜0.01 ;area:q=17.80,14.00,P＜0.01 ).Immunochemistry revealed that compared with the model group,expressions of bcl-2 and VEGF proteins were weaker,but the expression of bax protein was stronger in the β ray 7 Gy multiple irradiation group and 1％ CsA eye drops group.RT-PCR showed that the expression of VEGF mRNA in cornea was lower in the β ray 10 Gy multiple irradiation group,β ray 7 Gy multiple irradiation group and 1％ CsA eye drops group in comparison with that in model group,and the results from Western blot showed the same pattern as RT-PCR. Conclusions Low dose irradiation of 90Sr-90Y ophthalmic applicator inhibits CNV formation after alkali burn.The study provide a new understanding of the irradiation for the treatment of CNV.