中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2012年
8期
699-703
,共5页
去瓣型微型角膜刀法准分子激光角膜上皮瓣下磨镶术%准分子激光上皮瓣下角膜磨镶术%紧密连接%Occludin蛋白
去瓣型微型角膜刀法準分子激光角膜上皮瓣下磨鑲術%準分子激光上皮瓣下角膜磨鑲術%緊密連接%Occludin蛋白
거판형미형각막도법준분자격광각막상피판하마양술%준분자격광상피판하각막마양술%긴밀련접%Occludin단백
Flap-free epipolis laser in-situ keratomileusis%Laser-assisted subepithelial keratectomy%Tight junction%Occludin
背景 紧密连接作为角膜上皮屏障的重要构成部分,是角膜屈光手术后角膜上皮损伤修复中的关键因素,目前国内外有关这一方面的研究尚不多见. 目的 探讨和比较去瓣型微型角膜刀法准分子激光角膜上皮瓣下磨镶术(Epi-LASIK)与准分子激光上皮瓣下角膜磨镶术(LASEK)术后早期角膜上皮内紧密连接蛋白occludin的表达.方法 48只健康清洁级成年新西兰白兔按照随机数字表法随机分为2个组,每组24只.2个组均取右眼分别行去瓣型Epi-LASIK和LASEK手术,后者术中也不保留角膜上皮瓣.于术后1、2、3、5d用空气栓塞法处死实验兔并分离角膜组织,每个时间点获取6只兔眼角膜标本.另取2只正常兔为正常对照组.采用免疫荧光检查法观察各组兔眼角膜上皮内occludin蛋白的表达变化,用RT-PCR半定量检测兔眼中央区角膜内occludin mRNA表达的变化.结果 角膜免疫荧光检测结果表明,正常兔眼角膜上皮内可见occludin蛋白表达呈绿色荧光,荧光信号较强且排列有序.术后1～2d,LASEK组兔眼角膜上皮内绿色荧光信号表达强度低,数量少且排列紊乱,而去瓣型Epi-LASIK组角膜上皮内荧光信号强于LASEK组且排列整齐,术后3～5d,2个组荧光信号均增强,排列有序.RT-PCR检测表明,术后1、2、3d,LASEK组兔眼角膜上皮内occludin mRNA表达相对值分别为0.11±0.02、0.25±0.03、0.43 ±0.04,明显低于去瓣型Epi-LASIK组的0.20±0.04、0.44±0.04、0.76±0.04,差异均有统计学意义(t=6.476、12.898、17.315,P＜0.05);术后5d,2个组角膜上皮内occludin mRNA表达相对值差异无统计学意义(t=-0.733,P＞0.05).2个组随着术后时间的延长,occludinmRNA表达相对值均逐渐升高,总体差异有统计学意义(F时间=768.903,P=0.000). 结论 与LASEK比较,去瓣型Epi-LASIK对角膜上皮紧密连接的损害轻,术后角膜上皮紧密连接的修复也更加迅速,提示去瓣型Epi-LASIK有利于减少术后角膜刺激症状和并发症的发生.
揹景 緊密連接作為角膜上皮屏障的重要構成部分,是角膜屈光手術後角膜上皮損傷脩複中的關鍵因素,目前國內外有關這一方麵的研究尚不多見. 目的 探討和比較去瓣型微型角膜刀法準分子激光角膜上皮瓣下磨鑲術(Epi-LASIK)與準分子激光上皮瓣下角膜磨鑲術(LASEK)術後早期角膜上皮內緊密連接蛋白occludin的錶達.方法 48隻健康清潔級成年新西蘭白兔按照隨機數字錶法隨機分為2箇組,每組24隻.2箇組均取右眼分彆行去瓣型Epi-LASIK和LASEK手術,後者術中也不保留角膜上皮瓣.于術後1、2、3、5d用空氣栓塞法處死實驗兔併分離角膜組織,每箇時間點穫取6隻兔眼角膜標本.另取2隻正常兔為正常對照組.採用免疫熒光檢查法觀察各組兔眼角膜上皮內occludin蛋白的錶達變化,用RT-PCR半定量檢測兔眼中央區角膜內occludin mRNA錶達的變化.結果 角膜免疫熒光檢測結果錶明,正常兔眼角膜上皮內可見occludin蛋白錶達呈綠色熒光,熒光信號較彊且排列有序.術後1～2d,LASEK組兔眼角膜上皮內綠色熒光信號錶達彊度低,數量少且排列紊亂,而去瓣型Epi-LASIK組角膜上皮內熒光信號彊于LASEK組且排列整齊,術後3～5d,2箇組熒光信號均增彊,排列有序.RT-PCR檢測錶明,術後1、2、3d,LASEK組兔眼角膜上皮內occludin mRNA錶達相對值分彆為0.11±0.02、0.25±0.03、0.43 ±0.04,明顯低于去瓣型Epi-LASIK組的0.20±0.04、0.44±0.04、0.76±0.04,差異均有統計學意義(t=6.476、12.898、17.315,P＜0.05);術後5d,2箇組角膜上皮內occludin mRNA錶達相對值差異無統計學意義(t=-0.733,P＞0.05).2箇組隨著術後時間的延長,occludinmRNA錶達相對值均逐漸升高,總體差異有統計學意義(F時間=768.903,P=0.000). 結論 與LASEK比較,去瓣型Epi-LASIK對角膜上皮緊密連接的損害輕,術後角膜上皮緊密連接的脩複也更加迅速,提示去瓣型Epi-LASIK有利于減少術後角膜刺激癥狀和併髮癥的髮生.
배경 긴밀련접작위각막상피병장적중요구성부분,시각막굴광수술후각막상피손상수복중적관건인소,목전국내외유관저일방면적연구상불다견. 목적 탐토화비교거판형미형각막도법준분자격광각막상피판하마양술(Epi-LASIK)여준분자격광상피판하각막마양술(LASEK)술후조기각막상피내긴밀련접단백occludin적표체.방법 48지건강청길급성년신서란백토안조수궤수자표법수궤분위2개조,매조24지.2개조균취우안분별행거판형Epi-LASIK화LASEK수술,후자술중야불보류각막상피판.우술후1、2、3、5d용공기전새법처사실험토병분리각막조직,매개시간점획취6지토안각막표본.령취2지정상토위정상대조조.채용면역형광검사법관찰각조토안각막상피내occludin단백적표체변화,용RT-PCR반정량검측토안중앙구각막내occludin mRNA표체적변화.결과 각막면역형광검측결과표명,정상토안각막상피내가견occludin단백표체정록색형광,형광신호교강차배렬유서.술후1～2d,LASEK조토안각막상피내록색형광신호표체강도저,수량소차배렬문란,이거판형Epi-LASIK조각막상피내형광신호강우LASEK조차배렬정제,술후3～5d,2개조형광신호균증강,배렬유서.RT-PCR검측표명,술후1、2、3d,LASEK조토안각막상피내occludin mRNA표체상대치분별위0.11±0.02、0.25±0.03、0.43 ±0.04,명현저우거판형Epi-LASIK조적0.20±0.04、0.44±0.04、0.76±0.04,차이균유통계학의의(t=6.476、12.898、17.315,P＜0.05);술후5d,2개조각막상피내occludin mRNA표체상대치차이무통계학의의(t=-0.733,P＞0.05).2개조수착술후시간적연장,occludinmRNA표체상대치균축점승고,총체차이유통계학의의(F시간=768.903,P=0.000). 결론 여LASEK비교,거판형Epi-LASIK대각막상피긴밀련접적손해경,술후각막상피긴밀련접적수복야경가신속,제시거판형Epi-LASIK유리우감소술후각막자격증상화병발증적발생.
Background Tight junctions are thought to play a significant role in the maintenance of the corneal epithelial defense for the eye,and the restoration of the tight junctions is critical during epithelial wound healing after refractive surgery.However,there are few reports about this study. Objective The aim of this study was to investigate and compare the expression of occludin in corneal epithelium following flap-free epipolis laser insitu keratomileusis (Epi-LASIK) and laser-assisted subepithelial keratectomy ( LASEK ). Methods Forty-eight clean New Zealand white rabbits were randomized into 2 groups,and 24 rabbits for each group.Flap-free Epi-LASIK and LASEK were performed in the right eyes of the rabbits in two groups,and other 2 age matched normal rabbits were as normal controls.The animals were sacrificed and the corneal samples were obtained at 1,2,3,5 days after surgery.Immunofluorescence staining was used to detect the expression of occludin in the corneal epithelium,and RT-PCR was used to identify the level of occludin mRNA in the central cornea. Results Occludin protein was expressed innormal corneal epithelium and showed the green fluorescence with the regular arrangement.The fluorescence intensity was lower in 1 -2 days in LASEK group with the irregular arrangement;while the fluorescence signal in corneal epithelium was stronger in flap-free Epi-LASIK group.3-5 days after surgery,the fluorescence intensities were bothenhanced in two groups.RT-PCR showed that the relative expression of occludin mRNA in corneal epithelium was 0.11 ±0.02,0.25 ± 0.03,0.43 ± 0.04 in LASEK group 1,2,3 days after surgery,respectively,and was lower than those of flap-free Epi-LASIK group ( 0.20± 0.04,0.44 ± 0.04,0.76 ± 0.04 ),showing significant differences between these two groups ( t =6.476,12.898,17.315,P＜ 0.05 ).No significant difference was seen in the expression of occludin mRNA in 5 days after surgery between two groups( t=-0.733,P＞0.05).The relative values of occludin mRNA expression in corneal epithelium were gradually increased with time prolongation,presenting a significant difference among various time points ( Ftime =768.903,P =0.000). Conclusions The reformation of occludin in flap-free Epi-LASIK group is faster than that in LASEK group.Therefore,flap-free Epi-LASIK is prominant in reducing the stimulated symptoms and complication after the surgery.
