中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2013年
6期
529-534
,共6页
李婧%沈政伟%李德忠%叶娅%姜黎%尹禾%薛林平
李婧%瀋政偉%李德忠%葉婭%薑黎%尹禾%薛林平
리청%침정위%리덕충%협아%강려%윤화%설림평
去上皮瓣微型角膜刀法角膜上皮瓣下原位磨镶术%Bevacizumab%雾状混浊%基底膜%转化生长因子-β1%α-平滑肌肌动蛋白
去上皮瓣微型角膜刀法角膜上皮瓣下原位磨鑲術%Bevacizumab%霧狀混濁%基底膜%轉化生長因子-β1%α-平滑肌肌動蛋白
거상피판미형각막도법각막상피판하원위마양술%Bevacizumab%무상혼탁%기저막%전화생장인자-β1%α-평활기기동단백
Off-flap epipolis laser in situkeratomileusis%Bevacizumab%Haze%Basement membrane%Transforming grown facter-β1%α-Smooth muscle actin
背景 角膜屈光手术后角膜上皮下雾状混浊(haze)是影响术后视力改善的主要原因,转化生长因子-β1(TGF-β1)及α-平滑肌肌动蛋白(α-SMA)均参与haze的形成过程.研究发现bevacizumab除可抑制角膜新生血管外,还可加快角膜上皮基底膜形成,但目前bevacizumab对准分子激光角膜屈光手术后haze的产生是否有影响及其机制尚不明确. 目的 探讨bevacizumab对兔去上皮瓣微型角膜刀法角膜上皮瓣下原位磨镶术(Off-flap Epi-LASIK)术后术区TGF-β1和α-SMA表达的影响.方法 24只普通级健康青紫蓝兔均行右眼Off-flap Epi-LASIK手术,术后按随机数字表法随机分为3个组,每组8只.术后10 min,2个组兔眼结膜下注射bevacizumab 2.5 mg(0.1 ml),其中bevacizumab重复注射组兔眼于术后4d再次结膜下注射bevacizumab 2.5 mg(0.1 ml).生理盐水组兔眼术后以同样的方法给予0.1ml生理盐水；另选2只正常兔作为空白对照组.术后每日裂隙灯显微镜下观察haze程度并参照SundarRayde的标准进行评分,分别于术后1周、4周取4只兔眼角膜组织行苏木精-伊红染色和过碘酸希夫染色,观察角膜组织的病理学变化；采用免疫组织化学法检测角膜组织中TGF-β1、α-SMA的表达. 结果 术后3个组兔眼角膜上皮均于术后4～5d完全愈合.术后1周、4周bevacizumab单次注射组和bevacizumab重复注射组兔眼haze积分值均明显低于生理盐水组,差异均有统计学意义(P＜0.05),但bevacizumab单次注射组和bevacizumab重复注射组间差异均无统计学意义(P＞0.05).术后4周,角膜组织病理学检查显示bevacizumab单次注射组和bevacizumab重复注射组角膜纤维化反应少,角膜创伤愈合比生理盐水组快.术后1周bevacizumab单次注射组和bevacizumab重复注射组角膜基质层TGF-β1的表达量分别为(49.8±2.1)PU和(38.6±4.4) PU,4周时分别为(37.7±4.8) PU和(28.3±3.5)PU,各时间点间TGF-β1的表达量均明显低于生理盐水组的(65.1±5.3) PU和(51.6±2.2) PU,差异均有统计学意义(P＜0.05).术后1周bevacizumab单次注射组和bevacizumab重复注射组角膜基质层α-SMA的表达量分别为(67.2±10.0) PU和(32.7±3.1)PU,4周时分别为(34.2±5.7) PU和(22.8±3.0) PU,各时间点间α-SMA的表达量均明显低于生理盐水组的(87.8±7.7) PU和(59.4±5.6) PU,差异均有统计学意义(P＜0.05).各时间点间bevacizumab重复注射组角膜基质中TGF-β1和α-SMA的表达量均明显低于bevacizumab单次注射组,差异有统计学意义(P＜0.01).过碘酸希夫染色表明,术后早期bevacizumab单次注射组和bevacizumab重复注射组角膜基底膜比生理盐水组连续和完整. 结论 Bevacizumab结膜下注射能下调Off-flap Epi-LASIK术后兔眼早期角膜中TGF-β1和α-SMA的表达水平,减少haze的形成.
揹景 角膜屈光手術後角膜上皮下霧狀混濁(haze)是影響術後視力改善的主要原因,轉化生長因子-β1(TGF-β1)及α-平滑肌肌動蛋白(α-SMA)均參與haze的形成過程.研究髮現bevacizumab除可抑製角膜新生血管外,還可加快角膜上皮基底膜形成,但目前bevacizumab對準分子激光角膜屈光手術後haze的產生是否有影響及其機製尚不明確. 目的 探討bevacizumab對兔去上皮瓣微型角膜刀法角膜上皮瓣下原位磨鑲術(Off-flap Epi-LASIK)術後術區TGF-β1和α-SMA錶達的影響.方法 24隻普通級健康青紫藍兔均行右眼Off-flap Epi-LASIK手術,術後按隨機數字錶法隨機分為3箇組,每組8隻.術後10 min,2箇組兔眼結膜下註射bevacizumab 2.5 mg(0.1 ml),其中bevacizumab重複註射組兔眼于術後4d再次結膜下註射bevacizumab 2.5 mg(0.1 ml).生理鹽水組兔眼術後以同樣的方法給予0.1ml生理鹽水；另選2隻正常兔作為空白對照組.術後每日裂隙燈顯微鏡下觀察haze程度併參照SundarRayde的標準進行評分,分彆于術後1週、4週取4隻兔眼角膜組織行囌木精-伊紅染色和過碘痠希伕染色,觀察角膜組織的病理學變化；採用免疫組織化學法檢測角膜組織中TGF-β1、α-SMA的錶達. 結果 術後3箇組兔眼角膜上皮均于術後4～5d完全愈閤.術後1週、4週bevacizumab單次註射組和bevacizumab重複註射組兔眼haze積分值均明顯低于生理鹽水組,差異均有統計學意義(P＜0.05),但bevacizumab單次註射組和bevacizumab重複註射組間差異均無統計學意義(P＞0.05).術後4週,角膜組織病理學檢查顯示bevacizumab單次註射組和bevacizumab重複註射組角膜纖維化反應少,角膜創傷愈閤比生理鹽水組快.術後1週bevacizumab單次註射組和bevacizumab重複註射組角膜基質層TGF-β1的錶達量分彆為(49.8±2.1)PU和(38.6±4.4) PU,4週時分彆為(37.7±4.8) PU和(28.3±3.5)PU,各時間點間TGF-β1的錶達量均明顯低于生理鹽水組的(65.1±5.3) PU和(51.6±2.2) PU,差異均有統計學意義(P＜0.05).術後1週bevacizumab單次註射組和bevacizumab重複註射組角膜基質層α-SMA的錶達量分彆為(67.2±10.0) PU和(32.7±3.1)PU,4週時分彆為(34.2±5.7) PU和(22.8±3.0) PU,各時間點間α-SMA的錶達量均明顯低于生理鹽水組的(87.8±7.7) PU和(59.4±5.6) PU,差異均有統計學意義(P＜0.05).各時間點間bevacizumab重複註射組角膜基質中TGF-β1和α-SMA的錶達量均明顯低于bevacizumab單次註射組,差異有統計學意義(P＜0.01).過碘痠希伕染色錶明,術後早期bevacizumab單次註射組和bevacizumab重複註射組角膜基底膜比生理鹽水組連續和完整. 結論 Bevacizumab結膜下註射能下調Off-flap Epi-LASIK術後兔眼早期角膜中TGF-β1和α-SMA的錶達水平,減少haze的形成.
배경 각막굴광수술후각막상피하무상혼탁(haze)시영향술후시력개선적주요원인,전화생장인자-β1(TGF-β1)급α-평활기기동단백(α-SMA)균삼여haze적형성과정.연구발현bevacizumab제가억제각막신생혈관외,환가가쾌각막상피기저막형성,단목전bevacizumab대준분자격광각막굴광수술후haze적산생시부유영향급기궤제상불명학. 목적 탐토bevacizumab대토거상피판미형각막도법각막상피판하원위마양술(Off-flap Epi-LASIK)술후술구TGF-β1화α-SMA표체적영향.방법 24지보통급건강청자람토균행우안Off-flap Epi-LASIK수술,술후안수궤수자표법수궤분위3개조,매조8지.술후10 min,2개조토안결막하주사bevacizumab 2.5 mg(0.1 ml),기중bevacizumab중복주사조토안우술후4d재차결막하주사bevacizumab 2.5 mg(0.1 ml).생리염수조토안술후이동양적방법급여0.1ml생리염수；령선2지정상토작위공백대조조.술후매일렬극등현미경하관찰haze정도병삼조SundarRayde적표준진행평분,분별우술후1주、4주취4지토안각막조직행소목정-이홍염색화과전산희부염색,관찰각막조직적병이학변화；채용면역조직화학법검측각막조직중TGF-β1、α-SMA적표체. 결과 술후3개조토안각막상피균우술후4～5d완전유합.술후1주、4주bevacizumab단차주사조화bevacizumab중복주사조토안haze적분치균명현저우생리염수조,차이균유통계학의의(P＜0.05),단bevacizumab단차주사조화bevacizumab중복주사조간차이균무통계학의의(P＞0.05).술후4주,각막조직병이학검사현시bevacizumab단차주사조화bevacizumab중복주사조각막섬유화반응소,각막창상유합비생리염수조쾌.술후1주bevacizumab단차주사조화bevacizumab중복주사조각막기질층TGF-β1적표체량분별위(49.8±2.1)PU화(38.6±4.4) PU,4주시분별위(37.7±4.8) PU화(28.3±3.5)PU,각시간점간TGF-β1적표체량균명현저우생리염수조적(65.1±5.3) PU화(51.6±2.2) PU,차이균유통계학의의(P＜0.05).술후1주bevacizumab단차주사조화bevacizumab중복주사조각막기질층α-SMA적표체량분별위(67.2±10.0) PU화(32.7±3.1)PU,4주시분별위(34.2±5.7) PU화(22.8±3.0) PU,각시간점간α-SMA적표체량균명현저우생리염수조적(87.8±7.7) PU화(59.4±5.6) PU,차이균유통계학의의(P＜0.05).각시간점간bevacizumab중복주사조각막기질중TGF-β1화α-SMA적표체량균명현저우bevacizumab단차주사조,차이유통계학의의(P＜0.01).과전산희부염색표명,술후조기bevacizumab단차주사조화bevacizumab중복주사조각막기저막비생리염수조련속화완정. 결론 Bevacizumab결막하주사능하조Off-flap Epi-LASIK술후토안조기각막중TGF-β1화α-SMA적표체수평,감소haze적형성.
Background Haze formation is a key factor of vision reduce following corneal refractive surgery.Transforming growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) are documented to participate in haze formation.Laboratory study showed that bevacizumab can not only inhibit corneal neovascularization,but also promote the healing of corneal epithelial basement membrane.However,the impact of bevacizumab on corneal healing after Off-flap epipolis laser in situkeratomileusis (Off-flap Epi-LASIK) is unclear.Objective The present study was to investigate the inhibition effect of bevacizumab on corneal haze after off-flap Epi-LASIK and its active mechanism.Methods Off-flap Epi-LASIK was performed in 24 adult pigmented rabbits and these rabbits were randomized into three groups.Bevacizumab of 0.1 ml (2.5 mg) was subconjunctivally injected 10 minutes after surgery in 16 rabbits and the same amount of bevacizumab was repeatedly injected 4 days after the initial injection in 8 eyes of 16 eyes.In addition,equivalent amount of normal saline solution was used in the same way in the other 8 rabbits.Another 2 health rabbits were used as the blank controls.Operative eyes were examined by slit lamp biomicroscope daily after surgery and haze was scored based on SundarRayde criteria.Corneas were obtained 4 weeks after operation for hematoxylin & eosin and periodic acid Schiff staining.Expressions of TGF-β1 and α-SMA in corneal tissue were detected by immunochemistry.Results Corneal epithelium healed completely in all eyes 4-5 days after operation.The haze scores were lower in the bevacizumab single injection group and repeat injection group than those in the normal saline solution group (P＜0.05) in 1 week and 4 weeks after operation.However,no significant difference was seen in the haze scores between the bevacizumab single injection group and repeat injection group (P ＞ 0.05).The hostopathological examination showed that the fibrosis response of cornea tissue was slight in the bevacizumab single injection group and repeat injection group comparison with the normal saline solution group.At 1 week after operation,the expression levels of TGF-β1 were (49.8 ± 2.1) PU and (38.6 ±4.4) PU in the bevacizumab single injection group and repeat injection group,and those of 4 weeks were (37.7 ±4.8) PU and (28.3 ± 3.5) PU,indicating significant decrease in the TGF-β1 expression compared with (65.1 ±5.3) PU and (51.6±2.2) PU of the normal saline solution group in both 1 week and 4 weeks (P＜0.01).The expression levels of α-SMA in corneas were (67.2±10.0) PU and (32.7±3.1) PU at 1 week,and (34.2±5.7) PU and (22.8±3.0) PU at4 weeks after operation in the bevacizumab single injection group and repeat injection group,which were significantly lower than (87.8±7.7) PU and (59.4±5.6) PU in the normal saline solution group in both 1 week and 4 weeks (P＜0.05).Meanwhile,the expression levels of TGF-β1 and α-SMA were declined in the bevacizumab repeat injection group compared with single injection group (P＜0.01).Periodic acid Schiff staining exhibited that the basement membrane of cornea was intact and continued in bevacizumab injection group.But corneal basement membrane was discontinuous in the normal saline solution group.Conclusions Subconjunctival injection of bevacizumab downregulates the expressions of TGF-β1and α-SMA in cornea after Off-flap Epi-LASIK and thus prevents haze formation.
