作者：江海鹰， 方林彬， 郑彪， 赖晓宁， 魏建初， 李瑞庄 作者单位：广东医学院附属医院眼科， 广东 湛江 524001
【摘要】 目的：从研究抑制后发性白内障的有效药物入手,探讨术中水分离时应用儿茶素对兔术后晶体上皮细胞增殖的影响及其对兔眼的毒性作用。方法：在兔晶状体超声乳化吸除术中,用1ml不同浓度的儿茶素(50μmol/L、100μmol/L、200μmol/L)在晶状体囊袋内进行水分离,使其直接短暂作用于晶状体上皮细胞。术后随访8周,进行术眼裂隙灯检查、观察眼内压变化及角膜内皮细胞差异,检测术后组织病理学的变化,对儿茶素抑制兔术后晶体上皮细胞增殖效果进行分析。结果：术后2周用药组兔眼未出现后囊膜混浊(posterior capsule opacification,PCO)。术后4周用药组兔眼的PCO明显比对照组眼轻,且随药物浓度增高而减轻。中、低浓度组兔眼的术后炎症反应与对照组比较无明显差异;高浓度组兔眼术后早期出现轻微的毒性反应。组织病理学检查表明,对照组的晶状体上皮细胞增生较用药组明显。结论：①在活体兔中,儿茶素有效地抑制晶状体上皮细胞的增殖,可延缓和防止PCO的形成;②儿茶素对晶体上皮细胞的抑制作用有浓度依赖性;③兔晶状体囊袋内应用50-200μmol/L的儿茶素对角膜内皮细胞没有明显的毒性作用,是防治后发性白内障安全有效的药物。
【关键词】 儿茶素; 后发性白内障; 水分离; 兔
The Inhibitory Effect of EGCG Applied by Intracapsular Method on Rabbit Lens Epithelia Cells: An Experimental Study of After-cataract
JIANG Hai-ying, FANG Lin-bin, ZHENG biao, et al
(The Affiliated Hospital of Guangdong Medical College, Guangdong Zhanjiang 524001, China)
Abstract: Objective: To study the inhibitory effect of green tea extract(-)-Epigallocatechin-3-gallate(EGCG) applied by hydrodissection method on posterior capsule opaci-fication of rabbit lens and evaluate its intraocular toxicity. Method: 1ml of 50μmol/L,100μmol/L,and 200μmol/L EGCG were injected into the capsular bags of the rabbits' lens during the hydrodissection of phacoemulsification, respectively.The drug was exposed to the lens epithelial cells directly and transiently.In the 8-week follow-up after the operation, the posterior capsule opacitication，the change of intraoenlor pressure and the corneal endothelial cell were observed and compared between the control group and the experimental group；The histopathologie changes were also observed．Result: The posterior capsule opacif-ieations only appeared in the control group 4 weeks after the operation．8 weeks after the operation，the posterior capsular opaeification in the experimental group was not mild than that in the control group．The higher the concentration of the drugs，the less the posterior capsular opacification was shown．There was no significant difference between the postoperative inflammation in the control group and the low，middle-concentration experimental group．Slight toxic reaction was observed in the rabbits’eyes of the high-concentration group at early stage after the operation． The histopathologic examination showed that the proliferation of lens epithelial cells in the control group was mole obvious than that in the experimental group．Conclusion: The results showed that EGCG can inhibit the proliferation of lens epithelial cells effectively，significantly prevent the development of PCO in rabbit eyes．Its efficiency is related to the drug's concentration.The toxicity of EGCG to the corneal endothelial cells is not observed, it's a safe and effective durg to prevent the after-cataract when EGCG in concentrations of 50～200umol/L are applied in the capsular bag.
Key words: EGCG; Secondary cataract; Hydrodissection; Rabbits
后发性白内障，即术后晶状体后囊膜混浊(Posterior capsule opacification, PCO)是影响白内障术后视力的并发症，它的发病机理主要为白内障术后残留的晶体上皮细胞过度增殖向后囊移行、化生为成纤维细胞并分泌胶原，使晶体后囊膜混浊［1］。抑制晶体上皮细胞增殖是防治后发性白内障的重要途径。本研究探讨儿茶素作为一种新的抗增殖药短暂应用于兔晶状体超声乳化吸出术中，对兔晶状体后囊膜混浊形成的影响。
1 材料与方法
1.1 实验动物和实验方法:选用健康标准新西兰白兔18只，雌雄兼备，体重1.5～2kg。随机分为三组:每只动物右眼为用药眼，左眼为对照眼。高浓度组(H):兔6只，EGCG浓度为200μmol/L 。中浓度组(M):兔6只，EGCG浓度为100μmol/L。低浓度组(1):兔6只，EGCG浓度为50μmol/L。兔的超声乳化术由同一术者操作，左右眼同一时间依次进行。采用的EGCG的各种浓度按一定的比例用注射用水稀释制成。术前30 min用0.5%复方托品酞胺眼药水3次、1%;阿托品眼药水1次散瞳。盐酸氯胺酮和盐酸氯丙嗪混合液麻醉(4: 1体积混合)，按1.5～2.0 ml/kg体重计算，肌肉注射。手术显微镜下，在上方作宽约3. 2 mm的隧道切口。前房内注入粘弹剂Healon保护角膜内皮和眼前段组织。作直径约3 mm的连续环形撕囊，紧贴前囊下一周作水分离样注入液体1ml，用药眼为不同浓度的EGCG，对照眼为BSS液，上方囊下由辅助切口进针注入液体。使液体存留于晶体囊膜和皮质间5min。完成囊袋内的晶状体超声乳化摘除。用灌注抽吸清除残留的晶状体皮质和药物。结膜下注射庆大霉素2万U、地塞米松1mg。结膜囊涂1%阿托品及妥布霉素地塞米松眼膏。观察无意外予常规喂养。术后观察兔全身情况和眼局部情况，局部滴用新霉素地塞米松眼液，涂妥布霉素地塞米松和阿托品眼膏。
1.2 临床观察方法:术后用裂隙灯观察兔眼前段，直接检眼镜观察眼底。详细记录外眼、结膜、角膜、前房、晶体囊及眼底等情况。术前和术后1～3d用Tonopen-XL型笔式接触眼压计测量眼内压，共4次;于术前和术后2周行角膜内皮检查。
1.3 组织病理学观察:①眼球标本:术后4周及8周用空气注人静脉处死兔后，摘眼球并即放人10%福尔马林固定液中固定24h，常规石蜡包埋切片，行HE染色。光学显微镜下观察，记录并摄片。②晶状体囊标本:为保持晶状体囊及其细胞的完整性，在显微镜下取出完整晶状体囊，相同方法制标本观察。
1.4 统计学方法:采用SPSS8.0统计软件包对手术前后的资料进行方差分析、t检验。
2 结果
2.1 临床观察:术后d1所有手术眼均有不同程度的炎症反应，如角膜水肿、房水混浊。中、低浓度和对照组眼术后3～5d逐渐恢复。高浓度组兔眼术后反应较其他组眼明显，需要10～14d恢复。
2.2 术后晶状体囊混浊情况:术后1周开始，观察兔后囊膜混浊情况。发现术后2周对照组眼晶体后囊出现轻度混浊。低浓度组眼和中浓度组眼分别于术后4周和8周才出现后囊轻度混浊，但均明显轻于对照眼。
表1 手术前后各个浓度组角膜内皮计数（略）
*P<0.05
表2 手术前后用药眼和对照眼角膜内皮损失量的比较（略）
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