不同成分灌注液对超声乳化术中兔角膜内皮细胞形态结构的影响

背景:灌注液的成分与其保护角膜内皮功能之间的联系一直是眼科界和制药业研究的热点课题。目的:观察超声乳化术中使用不同灌注液对兔眼角膜内皮细胞结构及功能的影响。设计:随机分组对照动物实验。单位:锦州医学院实验动物中心室,一所市级医院实验动物手术中心实验室。材料:实验于2004-09/2005-03在锦州医学院实验动物中心实验室和锦州市亚东眼科医院实验动物手术中心实验室进行。清洁级3.5月龄纯种日本大耳白兔16只,随机分为4组:正常对照组、生理盐水组、世可组和平衡盐液组,每组4只。灌注液世可(沈阳齐兴制药有限责任公司生产);平衡盐液(美国Alcon公司生产)。方法:用100mL/L水合氯醛(3mL/kg)腹腔麻醉,眼部用4%盐酸奥布卡因滴眼液表面麻醉。行双眼手术。使用美国Alcon超声乳化仪,常规显微手术器械,于上方巩膜缘后2mm作长约3.5mm的巩膜隧道切口,穿刺入前房,注入粘弹剂Viscoat(Alcon公司生产)0.25mL;环行撕囊,直径约5mm;超声乳化头置于中央区乳化吸出晶体核及皮质;术毕关闭切口。于术前和术后6h采用接触性角膜内皮显微镜进行角膜内皮细胞形态学定量测定:角膜内皮细胞密度、细胞面积、六边形细胞百分率和变异系数。术后6h应用锥蓝-茜素红活性染色,光镜下观察各组角膜内皮细胞微细结构的变化。主要观察指标:①角膜内皮细胞形态学定量分析(角膜内皮细胞密度、细胞平均面积、变异系数和六边形细胞百分率)。②角膜内皮细胞形态结构特征。结果:16只大耳白兔均进入结果分析。①术前各组间各项指标比较差异均无显著性。②细胞密度、细胞面积、六边形细胞百分率、变异系数:术后6h生理盐水组、世可组和平衡盐液组均低于正常对照组(P<0.05),世可组和平衡盐液组对比差异无显著性(P>0.05)。③光镜显示生理盐水组角膜内皮细胞形态改变明显,世可组和平衡盐液组角膜内皮细胞改变轻微,无细胞坏死。结论:在超声乳化术中,灌注液对角膜内皮的损伤为化学性损伤,在相同手术条件下,国产灌注液世可对角膜内皮细胞具有良好的保护作用。

Effects of different perfusate on the morphological structure of rabbit corneal endothelium during phacoemulsification

BACKGROUND: The association between the ingredients of perfusate and its protection on corneal endothelium is always the hot issue in ophthalmology and pharmacology.OBJECTIVE: To observe the influence of different perfusates on the structure and function of rabbit corneal endothelium during phacoemulsification.DESIGN: A randomized grouping designed and controlled animal trial.SETTINGS: Laboratory of experimental animal center of Jinzhou Medical College and the laboratory of experimental animal operation of an urban hospital.MATERIALS: The experiments were carried out in the laboratory of experimental animal center of Jinzhou Medical College and the laboratory of experimental animal operation of Jinzhou Yadong Ophthalmology Hospital from September 2004 to March 2005. Sixteen pure Japanese big-ear rabbits of 3.5 months old, clean degree, were randomly divided into four groups with 4 rabbits in each group: normal control group, saline group, shike group and balanced salt solution group. Shike was produced by Shenyang Qixing Pharmaceutical Co.,Ltd.; Balanced salt solution by Alcon Company (USA).METHODS: The rabbits were intraperitoneally anesthetized with 100 ml/L chloral hydrate (3 mL/kg), 4% oxybuprocaine hydrochloride eye drops were used for surface anesthesia of eyes, and both eyes were operated. Alcon phacoemulsification apparatus (USA) and routine microsurgical instruments were used. A 3.5-mm incision was made on sclerotic tunnel at 2 mm posterior to superior limbus of sclera, punctured into the anterior chamber, then 0.25 mL Viscoat (Alcon) was infused. Curvilinear capsulorhexis was performed with the diameter of about 5 mm. The phacoemulsification head was placed in the center to suck out the crystal nucleus and cortex, and the incision was closed after the operation. The morphology of the corneal endothelium was quantitatively determined using contact specular microscope preoperativley and 6 hours postoperatively, including the density and area of corneal endothelium,percent of hexagonal cells and the coefficient of variation. At 6 hours postoperatively, trypan blue-alizarin red active staining was performed, and the changes of slight structures of corneal endothelium were observed under light microscope.MATN OUTCOME MEASURES: ① Quantitative analysis of the forms morphology of corneal endothelium (density and area of corneal endothelium, percent of hexagonal cells and the coefficient of variation); ② Characters of forms and structures of corneal endothelium.RESULTS: All the 16 rabbits were involved in the analysis of results. ① There were no significant differences in the morphological indexes among the four groups preoperatively. ② At 6 hours postoperatively, density and areas of endothelial cells, percent of hexagonal cells and the coefficient of variation were significantly lower in the saline group,shike group and balanced salt solution group than in the normal control group (P ＜ 0.05), and no significant difference was observed between the shike group and balanced salt solution group (P ＞ 0.05). ③ The structural changes of corneal endothelium in the shike group and in balanced salt solution group were alleviated more significantly than those in the saline control group, no necrosis was observed.CONCLUSTON: In phacoemulsification, the damage of perfusate to corneal endothelium is a chemical one. Under the same surgerical conditions, domestic perfusate of shike is as effective as balanced salt solution in protecting endothelial cells.