牛眼外伤性白内障皮质浑浊诱导晶状体上皮细胞凋亡的体外观察

An observation in vitro on the apoptosis of lens epithelial cells induced by opaque lens cortex after traumatic cataract of bovine eyeball

目的观察外伤性白内障皮质浑浊对晶状体上皮细胞（LECs）调亡的影响。方法将牛眼晶状体取出，分为前囊组、后囊组、赤道组和对照组。每组10个晶状体。用1ml注射器针头将晶状体前囊垂直划破3个2mm长切口，并搅动刺破处囊下皮质，制成外伤性白内障模型。然后在杜尔贝科改良伊格尔培养基（Dulbecco’SmodifiedEagle’s medium，DMEM）中分别培养12h、1、3、6、9和12d时观察晶状体变化。取品状体前囊和赤道囊铺片，进行Giemsa染色和原位末端标记（TUNEL）检测，计算LECs凋亡率。结果牛眼晶状体在损伤后12h晶状体皮质出现局限性雾状浑浊；6d时成为完全性浑浊。Giemsa为染色发现随着培养时间的延长，前囊下LECs数量逐渐减少，细胞变小，细胞间隙增宽，边界不清，核固缩逐渐增多。TUNEL染色发现随着皮质浑浊程度的加重和时间的延长，细胞调亡率逐渐增加。对照组无明显变化。结论在体外培养条件下，牛外伤性白内障晶状体皮质浑浊可诱导LECs的凋亡，细胞凋亡率与皮质浑浊的程度和持续时间呈正相关。

Objective To observe the influence of the cortex opacity on the apoptosis of lens epi- thelial cells（ LECs） after the traumatic cataract. Methods The lenses were taken out from the bovine eyes and divided into the anterior capsular group, the posterior capsular group, the equatorial capsular group and the control group. Each group included ten lenses. The capsule of lenses were punctured into three wounds which were about 2mm with a lml disposable needle and the cortex was agitated. Then the lenses were cul- tured in the DMEM culture fluid for 12 h, 1,3,6,9 and 12 d. The lenes were observed and the anterior and equatorial capsule of the lenses were taken out and prepared for Giemsa staining, TUNEL technique and then calculated the rate of apoptosis. Results The bovine lenses appeared local haze and cortex opacity at 12 h after injury and became complete cortex opacity at 6 d. Giemsa staining showed that, with time extension, the number of the epithelial cells on the anterior capsule decreased, the cells became smaller, dilated intercellu- lar space, margin became not distinct, karyopyknosis gradually increased. TUNEL showed that ,with the de- velopment of lens opacity, brown apoptosis cells gradually increased, while the control group showed no sig- nificant change （ P 〈 0.05 ）. Conclusion The lens cortex opacity can induce the apoptosis of LECs in the bovine lens culture traumatic cataract in vitro. The rate of apoptosis is positively correlation to the degree and duration of cortex opacity.