体外培养牛视网膜色素上皮细胞激光损伤模型的再探讨

Re-discussion on the laser-injured model of bovine retinal pigment epithelial cells cultured in vitro

目的:探讨体外培养牛视网膜色素上皮细胞激光损伤模型建立的要点,提高模型建立的成功率。方法:实验于2000-05/2001-05在解放军总医院病理科及眼科完成。取三代对数生长期牛视网膜色素上皮细胞随机分为6组加入96孔板中,每组3个样本,用Argon蓝绿激光对视网膜色素上皮细胞进行垂直聚焦照射,激光输出功率各组分别为0,50,100,200,400,800mW,用Argon蓝绿激光对视网膜色素上皮细胞进行垂直聚焦照射;采用甲基噻唑基四唑方法测定各组光吸收值并推算视网膜色素上皮细胞的损伤情况。结果:①激光能量为0时,光吸收值为1.508±0.006。②激光能量为50,100,200,400,800时,光吸收值分别为1.341±0.010,1.303±0.014,1.246±0.024,1.021±0.065,1.027±0.047。③激光能量为50,100,200,400,800时,激光损伤率分别为11.07%,13.59%,17.37%,32.29%,31.90%。④照激光各组均比不照激光组光吸收值下降(P<0.05)。⑤组间比较,随激光能量增大,光吸收值呈逐渐减小趋势(P<0.01)。结论:体外培养牛视网膜色素上皮细胞激光损伤模型的建立是可行的,但是要注意实验参数的选择,才能提高建模成功率。

AIM： To investigate the key point of establishing laser-injured model of bovine retinal pigment epithelial cells cultured in vitro so as to enhance the successful rate of model establishing, METHODS：This experiment was conducted at the Department of Pathology and Department of Ophthalmology, General Hospital of Chinese PLA from May 2000 to May 2001. Three generations of bovine retinal pigment epithelial cells at logarithm growth period were randomly divided into 6 groups and then added into 96-well culture plate, with 3 samples in each group. After Argon laser photocoagulation with different laser powers （0,50,100,200,400,800 roW）on retinal pigment epithelial cells,absorbance was determined with MTT assay and the injury of retinal pigment epithelial cells was calculated. R1ESULTS： ①When the laser power was 0, the absorbance was 1.508±0.006.② When the laser power was 50,100,200,400,800, the absorbance was 1.341±0.010,1.303±0.014,1.246±0.024,1.021±0.065,1.027±0.047, resp ectively. ③ When the laser power was 50,100,200,400,800, the injured rate of laser was 11.07%, 13.59%, 17.37%, 32.29% ,31.90% , respectively . ④The absorbance was decreased in each group with laser photocoagulation as compared with that without laser photocoagulation （P 〈 0.05）. ⑤With the increase of laser power, the absorbance had gradual decreasing trend in the comparison among the groups （P 〈 0.01 ）. CONCLUSION：It is feasible to establish laser-injured model of bovine retinal pigment epithelial cells cultured in vitro, but more attention should be paid on the choice of experimental parameter, so as to enhance the successful rate of model establishing.