可溶性CD44分子对原发性开角型青光眼患者小梁网细胞凋亡的影响

Effect of soluble CD44 molecules on the apoptosis of cultured trabecular meshwork cells from patients with primary open-angle glaucoma

目的探讨不同浓度的可溶性CD44分子（soluble CD44，sCD44）对原发性开角型青光眼（primary open—angleglaucoma，POAG）患者小梁网细胞凋亡的影响，以及研究sCD44与POAG发病的关系。方法采用组织块培养法原代培养POAG患者小梁网细胞，取传3代的小梁网细胞分别加入含sCD44终浓度为0ng／ml（对照组），1ng／ml，5ng／m]，10ng／ml，25ng／ml，50ng／ml的无血清培养基，分别培养24h后收集细胞，采用CCK-8法、荧光显微镜和流式细胞仪法，研究sCD44对POAG患者小梁网细胞凋亡的影响。结果通过CCK-8法检测发现：随着sCD44终浓度的增加，sCD44对POAG患者小梁网细胞的抑制作用增强，且各实验组与对照组及各实验组之间相比差异有统计学意义（P〈0．05）；通过荧光显微镜观察显示随着sCD44终浓度的增加，早期和晚期凋亡细胞明显增多；通过流式细胞仪法检测sCD44终浓度为1ng／ml，5ng／ml，10ng／ml，25ng，ml，50ng／ml实验组小梁网细胞凋亡率分别为10．73±0．02，17．33±0．03，21．15±0．02，25．13±0．03，30．00±0．03，均高于对照组小梁网细胞凋亡率2．56±0．02，且各实验组与对照组及各实验组之间相比差异有统计学意义（P〈0．05）。结论sCD44在一定浓度范围内可以抑制POAG患者小梁网细胞的增殖，促进细胞的凋亡，并且呈现一定的剂量依赖性，sCD44可能通过作用于小梁网细胞间接参与POAG的发病过程。

Objective To investigate the effect of different concentrations of soluble CD44 mole- cule （sCD44） in trabecular meshwork cells from patients with primary open angle glaucoma （POAG） on the apoptosis of trabecular meshwork cells, to study the relationship between sCD44 and POAG, and further investigate the pathogenesis of POAG. Methods Human trabecular meshwork cells were primarily cultured and subcultured. The 3rd passage cells were incubated with different dosages of sCD44 at （1, 5, 10, 25, 50ng/ml） final concentration, diluted by DMEMJF12 without serum for 24 hours, using CCK-8, fluorescence microscopy and flow cytometry method to study the effect of sCD44 on the apoptosis of cultured trabecular meshwork cells. Results With increasing concentration of sCD44, the inhibition on POAG trabecular meshwork cells were enhanced, by cell counting kit-8 （CCK-8）, and the experimental group compared with the control group and within experimental group was statistically significan （P 〈 0.05 ）. By fluorescence microscopy showed that the early and late apoptotic cells were increased with the increasing final concentration of sCD44. The experi- mental groupOs apoptosis rate of the different dosages of sCD44 at （1, 5, 10, 25, 50ng/ml） final concentration were： 10.72834±0.0223, 17.3267±0.0250, 21.1483±0.0248, 25.1267±0.0281, 29.9900±0.0335, respectively by flow cytometry method, which were higher than the control group of 2.5550±0.0187. The experimental group compared with the control group and within experimental group the difference was statistically significant （P 〈 0.05）. Conclusions In a certain range, sCD44 can inhibit the proliferation of trabecular meshwork cells, cause apoptosis, and show a dose dependent manner. The changing of sCD44 levels may be acting on the trabecular meshwork and indirectly involved in the pathogenesis of POAG.