内源性大麻素AEA对体外培养的牛眼小梁细胞形态和细胞骨架的影响

Effect of endogenous AEA on the morphology and cytoskeleton of bovine trabecular meshwork cells cultured in vitro

目的:研究内源性大麻素(AEA)对体外培养牛眼小梁细胞形态和细胞骨架的影响,探讨大麻素在原发性开角性青光眼(POAG)中的降眼压的机制,为大麻素的眼部用药提供帮助。方法:原代培养的牛眼小梁细胞,经不同质量浓度的AEA作用后,于光学显微镜下观察小梁细胞形态的改变,免疫荧光染色后在荧光显微镜下观察肌动蛋白微丝束的形态改变。免疫组化(SABC法)对a-tubulin(微管蛋白)染色,结果以计算机图像分析系统分析,并统计学检验。结果:经AEA作用后的牛眼小梁细胞,细胞间隙扩大,细胞变圆,细胞立体感增强;肌动蛋白微丝纤维变稀疏,断裂,排列紊乱。各浓度组与对照组比较,微管蛋白的含量减少,其下调呈浓度依赖性。结论:大麻素可改变体外培养的小梁细胞的形态和细胞骨架,引起细胞内的微管蛋白含量的减少,有利于降低小梁网房水引流阻力。

AIM. To study the effect of endogenous AEA on the morphology and cytoskeleton of bovine trabecular meshwork cells cultured in vitro, and to investigate the mechanism of AEA lowering eye pressure in primary open-angle glaucoma （POAG）, in order to provide help in AEA medication in ocular region. METHODS. Primarily cultured bovine trabecular meshwork cells, after treated by different mass concentration AEA, were observed under light microscope to detect cell morphology changes and under fluorescence microscope, after immunofluorescence stain,to detect actin microfllament bundle changes. Immunohistochemical method （SABC） stained a-tubulinthe and the results were analyzed by computer image analysis system and statistically tested. RESULTS. After AEA treatment, bovine trabecular meshwork cell spaces increased, with more rounded and stereoscopic morphology, actin filaments became sparse, disrupted and abnormally aligned. Comparing all concentration groups with control group, microtubulin content was decreased in concentration dependent fashion. CONCLUSION ： Certain mass concentration of AEA may change the morphology and cytoskeleton of trabecular meshwork cells culturedin vitro, induce the decrease of cell microtubulin content, in favor of lowering aqueous fluid drainage resistance in trabecular meshwork.