摘要
目的:观察肿瘤坏死因子-α对体外培养的牛眼小梁细胞MMP3,MMP9和TIMP-2表达的影响,探讨原发性开角型青光眼的发病机制。方法:对牛眼小梁细胞进行原代及传代培养,应用免疫组化方法(NSE,Ⅷ因子相关抗原染色)鉴定细胞,化学及电子透射电镜对细胞进行形态学及生长特性的观察;对传3代的小梁细胞分别施加含TNF-α终浓度为0,12.5,25,50μg/L的培养液,48h后行MMP3和MMP9免疫组化SP染色,结果进行计算机图像分析并进行统计学检验。分组提取细胞培养液用ELASA检测TIMP-2量的变化。结果:体外培养的牛眼小梁细胞表达MMP3及MMP9,TNF-α可增加MMP3及MMP9的表达,并且抑制TIMP-2的表达。结论:TNF-α在一定范围内可以增加MMP3及MMP9的表达(P<0.05),并抑制TIMP-2的表达(P<0.01),故TNF-α可以减少小梁细胞细胞外基质的堆积,使房水引流通畅,对激光小梁成形术的手术原理有一定的说明意义。
AIM: To study the effect of tumor necrosis factor-α (TNF-α) on the expression of matrix metalloproteinases (MMPs) in the cultured bovine trabecular cells (BTCs)and approach to pathogenesis and mechanism of primary open angle glaucoma (POAG).
METHODS: BTCs were primarily cultured and subcultured. The third passage cells were incubated with different dosages of TNF-α (0, 12.5, 25 and 50μg/L, final concentration, diluted by DMEM) for 48h. MMP3 and MMP9 were determined by immunohistochemistry method. The results were analyzed by the computer image-analysis system and statistical test. Levels of TIMP-2 in cell media were quantified by ELASA.
RESULTS: The cultured BTCs expressed MMP3 and MMP9 . In comparison with the control group, TNF-α increased the expression of MMP3 and MMP9 , but decreased TIMP-2 expression in BTCs.
CONCLUSION: In certain extent, TNF-α can decrease the stacking in ECM and the resistance of aqueous outflow in trabecular meshwork.
出处
《国际眼科杂志》
CAS
2006年第2期328-331,共4页
International Eye Science