摘要
目的观察白细胞介素-1β(IL-1β)对体外培养的人眼小梁细胞基质金属蛋白酶-3(matrix metalloproteinases3,MMP-3)表达的影响。方法采用组织块培养法对人眼小梁细胞(human trabecular cells,HTCs)进行体外原代及传代培养,取传3代小梁细胞分别加入含有IL-1β终浓度为0ng/ml(对照组)、5、10、25、50ng/ml的无血清培养液,24h后采用RT-PCR法及ELISA法检测MMP-3量的变化。结果RT-PCR法检测IL-1β终浓度为5、10、25、50ng/ml实验组MMP-3/GAPDH的相对灰度比值分别为0.3437±0.0764、0.8588±0.0443、1.0079±0.0347、1.3466±0.0309,均高于对照组的相对灰度比值0.1037±0.0494,且各实验组与对照组相比差异有统计学意义(P<0.05);ELISA法检测IL-1β终浓度为5、10、25、50ng/ml实验组的MMP-33浓度值分别为(2.0325±0.2456)ng/ml、(2.4607±0.350)ng/ml、(2.8532±0.1392)ng/ml、(3.5858±0.1141)ng/ml,均高于对照组的浓度值(1.2437±0.3910)ng/ml,且各实验组与对照组相比差异有统计学意义(P<0.05)。结论IL-1β在一定范围内促进MMP-3的表达,并且呈现一定的剂量依赖性,推测IL-1β可以通过促进MMP-3的表达来减少小梁网组织细胞外基质(ECM)的堆积,使房水引流通畅,降低眼内压。
Objective To investigate the effect of Interleukin 1β （ IL-1β ） on the expression of matrix metalloproteinases （MMPs） in cultured human trabecular ceils（HTCs） . Methods Human trabecular cells were primarily cultured and subcultured. The 3rd passage cells were incubated with different dosages of IL-1β（0,5,10 ,25,50 ng/ml, final concentration, diluted by DMEM/F12 without serum ） for 24 hours. MMP-3 were determined by RT-PCR and EL/SA. Results The cultured HTCs expressed MMP-3 . RT-PCR detected that The mean gray scale of MMP-3/GAPDH expressed by the HTCs treated with 0 ng/ml （control） ,5 ng/ml,10 ng/ml,25 ng/ml,50 ng/ml IL-1β for 24h was0. 1037 ±0.0494,0. 3437 ±0.0764,0.8588 ± 0. 0443, 1. 0079 ± 0. 0347,1. 3466 ± 0. 0309, respectively, The difference between these 5 ng/ml, 10ng/ml,25 ng/ml,50ng/ml treated group and that of the control group was statistically sigrfificant. ELISA detected that The constration of MMP-3 expressed by the HTCs treated with 0 ng/ml （control） ,5 ng/ml,10ng/ml,25ng/ml,50ng/ml IL- 1βfor 24h wasl. 2437 ± 0.3910 ng/ml,2.0325 ± 0. 2456 ng/ml,2.4607 ± 0.3207 ng/ml,2. 8532 ± 0. 1392 ng/ml,3. 5858 ± 0.1141 ng/ml, respectively, The difference between these 5 ng/ml, 10ng/ml,25ng/ml,50ng/ml treated group and that of the control group was statistically significant. Conclusion In certain extent, IL-1β can incease expression of MMP-3 in cultured HTCs and increase the degradation of extracellular matrix （ ECM ） localized in trabecular meshwork and decrease the resistance of aqueous outflow, followed by IOP reduction.
出处
《临床眼科杂志》
2008年第4期298-301,共4页
Journal of Clinical Ophthalmology
基金
福建省自然科学基金资助项目(C0510014)
福建医科大学教育学术发展基金资助项目(2006JS0662)
