IL-1α对体外培养的牛小梁网细胞MMPs及TIMPs表达的影响

Effect of IL-1α on MMPs and TIMPs expression in cultured bovine trabecular cells

目的观察白细胞介素-1α(IL-1α)对体外培养的牛小梁网细胞分泌基质金属蛋白酶(MMPs)和基质金属蛋白酶组织抑制剂(TIMPs)的影响,探讨IL-1α降低眼压的机制。方法采用酶谱分析法和反向酶谱分析法检测牛小梁网细胞MMP和TIMPs的分泌,并观察IL-1α对MMP和TIMPs分泌的调节作用。结果体外培养的牛小梁细胞分泌MMP-2、MMP-3、MMP-9和TIMP-1;IL-1α对MMP-2、MMP-3、MMP-9的分泌有促进作用,且呈时间和浓度依赖性,其中以IL-1α对MMP-3的分泌促进作用最强;IL-1α对TIMP-1的分泌也有微小的增加作用。结论IL-1α能够显著促进牛小梁网细胞分泌MMPs,打破了MMP/TIMPs的平衡,可能进一步促进小梁网细胞外基质(ECM)的分解,增强房水流动性。

Objective Matrix metalloproteinases （MMPs） are involved in the metabolism of trabecular meshwork extracellular matrix and have been proved to increase aqueous outflow facility. The purpose of this study was to investigate the effects of intedeukin-1α （IL-1α） on the expression of MMPs and tissue inhibitor of metalloproteinases （TIMPs） in cultured bovine trabecular cells. Methods The trabecular meshwork tissue from bovine species was incubated in DMEM medium containing 15% fetal bovine serum and passaged with 0. 125% trypsin and 0. 02% EDTA. The 3rd generation of trabecular meshwork cells of bovine were collected. 500μl of 25 ng/ml IL-1α DMEM medium was added in experimental group,and 500μl of DMEM medium was added in control group. Zymography and reverse-zymography were applied to test secretion of MMPs and TIMPs in cultured bovine trabecular cells. The cultured bovine trabecular cells were treated with IL-1α,and the levels of MMPs and TIMPs in cell conditioned media were assayed by zymography and reverse-zymography. ResuIts The cultured bovine trabecular cells expressed MMP-2, MMP-3, MMP-9 and TIMP-1. Compared with the control group, IL-1α could obviously enhance the expression of MMP-2, MMP-3 and MMP-9 in time-and concentration-dependent manner with the A value 127.31±6.01, 86.49± 6.22 and 73. 00± 5.24 respectively. The production of MMP-3 was maximally activated by 25 ng/ml IL-1α after 72 hours after treatment with the A value 149.38 ± 6.04. IL-1α could increase the expression of TIMP-1 , but the effect was limited. ConcIusion The IL-1α can reduce the intraocular pressure through stimulating the expression of MMPs,breaking the balance of MMPs/TIMPs,eliminating ECM and improving the outflow facility of aqueous.