摘要
目的观察花生四氢酸氨基乙醇(AEA)对体外培养的牛眼小梁网细胞基质金属蛋白酶-3(MMP-3)和基质金属蛋白酶抑制剂-1(TIMP-1)表达的影响。方法对牛眼小梁网细胞进行原代及传代培养,并进行鉴定。对传3代的牛眼小梁网细胞分别施加AEA浓度为0.0、0.1、1.0、10.0μmol·L-1的无血清培养液,作用24 h后提取细胞上清液,并制备10.0μmol·L-1作用6、12、18、24 h的细胞上清液,用酶联免疫吸附测定法检测AEA作用后细胞上清液中MMP-3及TIMP-1水平的变化。结果随着AEA浓度的增加,MMP-3及TIMP-1水平均逐渐增加,并呈剂量依赖性(P<0.05)。10.0μmol·L-1AEA作用6、12、18、24 h后的MMP-3、TIMP-1水平均高于0.0μmol·L-1组(P<0.05),且MMP-3、TIMP-1水平随着作用时间的延长逐渐增加,各时间点之间比较差异有统计学意义(P<0.05)。结论AEA可以促进MMP-3的分泌,早期AEA可以抑制TIMP-1的分泌,进一步打破MMP-3及TIMP-1的平衡,影响细胞外基质的降解,从而起到降低眼压的作用。
Objective To study the effect of anandamide（ AEA） on the expression of matrix metalloproteinase-3（ MMP-3） and tissue inhibitor of metalloproteinase-1（ TIMP-1） in cultured bovine trabecular meshwork（ BTM） cells in vitro.Methods BTM cells were primarily cultured and subcultured. The third passage BTM cells were incubated with different dosages of AEA（ 0. 0,0. 1,1. 0,10. 0 μmol · L- 1,final concentration,diluted by Dulbecco＇s modified Eagle＇s medium） for 24 hours,then the medium were collected. The other third passage BTM cells were incubated with 10. 0 μmol·L- 1AEA,then the medium in 6,12,18,24 h were collected. The concentration of MMP-3 and TIMP-1 in the medium was measured by the enzyme linked immunosorbent assay method. Results With the concentration of AEA increasing,the levels of MMP-3 and TIMP-1 increased,and it showed a dose dependent manner. Compared with 0. 0 μmol·L- 1AEA,the levels of MMP-3 and TIMP-1 in 6,12,18,24 h were higher in 10. 0 μmol·L- 1AEA（ P〈0. 05）; with the prolongation of incubating time,the levels of MMP-3and TIMP-1 increased,and there were significant differences among the different time point（ P〈0. 05）. Conclusion AEA can reduce the intraocular pressure through stimulating the expression of MMP-3,furthermore breaking the balance of MMP-3and TIMP-1,eliminating extracellular matrix to decrease intraocular tension.
出处
《新乡医学院学报》
CAS
2015年第2期123-126,共4页
Journal of Xinxiang Medical University
基金
西安市科技计划基金资助项目(编号:HM1116-3)
