腺苷对人Müller细胞MMP-2和MMP-9 mRNA表达的影响

EFFECTS OF ADENOSINE ON MMP-2 AND MMP-9 mRNA EXPRESSION IN HUMAN Müller CELL

目的探讨腺苷受体在人Müller细胞中的表达情况以及腺苷(adenosine,ADO)对人Müller细胞中基质金属蛋白酶-2(matrix metalloproteinases-2,MMP-2)和基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)mRNA表达的影响。方法体外培养人Müller细胞系菲尔兹眼科研究所-Müller 1(Moorfields/Institute ofOphthalmology-Müller 1,MIO-M1),采用逆转录-聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)方法检测腺苷受体mRNA在MIO-M1细胞中的表达情况;以0、1、10、100μmol/L浓度的腺苷分别作用MIO-M1细胞24h后,采用逆转录-聚合酶联反应方法检测MMP-2和MMP-9 mRNA的表达情况,对不同浓度组MMP-2和MMP-9 mRNA表达进行单因素方差分析。结果 4种腺苷受体A_1、A_(2A)、A_(2B)和A_3在MIO-M1细胞中均有表达;Real-time PCR结果显示,0、1、10、100μmol/L浓度腺苷作用MIO-M1细胞24 h后,随着腺苷浓度升高,MMP-9 mRNA的表达受到显著抑制(P<0.01,P<0.05),且呈现明显的浓度依赖关系;但各浓度腺苷对MMP-2表达均无明显作用。结论体外培养的人Müller细胞MIO-M1中有腺苷受体A_1、A_(2A)、A_(2B)和A_3表达。腺苷能够抑制MIO-M1细胞中MMP-9 mRNA的表达,并呈现浓度依赖性,但其对MMP-2表达并没有影响。

Objective To identify the presence of adenosine receptors in human MUller cell line MIO -M1 and to investigate the effects of adenosine on matrix metalloproteinases -2 （MMP -2） and matrix metalloproteinases -9 （MMP -9 ） mRNA expression in muller cell. Methods Human Mtiller cell line Moorfields/Institute of Ophthalmology - MUller 1 （ MIO - M1 ） was cultured in Dulbecco modified ealge＇s medium. Adenosine receptors mRNA expression in MIO - M1 ceils was detected by reverse transcription polymerase chain reaction （RT- PCR）. When MIO- M1 cells were treated with different doses of adenosine （0 ~ 100~moL/L） for 24h,the mRNA expression levels of MMP- 2 and MMP-9 in each group was measured by real - time PCR. One - way ANOVA was used to evaluate the difference among different experimental groups. Results There were mRNA expression of four adenosine receptors （A1 ,A2A ,A2B and A3 ） in the cultured human mtiller cell line MIO- M1. The expression of MMP- 9 mRNA in the group treated with different concentrations of adenosine for 24h was significantly decreasedin comparison with the control group （ P 〈 0.01 ）, which showed the concentration dependent effects. However, MMP -2 mRNA expression was not changed by adenosine. Conclusion This study demonstrated the presence of adenosine receptors subtypes （ At , AeA, A2B and A3 ） in human Mailer cell line MIO - M1 at mRNA levels. Adenosine is able to inhibit the expression of MMP - 9 mRNA in MIO - M1 cells in a concentration - dependent manner, but could not affect the expression of MMP - 2 mRNA.