NS398对白介素1α诱导兔角膜基质细胞环氧化酶2表达的抑制  

NS398 inhibits expression of cyclooxygenase-2 induced by IL-1α in rabbit corneal stromal cells

作　　者：赵桂秋[1] 胡丽婷[1] 姜楠[1] 车成业[1] 

机构地区：[1]青岛大学医学院附属医院眼科,266003

出　　处：《眼科研究》2009年第9期760-764,共5页Chinese Ophthalmic Research

摘　　要：目的探讨氮-2,环己氧-4,硝基苯-甲基磺胺(NS398)对白介素1α(IL-1α)诱导的兔角膜基质细胞环氧化酶2(COX-2)表达的影响。方法体外培养兔角膜基质细胞,实验组分别以含0、25、50、100、200μmol/L NS398的培养液孵育2 h后加入IL-1α诱导COX-2表达,24 h后实时荧光定量聚合酶链反应(Real-Time PCR)检测兔角膜基质细胞中COX-2基因表达的差异,四甲基偶氮唑盐(MTT)比色法检测不同浓度NS398对细胞生长的影响,并与对照组进行比较。结果Real-Time PCR结果显示IL-1α诱导后24 h各组COX-2 mRNA表达量差异有统计学意义(F=988.45,P<0.01);对照组与各实验组以及各实验组之间COX-2 mRNA表达量进行多重比较可见,0μmol/LNS398浓度组与对照组之间差异无统计学意义(q=1.332 2,P>0.05),其他NS398浓度组与对照组相比差异均有统计学意义(q=34.789 6,48.329 8,65.801 0,70.813 1,P<0.01),随培养液中NS398浓度的增加,IL-1α刺激后兔角膜基质细胞中COX-2 mRNA表达量不断下降(q=36.121 8,49.662 0,67.133 2,72.145 3,13.540 2,31.011 4,36.023 5,17.471 2,22.483 3,5.012 1,P<0.01);MTT法检测结果显示,100μmol/L、200μmol/L的NS398对兔角膜基质细胞生长均有明显的抑制作用(q=12.769 3,20.908 7,P<0.01)。结论NS398能够有效抑制IL-1α诱导的兔角膜基质细胞COX-2的表达,但超过一定浓度会对细胞产生明显毒性作用。Objective Research demonstrated that cyelooxygenase-2 （COX-2）promote the pathogenesis and development of keratitis. NS398, a non-steroidal anti-inflammatory drug, is a selectivity inhibitor of COX-2. It plays an important role in decreasing the expression of COX-2 in inflammatory cornea. This study was designed to research the effect of NS398 on the expression of COX-2 induced by IL-1α in rabbit corneal stromal ceils in vitro. Methods Rabbit corneal stromal cells were cultured in vitro. In experimental groups,the cells grew in DMEM containing 10% fetal bovine serum using standard techniques and treated with NS398 at the dose of 0,25,50,100 and 200 μmol/L respectively for 2 hours and then were stimulated by IL-1α The rabbit corneal stromal ceils cultured in DMEM containing 0. 1% DMSO were used as control. After 24 hours, Real-Time polymerase chain reaction（ Real-Time PCR） was used to examine the expression of COX-2 in cultured cells at mRNA level,and methyl thiazol tetrazolium（MTT） rapid photocolorimetric assay was used to determine the proliferative activity of cells treated with different doses of NS398. Results The optimal culture concentration of NS398 for expression of COX2 and growth of corneal stromal cells was 25 μmol/L and 50μ mol/L. The cultured cells presented the alteration of cellular shape in 100 and 200 μmol/L NS398 treated groups under the inverted microscope and showed the inhibition of cells growth and proliferation. Real-Time PCR showed that the related expression of COX-2 mRNA among all the groups was statistically significantly different after stimulated for 24 hours by IL-1α （F = 988.45, P 〈 0.01 ）, and the expressing level was gradually declinded with the increase of NS398 concentration,showing a considerabally intergroup difference（ P 〈 0. 01 ） . The result of MTT assay showed that the OD value of cultured cells was significantly different among all groups （F = 72.66,P 〈 0. 01 ） and showed significant diffefence between 100 and 200 μ mol/L NS398 treated

关 键 词：NS398 环氧化酶2 白介素1Α 角膜基质细胞 

分 类 号：R772.2[医药卫生—眼科] R988.1[医药卫生—临床医学]