IL-1β对兔关节软骨细胞MMP-1/-13mRNA表达和NO的影响

Effects of interleulin-1β on gene expression of metalloproteinase-1/-13 mRNA and nitric oxide in rabbit articular chondrocytes

目的观察白介素-1β(IL-1β)对体外培养的兔关节软骨细胞基质金属蛋白酶-1/-13(MMP-1/-13)基因表达的调节作用和对一氧化氮(NO)的影响,以了解骨关节炎(osteoarthritis,OA)软骨损伤的机制。方法将体外培养的兔关节软骨细胞随机分为5组,每组有3个样本,第1组为空白对照组,第2,3,4,5组分别加入10ng/mL的IL-1β,作用时间分别为8,16,24,36h。采用实时荧光定量PCR(real-timePCR)的方法,观察兔关节软骨细胞MMP-1/-13基因的表达情况,以空白组为对照,比较不同时间段软骨细胞MMP-1/-13基因表达,并收取培养细胞上清液测定一氧化氮(NO)含量,比较各组变化。结果正常兔关节软骨细胞具有MMP-1/-13基因表达,但是表达量较低,而各实验组MMP-1/-13mRNA都明显增高。尤其是MMP-1mRNA增高明显,在36h内随着时间的延长逐渐增高,且各组间差异具有统计学意义(P<0.05);MMP-13mRNA增高后维持在高水平,但各组间无明显差异。NO的含量也随着时间的延长而逐渐增高。结论IL-β1随着时间延长,正向调节兔关节软骨细胞MMP-1/-13mRNA表达,并能促进NO的释放增加,从而使它们在OA软骨破坏的机制中发挥重要作用,但对MMP-1/-13具体调节机制不尽相同。

Objective To observe the effect of IL-1β in regulating the expression of matrix metalloproteinase-1/-13mRNA and nitric oxide（NO）in rabbit articular chondrocytes.Methods Rabbit articular chondrocytes were cultivated in vitro and randomly divided into 5 groups,each group was stimulated with 10 ng/mL IL-1β at different time（0,8,16,24,36 h separately）.Real-time PCR was used to observe the MMP-1/-13 mRNA expression in each group.Medium were collected to assess the levels of NO.Results There was expression of MMP-1/-13mRNA in normal rabbit chondrocytes although the expression level was low.In comparison with that of normal controls,IL-1β could increase the MMP-1/-13 mRNA expression and NO levels in medium,especially MMP-1mRNA.MMP-1/13 and NO increased as the time extended,and the differences among groups were significant（P〈0.05）.While MMP-13mRNA levels increased and subsequently remained stable.Conclusions IL-1β can increase MMP-1/-13mRNA expression and NO levels in rabbit articular chondrocytes.IL-1β or derived MMPs and NO appears to serve an important role in the pathogenesis of osteoarthritis（OA）,and the regulating mechanism between MMP-1/-13 may be different.