目的 探索JNK抑制剂(SP600125)对软骨细胞外基质合成的影响。方法 获取、培养兔耳软骨细胞,通过CCK-8法测定SP600125对细胞增殖的影响。将兔耳软骨细胞接种于PGA/PLA支架,构建细胞-支架复合物,并随机分为实验组与对照组。实验组将细胞-支架复合物置于含有10%FBS和SP600125的DMEM中培养,对照组在含有10%FBS的DMEM中培养。体外培养8周后,RT-PCR检测IGF、COLⅡ、COLⅨ、TGF-β1等软骨特异性基因的表达,并行HE染色与Safranin-O染色。结果 CCK-8法检测显示,相较对照组,实验组软骨细胞的增殖明显增强;体外培养8周后,实验组软骨特异性基因IGF、COLⅡ、COLⅨ、TGFβ1的表达显著下降(P<0.05);组织学观察显示,实验组软骨陷窝的成熟度低,细胞外基质分泌减少。结论 SP600125可降低软骨细胞外基质的合成。

Objective To explore the effect of JNK inhibitor(SP600125)on the synthesis of extracellular matrix of chondrocytes.Methods Rabbit ear chondrocytes were obtained and cultured,and the effect of SP600125 on cell proliferation was determined by CCK-8 method.Rabbit ear chondrocytes were inoculated into PGA/PLA scaffolds,and cell-scaffold complexes were constructed and randomly divided into 2 groups:experimental group and control group.In the experimental group,the cell-scaffold complex was cultured in DMEM containing 10%FBS and SP600125,and the control group was cultured in DMEM containing 10%FBS only.After 8 weeks of in vitro culture,the expression of cartilage-specific genes such as IGF,COLⅡ,COLⅣand TGF-β1 was detected by RT-PCR,and HE staining and Safranin-O staining were performed.Results Compared with control group,the proliferation of chondrocytes in the experimental group was significantly promoted detected by CCK-8.After 8 weeks of in vitro culture,the expression of cartilage-specific genes IGF,COLⅡ,COLⅣand TGF-β1 in the experimental group was significantly lower than that in the control group(P<0.05).Histological observation showed that compared with the control group,the experimental group had a low degree of maturity and a decrease in extracellular matrix secretion.Conclusion SP600125 can reduce the synthesis of extracellular matrix of cartilage.