半乳糖凝集素3抑制剂对大鼠椎间盘软骨终板细胞基质金属蛋白酶-3、趋化因子(C-C基元)配体3及聚蛋白多糖表达的影响

Effect of galectin 3 inhibitor on expression of matrix metalloproteinase-3,chemokine(C-C motif)ligand 3 and aggrecan of rat intervertebral disc cartilage endplate cells

目的探讨半乳糖凝集素3对大鼠椎间盘软骨终板细胞基质金属蛋白酶-3(MMP-3)、趋化因子(C-C基元)配体3(CCL3)、聚蛋白多糖表达的影响。方法将P2代大鼠软骨终板细胞分为2组,一组加入含25μmol/L GB1107半乳糖凝集素3抑制剂(抑制剂组),另一组加入等体积空白溶剂(对照组),采用MTT法检测2组12 h、24 h及48 h细胞的增殖情况;于加入溶剂后24 h收集2组细胞,采用实时荧光定量PCR法检测细胞中MMP-3、CCL3及聚蛋白多糖的mRNA表达水平,采用蛋白质印迹法检测细胞中MMP-3、CCL3及聚蛋白多糖的蛋白表达水平。结果抑制剂组大鼠软骨终板细胞的细胞增殖活性加入抑制剂后12 h开始随时间延长不断降低,加入抑制剂后12 h、24 h、48 h各时间点的细胞增殖活性均低于对照组,差异有统计学意义(P<0.05)。抑制剂组大鼠软骨终板细胞中MMP-3、CCL3及聚蛋白多糖的mRNA和蛋白表达水平均低于对照组,差异有统计学意义(P<0.05)。结论抑制半乳糖凝集素3可降低大鼠终板细胞中MMP-3、CCL3及聚蛋白多糖的表达,提示半乳糖凝集素3可能通过调节细胞外基质的降解、炎性细胞浸润和合成代谢共同影响椎间盘退行性变进程。

Objective To investigate the effect of galectin 3 on expression of matrix metalloproteinase-3(MMP-3),chemokine(C-C motif)ligand 3(CCL3)and aggrecan of rat intervertebral disc cartilage endplate cells.Methods P2 passage rat cartilage endplate cells were divided into 2 groups,one group was added with 25μmol/L GB1107 solvent galectin 3 inhibitor(inhibitor group),and the other group was added with equal volume blank solvent(control group).MTT method was used to detect the proliferation of the cells in 2 groups at 12,24 and 48 h.The mRNA and protein expression levels of MMP-3,CCL3 and aggrecan were detected by real time quantitative PCR and Western blotting at 24 h.Results The proliferation activity of cartilage endplate cells in inhibitor group began to decrease 12 h after adding galectin 3 inhibitor.The proliferation activity of cartilage endplate cells in inhibitor group at 12,24 and 48 h after adding galectin 3 inhibitor was lower than that in the control group,and the difference was statistically significant(P<0.05).The real-time quantitative PCR and Western blotting results showed that the mRNA and protein expression levels of MMP-3,CCL3 and aggrecan in the inhibitor group were also significantly lower than those in the control group,and the differences were statistically significant(P<0.05).Conclusion Inhibition of galectin 3 can decrease the expression of MMP-3,CCL3 and aggrecan in rat cartilage endplate cells,suggesting that galectin 3 may affect the process of intervertebral disc degeneration by regulating the degradation of extracellular matrix,inflammatory cell infiltration and anabolism.