JNK抑制剂SP600125对骨质疏松症保护作用的实验研究

Experimental study on the protective effect of JNK inhibitor SP600125 on osteoporosis

目的探讨JNK抑制剂SP600125对骨质疏松症模型成骨细胞和破骨细胞分化及功能的影响。方法使用CCK8试验检测RAW264.7细胞、BMMs细胞和成骨细胞的增殖活性;使用相关染色试验探究JNK抑制剂SP600125的成骨化作用和对破骨细胞分化及功能的影响;使用RT-PCR分别检测BMMs细胞中破骨细胞及成骨细胞特异性基因mRNA表达水平;使用蛋白印迹试验检测RAW264.7细胞中JNK通路和NF-κB通路的活化水平;使用DCFH-DA法检测RAW264.7细胞中ROS水平。结果CCK8试验结果显示,当SP600125浓度≤20μmmol/L,对RAW264.7细胞、BMMs细胞和成骨细胞的增殖活性无显著影响;SP600125不影响成骨细胞钙结节的形成并抑制破骨细胞分化和功能;SP600125抑制的破骨细胞特异性基因的表达但不改变成骨细胞特异性基因的表达;SP600125抑制RANKL诱导的RAW264.7细胞中JNK通路和NF-κB通路的激活及ROS水平的升高。结论JNK抑制剂SP600125能够抑制破骨细胞的分化及骨吸收功能,但对成骨细胞的分化无显著影响。

Objective To investigate the effects of JNK inhibitor SP600125 on the differentiation and function of osteoblasts and osteoclasts in osteoporotic model.Methods CCK8 assay was used to detect the proliferation activity of RAW264.7 cells,BMMs cells and osteoblasts.The ossification of SP600125 and its effect on osteoclast differentiation and function were investigated using staining tests.The mRNA expression levels of osteoclast and osteoblast specific genes in BMMs cells were detected by RT-PCR.Western blot was used to detect the activation levels of JNK and NF-κB pathways in RAW264.7 cells.ROS levels in RAW264.7 cells were detected by DCFH-DA method.Results CCK8 assay result showed that SP600125 had no significant effect on proliferation of RAW264.7 cells,BMMs cells and osteoblasts when SP600125 concentration was less than or equal to 20μM.SP600125 inhibited osteoclast differentiation and function but did not affect the formation of osteoblast calcium nodules.SP600125 inhibited the expression of osteoclast specific genes but did not affect the expression of osteoblast specific genes.SP600125 inhibited the activation of JNK and NF-κB pathways and decreased ROS levels in RANKL-induced RAW264.7 cells.Conclusion JNK inhibitor SP600125 can inhibit osteoclast differentiation and function but has no significant effect on osteoblast differentiation.