摘要
目的探讨补骨脂素对破骨细胞分化的影响,以及对破骨细胞ERα、IL-17R基因表达的调节作用,初步阐明补骨脂素抗骨质疏松作用。方法采用核因子κB受体活化因子配体体外诱导小鼠巨噬细胞RAW264.7分化为破骨细胞,以雌二醇作为阳性对照药,补骨脂素分为高剂量(10μmol/L)、中剂量(5μmol/L)、低剂量(2.5μmol/L)。通过TRAP染色细胞数和骨吸收陷窝数评价药物抑制破骨细胞分化作用,同时采用RT-PCR和ELISA检测破骨细胞MMP-9、Cathepsin K、TRAP基因表达和蛋白水平,反映破骨细胞活性,RT-PCR和ELISA检测IL-17R、ER-α的基因表达和蛋白水平,初步探讨补骨脂素作用机制。结果与对照组比较,补骨脂素高、中、低3个浓度均显著抑制破骨细胞形成数和骨陷窝形成数,同时对Cathepsin K、TRAP、IL-17R的基因表达均有显著的抑制作用(P<0.05,P<0.01),并且显著增强ER-α基因表达(P<0.01)。高、中、低剂量的补骨脂素对MMP-9的基因表达有显著的抑制作用(P<0.05)。与对照组比较,高、中剂量的补骨脂素对Cathepsin K的蛋白含量有非常显著的抑制作用(P<0.01);高、中、低剂量的补骨脂素对TRAP、MMP-9、IL-17R的蛋白含量有显著的抑制作用(P<0.05,P<0.01);高、中、低剂量的补骨脂素均显著促进ER-α的蛋白表达(P<0.01)。结论补骨脂素对破骨细胞的形成和溶骨活性均有显著的抑制作用,该作用可能通过提高破骨细胞雌激素受体的表达和抑制IL-17R的表达实现。
Objective To determine the effect of psoralen on osteoclast differentiation and on the regulation of ER-alpha and IL-17 R in the osteoclasts in order to preliminary clarify the anti-osteoporosis effect of psoralen. Methods In order to induce the differentiation of to osteoclasts,mouse macrophage RAW264. 7cells were divided into 5 groups,and treated respectively with receptor activator for nuclear factor-κB ligand（RANKL）,RANKL ＋ estradiol（positive control）,and RANKL ＋ psoralen at 3 different concentrations（10,5 and 2. 5 μmol/L）. In 5 d after the treatment,tartrate-resistant acid phosphatase（TRAP） staining and bone resorption essay were used to identify the inhibitory effects of psoralen on the differentiation of osteoclasts.Transcriptional and protein concentration levels of matrix metallopeptidase 9（MMP-9）,TRAP and Cathepsin K were detected by RT-PCR and ELISA to clarify the activity of osteoclasts,and the expression of IL-17 R and estrogen receptor-α（ER-α） was also detected for the role of psoralen in the process. Results Compared with control cells,high-,middle-and low-dose psoralen significantly inhibited the numbers of osteoclasts and osteoclast-mediated bone resorption,as well as the gene expression of Cathepsin K,TRAP and IL-17R（P〈0. 05,P〈0. 01）,while markedly enhanced the expression of ER-α（P〈0. 01）. The 3 doses of psoralen also reduced the MMP-9 expression significantly（P〈0. 05）. Compared with the control group,the protein level of Cathepsin K was very significantly decreased in the high-and middle-dose psoralen groups（P〈0. 01）.The treatment of psoralen at 3 doses also markedly reduced the protein contents of TRAP,MMP-9 and IL-17R（P〈0. 05,P〈0. 01）,and promoted that of ER-α（P〈0. 01）. Conclusion Psoralen remarkably inhibits the differentiation of osteoclasts and the function of osteolysis at the same time,which might be due to its enhancing the expression of ER and suppressing that of IL-17 R.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2017年第7期641-645,共5页
Journal of Third Military Medical University
基金
国家自然科学基金面上项目(81273816)~~
