小白菊内酯对核因子κB受体活化因子配体诱导的RAW264．7细胞向破骨细胞分化的影响  

Effects of parthenolide on osteoclast differentiation of RAW264. 7 cell induced by receptor activator of nuclear factor κB ligand

作　　者：吴侗[1] 孙晓麟[2] 杜燕[2] 龙丽[1] 张晶[1] 苏江[1] 周乔[1] 周彬[1] 

机构地区：[1]四川省医学科学院 四川省人民医院风湿免疫科,成都610072 [2]北京大学人民医院风湿病研究所

出　　处：《中华风湿病学杂志》2015年第7期468-472,共5页Chinese Journal of Rheumatology

摘　　要：目的研究小白菊内酯对NF-κB受体活化因子配体（RANKL）诱导的RAW264．7细胞向破骨细胞分化的影响。方法以RANKL单独诱导小鼠单核巨噬细胞RAW264．7细胞为对照组，分别采用不同浓度（0．5、1、2um01／L）Zb白菊内酯联合RANKL诱导RAW264．7细胞分化培养。第3、5、7天采用抗酒石酸酸性磷酸酶（TRAP）染色法检测破骨细胞样细胞并计数；ELISA法检测第7天各组培养液上清中抗酒石酸酸性磷酸酶5b（TRAP5b）含量；实时（RT）．PCR检测第7天各组破骨细胞标志基因降钙素受体（CTR）和MMP-9的表达。使用SPSS17．0统计学软件进行方差分析和t检验以比较组间差异。结果不同的培养条件下RANKL均能诱导RAW264．7细胞分化为成熟的破骨细胞。与同期对照组比较，诱导第3、5、7天，小白菊内酯各组破骨细胞数均明显减少；随小白菊内酯浓度的增大而破骨细胞数减少幅度更显著，0．5、1、2μmol／L各组第7天破骨细胞数较对照组分别下降为36．3％、40．8％、49．3％，细胞数差异有统计学意义（f=7．758，8．742，10．56；P〈0．05）。各组第7天培养液中TRAP5b含量变化与细胞计数结果相符（P〈0．05）。TRAP阳性破骨细胞的CTR、MMP-9表达量随着小白菊内酯浓度增加而减少，2μmol／L小白菊内酯组最低，与对照组比较小白菊内酯0．5、1、2μmol／L各组差异均有统计学意义（P〈0．05）。结论小白菊内酯对RANKL诱导下RAW264．7细胞向破骨细胞的分化具有抑制作用，且抑制作用具有剂量依赖性．Objective To study the effects of parthenolide on osteoclast differentiation of RAW264. 7 cell induced by receptor activator of nuclear factor κB ligand （RANKL）. Methods The mouse macrophage RAW264.7 cells induced by RANKL was used alone as the control group, different concentrations of par- thenolide （0.5, 1, 2μmol/L） were added to culture the RAW264.7 cells. On the third, fifth and seventh day, the tartrate resistant acid phosphatase （TRAP） staining method was used to detect osteodast-like cells and the cell number was count; the contents of tartrate resistant acid phosphatase （TRAP5b） in the Culture supernatant of each groups were detected by enzyme linked immunosorbent assay （ELISA） and the expression of osteodast marker gene alcitonin receptor （CTR） and matrix metalloproteinase （MMP）-9 in each groups were detected by reahime-polymerase chain reaction （PCR） on the seventh day. We use Chi-square test and t test to test the differences between groups by SPSS 17.0. Results In different culture conditions, RANKL could always induce the RAW264.7 cell differentiate into mature osteoclasts. Compared with the control group at the same time control group, on the third, fifth and seventh day, he number of mature osteoclasts induced were obviously decreased in groups adding different concentration of PAR; the number of induced osteoclasts decreased along with the increase of parthenolide concentration, on the seventh day in 0.5, 1, 2 μmol/L concentration PAR groups, the number of mature osteoclasts compared with the control group were descended 36.3%, 40.8%, 49.3% （t=7.758, 8.742, 10.56;P〈0.05）; the contents of TRAP5b in the culture supernatant were consistent with the cell counting results on the seventh day （P〈0.05）. The expression of CTR and MMP-9 by TRAP positive osteoelasts decreased along with the increase of parthenolide concentration, and the 2 Ixmol/L group was the lowest. Compared with the control group, there were statistically significant differences with the di

关 键 词：破骨细胞 小白菊内酯 核因子ΚB受体活化因子配体 抗酒石酸酸性磷酸酶染色 

分 类 号：R285[医药卫生—中药学]