柚皮素对破骨细胞特异性基因及OPG/RANKL/RANK表达的影响

目的 观察柚皮素对破骨细胞特异性基因组织蛋白酶-K(CATK)、基质金属蛋白酶-9(MMP-9)、抗酒石酸酸性磷酸酶(TRAP)及骨保护素/核因子-κB受体活化因子配体/核因子-κB受体活化因子(OPG/RANKL/RANK)表达的影响.方法 采用全骨髓细胞诱导法培养破骨细胞,细胞分5组:空白对照组、HG-DMEM诱导组、HG-DMEM+0构.1μmol/L柚皮素组、HG-DMEM+1μmol/L柚皮素组、HG-DMEM+10μmol/L柚皮素组.HE染色、TRAP染色观察破骨细胞形态;分光光度计检测TRAP阳性细胞数;Real time-PCR和Western-blot检测CATK、MMP-9、TRAP及OPG/RANKL/RANK mRNA和蛋白表达.结果 与空白对照组比较,HG-DMEM诱导组TRAP阳性细胞数、TRAP、MMP-9、CATK、RANKL、RANK mRNA和蛋白表达明显增加,OPG mRNA和蛋白表达明显降低(P<0.05).与HG-DMEM诱导组比较,柚皮素预处理24 h能够明显降低TRAP阳性细胞数、CATK、MMP-9、TRAP、RANKL、RANK mRNA和蛋白表达,增加OPG mRNA和蛋白表达(P<0.05),呈剂量依赖性.结论 柚皮素能够抑制破骨细胞的生成和分化,该作用可能是通过OPG/RANKL/RANK信号通路抑制TRAP、MMP-9、CATK表达实现的.

Effects of naringenin on the expressions of osteoclast specific genes and OPG/RANKL/RANK in osteoclasts

Objective To observe the effects of naringenin on the expressions osteoclast specific genes -CATK,MMP-9,TRAP and OPG/RANKL/RANK in osteoclasts.Methods The osteoclasts were cultured by whole bone marrow cells induction method.The experiment was divided into 5 groups:blank control group,HG-DMEM induction group,HG-DMEM+naringenin (0.1μmol/L) group,HG-DMEM +naringenin (1μmol/L) group and HG-DMEM +naringenin (10μmol/L) group.After HE and TRAP stainings , the cell morphous was observed under inverted microscope .The TRAP positive cell number was detected by spectrophotometer .The expression levels of CATK , MMP-9,TRAP as well as OPG/RANKL/RANK mRNA and protein were detected by Real time-PCR and Western Blot .Results As compared with those in blank control group, the TRAP positive cell number and the expression levels of TRAP ,MMP-9,CATK,RANKL,RANK mRNA and protein were significantly increased in HG-DMEM induction group , however , the expression levels of OPG mRNA and protein were significantly decreased ( P <0.05).As compared with HG-DMEM induction group, after 24-hour pretreatment, naringenin could obviously decrease TRAP positive cell number and the expression levels of CATK , MMP9-, TRAP, RANKL, RANK mRNA and protein ,and could increase the expression levels of OPG mRNA and protein ,with a dose-dependent way ( P <0 .05 ) .Conclusion Naringenin can inhibit the generation and differentiation of osteoclasts ,and the action may be realized by regulating OPG/RANKL/RANK signal transduction pathway to inhibit the expressions of TRAP , MMP-9,CATK.