淫羊藿苷对人成骨样细胞成骨分化及OPG/RANKL表达的影响

Effects of icariin on the differentiation of osteoblast and regulation of the OPG-RANKL system in MG-63 cells

目的探讨淫羊藿苷对人成骨样细胞(MG-63)成骨分化及OPG/RANKL表达的影响。方法用1 nmol/L、10 nmol/L、100 nmol/L、1μmol/L、10μmol/L 5种浓度的淫羊藿苷对MG-63进行干预,并同时进行成骨诱导,RT-PCR在第3天检测其细胞增殖基因MYC、CDK7表达量,在第6天检测成骨分化标志基因RUNX2、COL1A1及OPG、RANKL的表达量。结果对成骨分化相关基因,10 nmol、1μmol干预的MG-63 RUNX2基因表达量较1 nmol及10μmol的高(P<0.05),与100 nmol及空白对照组没有明显差别(P>0.05),10μmol的RUNX2蛋白表达量较其余各组更高;COL1A1的基因表达呈剂量依赖型,随浓度的增加而升高,10μmol的表达量较其余各组有明显升高(P<0.05),其蛋白表达也更多;RANKL的表达量极低,且在各组之间均无明显差异(P>0.05);OPG的表达在1μmol浓度时较100 nmol、10μmol明显增高(P<0.05),与1 nmol、10 nmol及空白组没有统计学差异(P>0.05);与细胞增殖相关的MYC的表达各组之间均没有差异(P>0.05),而CDK7的表达均较空白组降低(P<0.05)。结论 10μmol/L的淫羊藿苷能够明显促进MG-63细胞的成骨分化,但并不通过影响OPG/RANKL起效。

Objective To explore the effects of icariin on the differentiation of osteoblast and regulation of the OPG-RANKL system in MG-63 Cells. Methods The MG-63 cells were cultured in icariin at 1 nmol / L,10 nmol / L,100 nmol / L,1 μmol / L,10μmol /L,and induced to osteoblast at the same time. The mRNA and protein expression of RUNX2,COL1A1,OPG,and RANKL at6 th day,and mRNA expression of MYC,CDK7 at 3rd day,was analyzed by quantitative reverse transcription polymerase chain reaction（ RT-q PCR） and western-blot,respectively. Results Compared with the control group,the RUNX2,OPG,RANKL,MYC,expression showed no difference with icariin stimulated（ P〉0. 05）,but the CDK7 was reduced singnificantly（ P〈0. 05）.Icariin at 10 umol produced a higher COL1A1（ P〈0. 05） which was the marker gene of osteoblast. Conclusion Icariin at 10 umol promot-edosteoblast differentiation without affecting OPG-RANKL system.