马钱子碱对多发性骨髓瘤中成骨细胞及破骨细胞代谢的影响

摘要

This study was aimed to explore the influence of brucine on the early differentiation of osteoblasts and the metabolic pathway of osteoclast in multiple myeloma (MM) and to compare the effects of brucine and bontezomib on MM.The half inhibitory concentration( IC50 ) of brucine and bortezomib on MM cell line U266 was determined by MTT method; the mRNA levels of alkaline phosphatase(ALP), osteocalcine(OC), osteoprotegerin(OPG) and osteoprotegrin ligand(RANKL) were detected by RT-PCR after the supernatant of cultured U266 cells was added into the culture system for inducing the differentiation of osteoblast line MC3T3-E1 and culturing.The results showed that the IC50 of bortezomib and brucine on U266 cells for 48 hours were 22.4 nmol/L and 0.16 mg/ml respectively.As compared with osteoblasts treated by supernatant of cultured MM cells alone, the mRNA levels of ALP, OC and OPG in osteoblasts treated by brucine combined with supernatant of cultured MM cells were enhanced (p ＜ 0.05 ), while the RANKL mRNA level was lowed (p ＜ 0.05 ), moreover the enhanced and lowed degree also was large (p ＜ 0.05 ).It is concluded that the influence of brucine on metabolism of osteoblasts and osteoclasts in MM may be realized through the regulation of osteoclasts by osteoblasts.The therapeutic efficacy of brucine on MM is superior to bortezomib.%本研究旨在探讨体外马钱子碱对多发性骨髓瘤(MM)中成骨细胞早期分化和破骨细胞代谢途径的影响,同时比较马钱子碱与硼替佐米在体外对多发性骨髓瘤的影响.用MTT法检测硼替佐米与马钱子碱对多发性骨髓瘤细胞株U266的半数抑制浓度(IC(50));将多发性骨髓瘤细胞株U266的上清液加入到成骨细胞MC3T3-E1诱导分化体系中培养后,用RT-PCR方法测定碱性磷酸酶(ALP)、骨钙蛋白(OC)、骨保护蛋白(OPG)及骨保护蛋白配体(RANKL)的mRNA水平.结果表明,硼替佐米作用于多发性骨髓瘤细胞株U266 48小时后半数抑制浓度(IC(50))为22.4 nmol/L,马钱子碱为0.16 mg/ml;经马钱子碱及多发性骨髓瘤细胞上清液共同干预组的成骨细胞中ALP、OC及OPG的mRNA水平高于只经多发性骨髓瘤细胞上清液干预组的成骨细胞的表达水平(p<0.05),而RANKL的mRNA水平则降低(p<0.05),且它们增高或降低的程度大于硼替佐米作用组(p<0.05).结论:马钱子碱对多发性骨髓瘤中骨代谢机制的影响可能通过成骨细胞对破骨细胞的调节而发挥作用.实验证实,马钱子碱时多发性骨髓瘤的治疗效果优于硼替佐米.