马钱子碱对多发性骨髓瘤骨病骨代谢影响的体外研究

Effects of brucine on metabolism in multiple myeloma

目的探讨马钱子碱体外对多发性骨髓瘤（MM）骨病骨代谢的影响，同时与硼替佐米比较体外对MM骨病的疗效。方法应用马钱子碱和硼替佐米作用于MM细胞株U266，MTT法检测硼替佐米与马钱子碱对MM细胞株U266的半数抑制浓度（IC,o），将成骨细胞株MC3T3-E1加入有马钱子碱或硼替佐米作用的MM细胞株U266上清中培养，分为空白对照组、U266上清干预组、硼替佐米作用的U226细胞上清干预组、马钱子碱作用的U266细胞上清干预组，以RT—PCR法测定各组MC3T3-E1细胞碱性磷酸酶（ALP）、骨钙素（OC）、骨保护素（OPG）及NF—KB受体活化因子的配体（RANKL）的mRNA水平。结果硼替佐米作用于MM细胞株U26648h的IC50为22．4nmol／L，马钱子碱为0．16mg／ml；经马钱子碱作用的U266细胞上清液干预组的MC3T3。E1细胞中的ALP、OC及OPG的mRNA水平高于只经U266细胞上清液干预组（P〈0．05），而RANKL的mRNA水平则降低（P〈0．05），且4个基因mRNA水平增高或降低的程度大于经硼替佐米作用的U266细胞上清液干预组的MC3T3-E1细胞表达水平（P〈0．05）。结论马钱子碱对MM骨病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；体外马钱子碱对MM骨病骨代谢的影响效果大于硼替佐米。

Objective To explore the effects of brucine on multiple myeloma （MM） and to compare the effects between brucine and bontezomib on MM. Methods MTF method was used to determine the median inhibitory concentratiom （ICso） of brucine and bortezomib on the MM cell line U266. The supernatant of cultured U266 cell line was added to the culture system for inducing the differentiation of osteoblast cell line MC3T3-EI. After aseptic assay, RT-PCR was used to determine the RNA levels of alkaline phosphatase （ALP）, osteocalcine （OC）, osteoprotegcrin （OPG） and recepter activator of NF-KB ligand （RANKL）. Results IC50 of bortezomib on U266 cell line for 48 h was 22.4 nmol/L, and that of strychnine was 0.16 nmol/L. The mRNA levels of ALP, OC and OPG in osteoblast co-intervened by brucine combined with the supernatant of MM cells were higher than those in supernatant of U266 cells, while the level of RANKL mRNA was lower （P 〈0.05）. The degree of increasing or reducing was greater than the level of control group intervened only by bortezomib （P 〈0.05）. Conclusion The therapeutic effects of brucine on MM might be carried out through the regulation of osteoclast by osteoblast, and the experiment confirmed that the therapeutic effect of brucine on MM was superior to that of bortezomib.