促黑素对小鼠成骨细胞OPG/RANKLmRNA表达的影响

Effect of α-melanocyte stimulating hormone on the expressions of OPG and RANKL mRNA in mouse osteoblasts

目的探讨原代培养的小鼠成骨细胞黑皮质素受体(MCR)表达的种类及促黑素(α-MSH)对其骨保护素(OPG)和核因子-κB受体活化因子配体(RANKL)mRNA表达的影响。方法酶消化法分离3 d内新生C57BL/6小鼠原代成骨细胞,培养液中分别加入10-7、10-8和10-9mol/L的α-MSH培养12 h,作为A、B、C组;培养液中加入10-7mol/Lα-MSH,分别培养6、12、24 h,作为D、E、F组。对照组正常培养。采用实时定量PCR方法分析α-MSH对成骨细胞OPG/RANKL mRNA表达的影响。利用RT-PCR法测定原代成骨细胞MCR的表达。结果小鼠原代成骨细胞表达MC1R、MC2R、MC4R和MC5R;与对照组相比,α-MSH促进OPG/RANKL mRNA的表达(P<0.05)。结论α-MSH可能通过MCR系统促进小鼠原代成骨细胞OPG/RANKL mRNA的表达。

Objective To investigate the types of melanocortin receptor （MCR） of the primary osteoblasts, and the effect of α-melanocyte stimulating hormone （ct-MSH） on the expressions of osteoprotegerin （OPG） and receptor activator of nuclear factor-kappa B ligand （RANKL） mRNA in mouse osteoblasts. Methods Calvarial osteoblasts were separated and cultured in vitro by the mechanical separation and enzyme digestion method. The primary osteoblasts were divided into groups A, B and C, which were treated with 10^-7, 10^-8 and 10^-9 mol/L et-MSH respectively for 12 h; the primary osteoblasts were divided into groups D, E and F, which were treated with 10^-7 mol/L α-MSH for 6, 12 and 24 h respectively. Control group was set up at the same time. The expressions of OPG and RANKL mRNA were detected by Real-Time PCR. MCR subtypes in the primary osteoblasts were measured by RT-PCR. Results MC1R, MC2R, MC4R and MC5R were expressed in primary cultured osteoblasts. The expressions of OPG and RANKL mRNA significantly increased after incubation with α-MSH （P 〈 0.05）. Conclusion α-MSH might promote the expressions of OPG and RANKL mRNA in the primary osteoblasts through MCR system.