硫酸钙对成骨样MG-63细胞增殖及骨保护素/核激活因子受体配体/核激活因子受体系统的影响研究  

Effects of calcium sulfate on proliferation of osteoblast-like MG-63 cells and osteoprotegerin/receptor activator of NF-κB ligand/receptor activator of NF-κB system

作　　者：李正茂 陈斌 崔志 谭文甫 何敏 Li Zhengmao;Chen Bin;Cui Zhi;Tan Wenfu;He Min(Department of Traumatic Orthopedics,The Second Affiliated Hospital,University of South China,Hengyang 421001,Hunan,China)

机构地区：[1]南华大学附属第二医院创伤骨科,湖南省衡阳市421001

出　　处：《中华创伤骨科杂志》2021年第8期707-710,共4页Chinese Journal of Orthopaedic Trauma

基　　金：湖南省卫生健康委科研项目(202104072014);衡阳市科技局指导性科研项目(163)。

摘　　要：目的探讨硫酸钙对成骨样MG-63细胞增殖情况及骨保护素/核激活因子受体配体/核激活因子受体(OPG/RANKL/RANK)系统的影响。方法制作硫酸钙浸提液,将含有硫酸钙浸提液的细胞培养基设为硫酸钙组,不含硫酸钙的普通细胞培养基设为空白对照组,分别培养成骨样MG-63细胞24 h。观察两组细胞的生长情况,通过细胞增殖实验(CCK-8)检测细胞的增殖活性,并检测OPG/RANKL的mRNA和蛋白表达水平。结果硫酸钙组和空白对照组成骨样MG-63细胞的生长情况均较好。在CCK-8增殖试验中,硫酸钙组与空白对照组的吸光度值分别为0.997±0.008、0.640±0.003,差异有统计学意义(P<0.001)。硫酸钙与空白对照组OPG的mRNA相对表达量分别为2.834±0.176、1.005±0.102;RANKL的mRNA相对表达量分别为0.355±0.035、1.002±0.068,差异均有统计学意义(P<0.001)。蛋白质免疫印记结果显示:与空白对照组相比,硫酸钙组中的硫酸钙能促进成骨样MG-63细胞OPG的蛋白表达,并抑制RANKL的蛋白表达。结论硫酸钙能促进成骨细胞样MG-63细胞的增殖与活性,并可调控OPG/RANKL/RANK系统,具有一定的促骨形成作用。Objective To investigate the effects of calcium sulfate on the proliferation of osteoblast-like MG-63 cells and the osteoprotegerin/receptor activator of NF-κB ligand/receptor activator of NF-κB(OPG/RANKL/RANK)system.Methods The extract of calcium sulfate was prepared.The osteoblast-like MG-63 cells were cultured for 24 hours in the medium containing calcium sulfate(the calcium sulfate group)and in the normal medium without calcium sulfate(the blank group),respectively.The growth of osteoblast-like MG-63 cells was observed and their proliferation detected by CCK-8.The mRNA and protein expression levels of OPG/RANKL were detected.Results The growth of osteoblast-like MG-63 cells was fine in both groups.The CCK-8 test showed that the absorbance value at 24 h was 0.997±0.008 for the calcium sulfate group,significantly higher than that for the blank group(0.640±0.003)(P<0.001).Respectively,the mRNA expression levels of OPG were 2.834±0.176 and 1.005±0.102 and the mRNA expression levels of RANKL 0.355±0.035 and 1.002±0.068 for the calcium sulfate group and the blank control group,showing statistically significant differences(P<0.001).The results of Western blot showed that compared with the blank control group,the protein expression of OPG in osteoblast-like MG-63 cells was promoted but the protein expression of RANKL inhibited in the calcium sulfate group.Conclusion Calcium sulfate may have a positive effect on bone formation,because it can promote the proliferation and activity of osteoblast-like MG-63 cells and regulate the OPG/RANKL/RANK system.

关 键 词：硫酸钙 细胞增殖 骨生成 成骨样MG-63细胞 

分 类 号：R318.08[医药卫生—生物医学工程]