葛根素联合国产多孔钽对人成骨细胞COL-I、OC、OPN表达和细胞增殖的影响  

The influence of puerarin combined with domestic porous tantalum on the expression of COL-I,OC,OPN and cell proliferation of osteoblasts

作　　者：刘永庆 李琪佳 甘洪全 王茜 张大鹏 王志强 

机构地区：[1]华北理工大学,河北唐山063000 [2]华北理工大学医学中心实验室,河北唐山063000 [3]华北理工大学附属医院骨科,河北唐山063000 [4]华北理工大学基础医学院,河北唐山63000

出　　处：《中国骨质疏松杂志》2017年第2期236-243,共8页Chinese Journal of Osteoporosis

基　　金：国家科技部科技支撑课题(2012BAE06B03);河北省科技支撑资助项目(16277776D);河北省医学科学研究重点课题计划资助项目(20160225);河北省卫生计生委临床医学优秀人才研究项目(361036);华北理工大学博士科研启动基金资助项目

摘　　要：目的研究探索葛根素联合国产多孔钽支架材料对人成骨细胞MG-63 COL-I、OC、OPN表达和细胞增殖的影响。方法实验分为:葛根素组:不同浓度的葛根素与成骨细胞共培养(CCK-8法筛选出葛根素作用最佳剂量浓度组);多孔钽组:国产多孔钽与成骨细胞共培养;葛根素多孔钽组:葛根素、国产多孔钽与成骨细胞共培养;空白组:成骨细胞培养。扫描电镜观察多孔钽及葛根素多孔钽-成骨细胞复合物表面形态;CCK-8法检测成骨细胞的生长增殖状态;实时荧光定量PCR法(RT-PCR)和免疫细胞化学染色法检测成骨细胞COL-I、OC、OPN mRNA和蛋白的表达。结果葛根素和多孔钽组较空白组均可促进成骨细胞的增殖(P<0.05);葛根素多孔钽组较其它各组细胞的增殖活性最显著(P<0.05)。RT-PCR法和免疫细胞化学染色法结果显示,葛根素和国产多孔钽对成骨细胞COL-I、OCmRNA和蛋白表达均有影响(P<0.05),但对OPN mRNA和蛋白无显著影响(P>0.05)。结论葛根素和国产多孔钽支架具有协同影响成骨细胞COL-I、OC、OPN mRNA和蛋白的表达,以及促进成骨细胞的增殖。Objective To explore the influence of Puerarin combined with domestic porous tantalum on the expression of COL-I,OC and OPN and cell proliferation of osteoblasts.Methods There were four experimental groups including Puerarin group：different concentrations Puerarin co-cultured with osteoblasts（CCK-8 method was used to screen the optimal concentration dose group）;Porous tantalum group：domestic porous tantalum co-cultured with osteoblasts;Puerarin porous tantalum group：Puerarin and domestic porous tantalum co-cultured with osteoblasts;Blank group：osteoblast culture.Scanning electron microscopy was used to observe the surface morphology of porous tantalum and puerarin porous tantalum osteoblasts.CCK-8 method was used to detect the growth and proliferation of osteoblasts.Real-time fluorescence quantitative PCR assay（RT-PCR） and immunocytochemistry staining were used to detect the expression of COL-I,OC,mRNA OPN and protein in osteoblasts.Results Compared with the blank group,the proliferation of osteoblasts could be promoted by puerarin and porous tantalum（P 〈0.05）.Compared with other groups,the proliferation activity of the puerarin porous tantalum group was the most significant（P 〈 0.05）.RT-PCR and immunocytochemistry staining showed that Puerarin and domestic porous tantalum have effect on the expression of COL-I,OCmRNA and protein in osteoblasts（P 〈 0.05）,but there was no significant effect on OPN mRNA and protein（P 〉 0.05）.Conclusion Puerarin and domestic porous tantalum stent have synergistic effect the expression of COL-I,OC mRNA OPN and protein,as well as promoting the proliferation of osteoblasts.

关 键 词：中医中药 葛根素 多孔钽 I型胶原 骨钙素 骨桥蛋白 

分 类 号：R684[医药卫生—骨科学]