淫羊藿苷体外诱导骨膜细胞增殖及分化

Proliferation and differentiation of periosteum cells induced by icariin

背景:淫羊藿苷用于促进细胞增殖有广阔的运用前景。骨膜细胞有特定的分化方向不确定性,同时也容易被诱导增殖,常见的诱导物质包括超声、氧等物理方法或者骨形态发生蛋白7等诱导因子,目前骨组织工程的研究已经有骨膜细胞运用的相关报道。但是淫羊藿苷诱导骨膜细胞增殖相关的报道少见,淫羊藿苷诱导骨膜细胞后的分化方向目前尚未明确。目的:探讨淫羊藿苷对人骨膜细胞增殖及分化的影响,为淫羊藿苷运用于骨组织工程提供理论依据。方法:手术获得人骨膜,体外分离获得原代细胞,培养扩增后,种植培养骨膜细胞于24孔板中。分别用0.001,0.01及0.1 mg/L的淫羊藿苷及50μg/L的骨形态发生蛋白7诱导骨膜细胞增殖,MTT法检测各组骨膜细胞的吸光度值。于1,3,5和7 d检测骨膜细胞碱性磷酸酶和钙结节的数量表达情况,RT-PCR法检测骨钙素、骨桥蛋白和Runx2的mR NA含量。结果与结论:(1)骨膜细胞在加入淫羊藿苷后增殖良好,不同质量浓度的淫羊藿苷组及阳性对照组均有促进骨膜细胞增殖的作用(P<0.05);(2)加入了淫羊藿苷的骨膜细胞与对照组相比能产生更多的碱性磷酸酶和钙结节(P<0.05);(3)0.1 mg/L的淫羊藿苷显著上调骨钙素、骨桥蛋白和Runx2 mR NA的表达(P<0.01);(4)结果表明,淫羊藿苷有促进骨膜细胞增殖和分化为成骨细胞的作用,可作为诱导剂用于骨组织工程中种子细胞的制备。

BACKGROUND:Icariin has a broad prospect for promoting cell proliferation.Differentiation direction of periosteal cells is uncertain,but the cells are easy to be induced by ultrasound,oxygen or bone morphogenetic protein 7(BMP7).Periosteal cells have been applied in bone tissue engineering;however,icariin effects on the proliferation and differentiation of periosteal cells is little reported.OBJECTIVE:To investigate the effect of icariin on the proliferation and differentiation of human periosteal cells,thus providing theoretical basis for icariin applied in bone tissue engineering.METHODS:The human periosteum was obtained and the primary cells were isolated in vitro.After culture and expansion,periosteal cells were cultured in 24-well plates,and induced by 0.001,0.01 and 0.1 mg/L icariin and 50μg/L BMP7,respectively.The corresponding avsorbance values of different groups were detected.The levels of alkaline phosphatase and calcium nodules in periosteal cells were measured at 1,3,5 and 7 days,and the mRNA levels of osteocalcin,osteopontin and Runx2 were detected at 3,5 and 7 days.RESULTS AND CONCLUSION:The periosteal cells proliferated well after induction with icariin,and could proliferate well in different concentrations of icariin and the positive control group(P<0.05).Compared with the control group,the periosteal cells induced by icariin were able to produce more alkaline phosphatase and calcium nodules(P<0.05).The mRNA expression of osteocalcin,osteopontin and Runx2 in periosteal cells could be up-regulated by icariin(P<0.05).These findings imply that icariin can promote proliferation and differentiate of periosteal cells into osteoblasts,and it can be used as an inducer for the preparation of seed cells in bone tissue engineering.