淫羊藿苷促进骨膜细胞增殖及其机制  

Icariin promotes the proliferation of human periosteum cells and the underlying mechanism

作　　者：李红明 高原 胡小雄 Li Hong-ming;Gao Yuan;Hu Xiao-xiong(General Department,Longhua District People’s Hospital of Shenzhen,Shenzhen 518000,Guangdong Province,China;Department of Infection,the People’s Hospital of Yichun City,Yichun 336000,Jiangxi Province,China)

机构地区：[1]深圳市龙华区人民医院全科,广东省深圳市518000 [2]宜春市人民医院感染科,江西省宜春市336000

出　　处：《中国组织工程研究》2018年第4期505-509,共5页Chinese Journal of Tissue Engineering Research

基　　金：江西省科技计划项目(20071BBG60108)~~

摘　　要：背景:骨膜细胞是成骨细胞及软骨细胞的前体细胞,虽然有研究表明骨形态发生蛋白7等可用于诱导骨膜细胞增殖,但价格较贵,不易推广。植物雌激素淫羊藿苷诱导骨膜细胞增殖给组织工程提供了新的方向。目的:观察淫羊藿苷对人骨膜细胞增殖的影响,分析其作用机制。方法:体外培养人骨膜细胞,传至第3代后,用10~3/孔的浓度种植于24孔板。首先进行细胞增殖实验:实验分为2组:实验组加入不同质量浓度淫羊藿苷(10^(-1),10^(-2),10^(-3) mg/L)及细胞培养液,对照组仅加入细胞培养液。随后探讨细胞增殖机制,实验分为2组:实验组加入雌激素受体拮抗剂ICI182780于不同质量浓度淫羊藿苷(10^(-1),10^(-2),10^(-3) mg/L)的细胞中,对照组仅加细胞培养液培养。两种实验均采用MTT法检测细胞在第1,2,3天的细胞增殖情况。采用蛋白质印迹法检测不同质量浓度的淫羊藿苷对骨膜细胞雌激受体α和雌激素β蛋白的影响。结果与结论:(1)细胞增殖:体外培养的人骨膜细胞增殖良好,干预1,2,3 d后,质量浓度10^(-1),10^(-2)及10^(-3) mg/L的淫羊藿苷均能促进细胞增殖(P<0.05);(2)细胞增殖机制分析:加入雌激素受体拮抗剂ICI182780后,质量浓度10^(-1),10^(-2),10^(-3) mg/L的淫羊藿苷对骨膜细胞均无明显的促进增殖作用(P>0.05),但可上调骨膜细胞雌激素受体α,β蛋白的表达;(3)结果证实,质量浓度10^(-1),10^(-2),10^(-3) mg/L淫羊藿苷能促进骨膜细胞增殖,其机制可能通过上调雌激素受体α,β蛋白的表达发挥作用的。BACKGROUND:Periosteal cells are precursors of osteoblasts and chondrocytes.Some studies have reported that bone morphogenetic protein-7 can be used to induce periosteal cell proliferation,but limited by the high cost.Phytoestrogen icariin-induced periosteal cell proliferation has provided a new direction for tissue engineering.OBJECTIVE:To investigate the effect of icariin on the proliferation of human periosteum cells and to analyze the underlying mechanism.METHODS:Human periosteal cells were cultured in vitro and seeded to the 24-well plate with the concentration of 103/well after third passage.Cell proliferation test:the cells were cultured in the cell culture medium(control group),or the culture medium containing different concentrations of icariin(10-1,10-2 and 10-3 mg/L).Proliferation mechanism test:the cells were cultured in the cell culture medium(control group),or the culture medium containing different concentrations of icariin(10-1,10-2 and 10-3 mg/L)plus estrogen receptor antagonist ICI182.780.The cell proliferation in each test was detected by MTT assay at 1,2 and 3 days of culture.The effects of different concentrations of icariin on the levels of estrogen receptorαandβproteins in the periosteal cells were detected by western blot assay.RESULTS AND CONCLUSION:The proliferation of human periosteum cells in vitro was successful,and icariin with the concentrations of 10-1,10-2 and 10-3 mg/L could significantly the cell proliferation(P<0.05).However,this effect was blocked after ICI182780 addition(P<0.05),and the levels of estrogen receptorαandβwere upregulated.To conclude,icariin can enhance the proliferation of periosteal cells probably by upregualting the expression of estrogen receptorαandβ.

关 键 词：淫羊藿苷 骨膜细胞 细胞增殖 诱导剂 植物雌激素 雌激素Α受体 雌激素Β受体 雌激素拮抗剂 组织构建 

分 类 号：R318[医药卫生—生物医学工程]