淫羊藿甙对体外钛微粒诱导破骨细胞骨吸收功能的影响  

Effect of Icariine on Osteoclast Bone Resorption Induced by Titanium Particle in vitro

作　　者：张怡元[1] 林煜[1] 冯尔宥[1] 肖莉莉[1] 陈嵘[1] 王武炼[1] 林丽琼[1] 

机构地区：[1]厦门大学附属福州市第二医院关节外科,福州350007

出　　处：《中国中医骨伤科杂志》2013年第10期4-7,共4页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：福建省青年人才基金(2008F3121);福建省卫生厅青年人才基金(2007-2-37);福州市科技局基金(2009-G-104)

摘　　要：目的:探讨淫羊藿甙对钛微粒诱导的体外培养的破骨细胞骨吸收功能的影响。方法:用巨噬细胞集落刺激因子(macrophage colony stimulating factor,M-CSF)和核因子-κB受体活化因子配体(receptor activator of NF-kB ligan,RANKL)诱导破骨细胞前体RAW 264.7细胞为破骨细胞,MTT法检测不同浓度淫羊藿甙对RAW 264.7细胞增殖的影响,得出最佳浓度。实验分为正常培养基的钛微粒处理组、含淫羊藿甙的钛微粒处理组和空白对照组,48h后收集细胞,采用ELISA法检测培养液中细胞核因子k B活化因子受体的浓度(receptor activator of NF-kB,RANK),用实时荧光定量PCR法检测抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase,TRAP)、基质外金属蛋白酶-9(matrix metalloproteinase 9,MMP-9)、碳酸酐酶Ⅱ(carbonic anhydraseⅡ,CAⅡ)、组织蛋白酶K(Cathepsin K,CtsK)mRNA的表达;Western Blot法检测TRAP、RANK、CtsK蛋白的表达。设置同样的实验组与骨磨片共培养,甲苯胺蓝染色计算骨吸收陷窝的数目和面积。结果:淫羊藿甙可明显下调钛微粒诱导的RAW 264.7细胞和细胞液中RANK的表达,TRAP、MMP-9、CAⅡ、CtsKmRNA和蛋白的表达,并减少骨磨片上骨吸收陷窝的数目和面积。结论:淫羊藿甙可抑制钛颗粒体外诱导破骨细胞的骨吸收,有望成为防治人工关节无菌性松动的药物。Objective：To study the effect of icariine on osteoclast bone resorption induced by titanium particle in vitro. Methods：maerophage colony stimulating factor （M-CSF） and receptor activator of NF-kB ligand （RANKL） induced osteoclast precurs RAW 264.7 cells to osteoclast. The best in vitro icariine serum concentration for promoting proliferation of osteo- clast was obtained by MTT assay. RAW 264.7 cells were treated with titanium particle with or without icariine for 48h. The expression of receptor activator of NF-kB （RANK） was measured by ELISA. The expression of TRAP, MMP-9, CA Ⅱ and CtsK mRNA in RAW 264.7 cells were detected by Real time PCR. The expression of TRAP, RANK and CtsK protein was measured by Western Blot. Toluidine blue staining was employed to calculate the number and area of resorption pits in gound bone section. Results：Icariine obviously decreased the expressions of RANK, TRAP, MMP-9, CAⅡ and the number and area of resorption pits in gound-bone section. Conclusion：Icariine could inhibit osteoclast bone resorption ability and may be a promising drug for treating aseptic loosening.

关 键 词：淫羊藿甙 钛微粒 RAW264 7 骨吸收 假体松动 

分 类 号：R-33[医药卫生]