淫羊藿甙对高浓度葡萄糖抑制MC3T3-E1细胞成骨分化的保护作用  

Protective Effect of Icariin on the Osteoblasts Differentiation of MC3T3-E1 Cells Against Inhibition Effect of High Concentration of Glucose

作　　者：吴宁 康乐[1] 革军 权冬 陈臣 庞炜[1] WU Ning;KANG Le;GE Jun;QUAN Dong;CHEN Chen;PANG Wei(Department of Orthopaedics,No 323 Hospital of the PLA 1,Xi'an 710054,China;Department of Orthopaedics,No 518 Hospital of the PLA 2,Xi'an 710043,China)

机构地区：[1]解放军第三二三医院骨科,西安710054 [2]解放军第五一八医院骨科,西安710043

出　　处：《科学技术与工程》2018年第33期13-18,共6页Science Technology and Engineering

基　　金：陕西省重点研发计划(2017SF-223)资助

摘　　要：为了探讨淫羊藿甙对高浓度葡萄糖抑制MC3T3-E1细胞成骨分化是否具有保护左右,采用含10 mmol/Lβ-磷酸甘油、50 mg/L维生素C以及10^(-8)mol/L地塞米松的α-MEM培养基诱导MC3T3-E1细胞成骨分化。根据培养基葡糖糖含量不同分别设置对照组(5. 5 mol/L葡萄糖)、高糖组(22 mol/L葡萄糖)和药物干预组(22 mol/L葡萄糖)。药物干预组各组细胞换液时分别加入0. 1μmol/L、1. 0μmol/L和10μmol/L的淫羊藿甙。通过茜素红染色、钙含量检测方法检测诱导培养25 d后细胞外钙含量,使用Real time PCR检测诱导培养7 d后成骨标志分子表达,观察各组MC3T3-E1细胞的成骨分化情况。使用活性氧检测试剂盒检测诱导培养7 d后的各组MC3T3-E1细胞内自由氧(reactive oxygen species,ROS)生成水平。结果表明:22mmol/L葡萄糖组细胞钙含量最低,成骨标志分子ALP、Osteorix及Runx2表达降低,细胞内自由氧生成水平升高,MC3T3-E1细胞成骨分化受到抑制。使用不同浓度淫羊藿甙干预后,细胞钙含量升高,成骨标志分子ALP、Osteorix及Runx2表达升高,细胞内自由氧生成水平降低,MC3T3-E1细胞成骨分化抑制作用呈现浓度依赖性的减弱。可见淫羊藿甙能够通过降低细胞内自由氧水平,减弱高浓度葡萄糖对成骨细胞分化的抑制作用。In order to investigate whether Icariin had protective effect on the inhibition of MC3T3-E1 cell osteogenic differentiation by high concentration of glucose in vitro.MC3T3-E1 cells were induced into osteogenic differentiation usingα-MEM calcification medium containing 10mmol/Lβ-glycerophosphate,50 mg/L vitamin C and 10-8 mol/L dexamethasone.MC3T3-E1 cells were divided into five groups:negative control(NC,cells cultured in osteogenic induction medium with 5.5 mmol/L glucose),high glucose(HG,cells cultured in osteogenic induction medium with 22 mmol/L glucose)and Icariin intervention(ICA,cells cultured in osteogenic induction medium with 22 mmol/L glucose,in the presence of 0.1μmol/L,1.0μmol/L and 10μmol/L icariin,respectively).The extracellular calcium deposition of MC3T3-E1 cells was observed by alizarin red staining,calcium content detection in 25 days of cultures.The mRNA expression of osteoblast markers(ALP,Osteorix and Runx2)were detect by real time PCR in 7 days of cultures.Intracellular reactive oxygen species(ROS)generation level of each group were detected by Reactive Oxygen Species Assay Kit in 7 days of cultures.The results show that 22 mmol/L glucose suppressed the osteoblast differentiation process of MC3T3-E1 cell and the intracellular ROS generation increased,in which,the calcium deposition and osteoblast markers were reduced.Icariin could significantly reverse the suppression effect of high glucose and decrease of above-mentioned osteoblast differentiation markers and diminish the intracellular ROS generation.It is concluded that Icariin prevents the damage of osteoblast differentiation under high concentration glucose by decreasing ROS generation.

关 键 词：糖尿病性骨质疏松症 淫羊藿甙 ROS 成骨细胞分化 

分 类 号：Q591.9[生物学—生物化学] R681[医药卫生—骨科学]