葛根素通过NO信号通路促进体外培养大鼠骨髓间充质细胞成骨性分化  

Puerarin Promoted Bone Marrow Mesenchymal Cells Osteogenesis Differentiation of Rats Through NO Signaling Pathway

作　　者：宋明甲 文益民 柴晓亮[1] 周建[2] 吴晓燕[1] SONG Mingjia;WEN Yimin;CHAI Xiaoliang;ZHOU Jian;WU Xiaoyan(Spinal Surgery,Joint Logistics Support Force 940 Hospital,CPLA,,Lanzhou 730050,China;Institute of Orthopaedics,Joint Logistics Support Force 940 Hospital,CPLA,Lanzhou 730050,China)

机构地区：[1]中国人民解放军联勤保障部队九四〇医院脊柱外科,兰州730050 [2]中国人民解放军联勤保障部队九四〇医院骨科研究所,兰州730050

出　　处：《中国现代应用药学》2019年第17期2124-2130,共7页Chinese Journal of Modern Applied Pharmacy

基　　金：甘肃省自然科学基金资助项目(1506RJZA306)

摘　　要：目的观察葛根素促进体外培养大鼠骨髓间充质细胞成骨性分化过程中是否激活NO信号通路。方法一氧化氮合成酶阻断剂(nitric oxide synthase inhibitors,LNMA)预处理大鼠骨髓间充质细胞12 h后,使用葛根素处理骨髓间充质细胞不同时间,观察其对骨钙素(osteocalcin,OC)含量,碱性磷酸酶(alkaline phosphatase,ALP)活性和钙化结节染色,Runx-2、OSX、BMP-2和Collagen-1 mRNA表达水平的影响,以及NO和3’-5’-环鸟苷一磷酸(cGMP)含量的影响。结果葛根素对骨髓间充质细胞增殖活性存在浓度的依赖性,其中1×10^-6 mol·L^-1促进其增殖活性最高,并显著提高了该细胞中ALP活性。预先使用LNMA处理大鼠骨髓间充质细胞后,葛根素提高该细胞中ALP活性、OC含量、钙化能力和Runx-2、OSX、BMP-2和Collagen-1 mRNA的表达水平作用均受到抑制,以及葛根素提高NO和cGMP含量能力受到抑制。结论葛根素能有效促进体外培养大鼠骨髓间充质细胞成熟与矿化,并需要NO信号通路的参与。OBJECTIVE To observe whether puerarin promotes rat bone marrow mesenchymal cells osteogenesis differentiation through NO signaling pathway in vitro.METHODS The rat bone marrow mesenchymal cells were pretreated with nitric oxide synthase inhibitors(LNMA)12 h,to observe the presence of puerarin on osteocalcin(OC),alkaline phosphatase(ALP)and calcification nodules staining,PCR detected the expression levels of osteogenic genes Runx-2,OSX,BMP-2,and Collagen-1 mRNA and determined NO and 3’-5’-Cycloguanosine monophosphoric acid(3’-5’-cyclic-GMP,cGMP)after different processing times.RESULTS After treatment of rat bone marrow mesenchymal cells with different concentrations of puerarin,the group of 1×10^-6 mol·L^-1 significantly increased the activity of ALP in rat bone marrow mesenchymal cells and pre-treated with LNMA.After the rat bone marrow mesenchymal cells.The enhancement of ALP activity,OC content,calcification ability,and expression levels of Runx-2,OSX,BMP-2,and Collagen-1 mRNA in rat bone marrow mesenchymal cells were inhibited.It was further verified that puerarin could significantly increase NO and cGMP content,but the pretreatment with LNMA was inhibited.CONCLUSION Puerarin can effectively promote the maturation and mineralization of rat bone marrow mesenchymal cells in vitro,and NO signaling pathway is required to participate.

关 键 词：葛根素 骨髓间充质细胞 NO/cGMP信号通路 

分 类 号：R965.2[医药卫生—药理学]