重组人甲状旁腺激素1-34促进小鼠间充质干细胞增殖及骨向分化.docVIP

重组人甲状旁腺激素1-34促进小鼠间充质干细胞增殖及骨向分化 易玲娴，翁土军，苏 楠，何启芬，罗凤涛，陈 林（400042 重庆，第三军医大学大坪医院野战外科研究所创伤实验室，创伤，烧伤与复合伤国家重点实验室） [摘要] 目的 研究重组人甲状旁腺激素1-34[（recombinant human parathyroid hormone1-34, rhPTH(1-34)]间断处理对小鼠骨髓间充质干细胞（bone marrow stromal cell，BMSC）增殖以及骨向分化的影响，进一步探讨rhPTH（1-34）促骨形成的机制。方法 无菌条件下分离6～8周的C3H雄性小鼠BMSC，传代培养，10 nmol/L rhPTH(1-34)间断处理或者Vehicle处理，MTT检测细胞增殖；BMSC成骨诱导后，分别在3、6、9 d进行碱性磷酸酶（alkaline phosphatase，ALP）染色和活性测定；RT-PCR检测成骨细胞功能标志基因核心结合因子α1 (core binding factor α1 ，Cbfα1)、骨桥蛋白( osteopontin， OP)、骨钙蛋白( osteocalcin， OC)的表达水平；Western blot检测p-Erk1/2,p-P38信号通路。结果 rhPTH(1-34)间断处理促进BMSC增殖；诱导9 d时与对照组比较，显著增加ALP活性[D(405)/D(562)：(0.625±0.049)vs(0.543±0.038)，(P0.05)](P0.05)；显著增加成骨功能相关基因Cbfα1(增加186.6%，P0.01),OC（增加210.5%，P0.01），OP mRNA（增加5.5%，P0.05）的表达水平；p-Erk1/2，p-P38水平均增加。结论 rhPTH(1-34)间断处理通过激活下游的p-Erk1/2和p-P38通路，促进BMSC增殖，增强其向成骨细胞分化，使成骨细胞数量和活性增加 [关键词] 甲状旁腺激素；间充质干细胞；成骨细胞 [中图法分类号] Recombinant human parathyroid hormone 1-34rhPTH（1-34） treatment increase promotes proliferation of bone marrow stromal cells in mice mouse BMSC proliferation and enhancetheir differentiateion to osteoblasts Yi Lingxian, Weng Tujun, Su Nan, He Qifen, Luo Fengtao, Chen Lin (State Key Laboratory of Trauma, Burns and Combined Injury, Trauma Center, Institute of Field Surgery Research, Third affiliatedDaping Hospital, Third Military Medical University, Chongqing ,400042, China) ［Abstract］ Objective To study the effects of recombinant human parathyroid hormone 1-34 (rhPTH( 1-34) intermittent treatment on the proliferation of bone marrow stromal cells (BMSC) and osteogenictheir differentiation to osteoblasts of BMSC, and to make a better understanding of theas well as its mechanism underlying bone formation of the anabolic effects of rhPTH(1-34) in bone. Methods: BMSC, were isolated from femurs and tibias of C3H male mice at the age of 6-8 weeks, were subcultured old. Cells were and treated with 10nM rhPTH (1-34) intermittently or with Vvehicle,. then determined it’s pProliferation of BMSC was detected by t