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川续断皂苷Ⅵ诱导大鼠骨髓间充质干细胞向成骨细胞方向分化的研究
引用本文:武密山,赵素芝,任立中,王茹,白霞,韩红伟,李彬.川续断皂苷Ⅵ诱导大鼠骨髓间充质干细胞向成骨细胞方向分化的研究[J].中国药理学通报,2012,28(2):222-226.
作者姓名:武密山  赵素芝  任立中  王茹  白霞  韩红伟  李彬
作者单位:1. 河北医科大学中医学院方剂学教研室,河北,石家庄,050091
2. 石家庄市桥东区医院,河北,石家庄,050041
3. 河北医科大学第一医院骨科,河北,石家庄,050031
4. 河北医科大学生物化学教研室,河北,石家庄,050017
基金项目:国家自然科学基金资助项目(No 81073074,30472200);河北省引进留学人员经费资助项目(No 200828)
摘    要:目的探讨川续断皂苷Ⅵ(akebia saponin D)对大鼠骨髓间充质干细胞(MSCs)定向诱导分化为成骨细胞的影响。方法用密度梯度离心结合贴壁培养法分离纯化大鼠骨髓MSCs,选择合适的培养基,传代扩增。采用不同浓度的川续断皂苷Ⅵ对大鼠MSCs定向诱导分化,MTT法观察各组对MSCs的增殖作用并绘制细胞生长曲线;观察MSCs诱导分化过程中成骨细胞内碱性磷酸酶(alkaliphosphatase,ALP)活性;采用ELISA法检测MSCs诱导分化过程中骨钙素(Osteo-calcin,OC)的含量;RT-PCR方法检测MSCs诱导分化过程中核心结合因子α-1(Cbfα1)mRNA表达。结果细胞生长曲线显示各组MSCs数量均随时间延长而增加,中、高浓度的川续断皂苷Ⅵ能明显提高MSCs分化为成骨细胞的ALP活性和骨钙素的含量,且使Cbfα1 mRNA的表达升高。结论川续断皂苷Ⅵ具有促进大鼠体外MSCs向成骨细胞增殖和分化的作用,这一作用可能与其升高Cbfα1 mRNA的表达有关。

关 键 词:川续断皂苷Ⅵ  骨髓间充质干细胞  成骨细胞  细胞增殖  细胞分化  碱性磷酸酶  核心结合因子α-1

Experimental study of akebia saponin D on the differentiation of rat bone marrow derived mesenchymal stem cells to osteoblasts in vitro via induction
WU Mi-shan,ZHAO Su-zhi,REN Li-zhong,WANG Ru,BAI Xia,HAN Hong-wei,LI Bin.Experimental study of akebia saponin D on the differentiation of rat bone marrow derived mesenchymal stem cells to osteoblasts in vitro via induction[J].Chinese Pharmacological Bulletin,2012,28(2):222-226.
Authors:WU Mi-shan  ZHAO Su-zhi  REN Li-zhong  WANG Ru  BAI Xia  HAN Hong-wei  LI Bin
Institution:1.Dept of Formulaology,Traditional Chinese Medicine College,Hebei Medical University,Shijiazhuang 050091,China; 2.Qiaodong District Hospital of Shijiazhuang,Shijiazhuang 050041,China; 3.Dept of Orthopaedic,the First Hospital of Hebei Medical University,Shijiazhuang 050031,China; 4.Dept of Biochemistry,Hebei Medical University,Shijiazhuang 050017,China)
Abstract:Aim To investigate the role of akebia saponin D on the differentiation of rat bone marrow derived mesenchymal stem cells(MSCs)to osteoblasts in vitro via induction.Methods SD rats were sacrificed,the marrow MSCs were separated and purified by density gradient centrifugation and by adhering to the culture plastic,suitable culture medium was selected,after the in vitro secondary culture and expansion.The bone marrow derived mesenchymal stem cells were cultured in the culture media with varied akebia saponin D of different concentrations.MTT was used to observe the proliferation,and the growth curve was obtained.The alkaline phosphata(ALP)activity changes were observed in MSCs cells during differentiation being induced.Osteocalcin content of MSCs cells during differentiation was detected by ELISA.Core-binding factor alpha 1(Cbfα1) mRNA expressions of MSCs cells during differentiation were detected by RT-PCR.Results The growth curve showed that the MSCs quantities of each group had increased accompanied with time.Higher concentration and middle concentration of akebia saponin D with inductor significantly enhanced the ALP activity and osteocalcin content of MSCs induced to osteoblasts,and promoted the expression of Cbfα1 mRNA.Conclusion Akebia saponin D can enhance significantly the proliferation and differentiation of rat bone marrow derived mesenchymal stem cells to osteoblasts in vitro via induction,which may be mediated by increasing the expression of Cbfα1 mRNA.
Keywords:akebia saponin D  mesonchymal stem cells  osteoblast  cell proliferation  cell differentiation  alkaline phosphata  core-binding factor alpha 1
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