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复方扶芳藤合剂含药血清促进骨髓间充质干细胞增殖的作用机制

吴玲玲, 陈继冰, 蒋鹏, 等. 复方扶芳藤合剂含药血清促进骨髓间充质干细胞增殖的作用机制[J]. 器官移植, 2022, 13(3): 363-370. doi: 10.3969/j.issn.1674-7445.2022.03.013
引用本文: 吴玲玲, 陈继冰, 蒋鹏, 等. 复方扶芳藤合剂含药血清促进骨髓间充质干细胞增殖的作用机制[J]. 器官移植, 2022, 13(3): 363-370. doi: 10.3969/j.issn.1674-7445.2022.03.013
Wu Lingling, Chen Jibing, Jiang Peng, et al. Mechanism of compound Fufangteng mixture-containing serum in promoting proliferation of bone marrow mesenchymal stem cell[J]. ORGAN TRANSPLANTATION, 2022, 13(3): 363-370. doi: 10.3969/j.issn.1674-7445.2022.03.013
Citation: Wu Lingling, Chen Jibing, Jiang Peng, et al. Mechanism of compound Fufangteng mixture-containing serum in promoting proliferation of bone marrow mesenchymal stem cell[J]. ORGAN TRANSPLANTATION, 2022, 13(3): 363-370. doi: 10.3969/j.issn.1674-7445.2022.03.013

复方扶芳藤合剂含药血清促进骨髓间充质干细胞增殖的作用机制

doi: 10.3969/j.issn.1674-7445.2022.03.013
基金项目: 

广西省自然科学基金 2020GXNSFAA238024

广西壮族自治区中青年教师基础能力提升项目 2017KY0311

广西中医药大学研究生教育创新计划项目 YCSW2021232

详细信息
    作者简介:
    通讯作者:

    高宏君,男,1964年生,博士,主任医师,研究方向为器官移植,Email:gao4056@163.com

  • 中图分类号: R617, R285

Mechanism of compound Fufangteng mixture-containing serum in promoting proliferation of bone marrow mesenchymal stem cell

More Information
  • 摘要:   目的  探讨复方扶芳藤合剂含药血清对骨髓间充质干细胞(BMSC)增殖的影响及其机制。  方法  通过直接贴壁法体外分离、培养、纯化大鼠BMSC,对其进行细胞形态学观察,采用流式细胞仪对其表面标志物进行鉴定。复方扶芳藤合剂按3 mL/(kg·d)给予大鼠灌胃14 d后取含药血清。将BMSC分为空白对照组、含药血清组、Notch1小干扰核糖核酸(siRNA)组和Notch1 siRNA+含药血清组,检测各组BMSC的增殖率以及Notch1信号通路相关信使核糖核酸(mRNA)和蛋白的相对表达量。  结果  镜下观察可见第1代BMSC呈长梭形,以平行排列生长为主或呈漩涡状生长。第3代BMSC阳性表达CD90、CD44,而CD45呈阴性表达。与空白对照组比较,含药血清组和Notch1 siRNA+含药血清组BMSC的增殖率均升高,Notch1 siRNA组BMSC的增殖率下降(均为P < 0.05);与Notch1 siRNA组比较,Notch1 siRNA+含药血清组BMSC的增殖率升高(P < 0.05)。与空白对照组比较,含药血清组Hey1、Delta样配体(DLL)1的mRNA和蛋白相对表达量均升高,Notch1 siRNA组和Notch1 siRNA+含药血清组Hey1、DLL1的mRNA和蛋白相对表达量均下降,差异均有统计学意义(均为P < 0.05);与Notch1 siRNA组比较,Notch1 siRNA+含药血清组Hey1、DLL1的mRNA和蛋白相对表达量均升高,差异均有统计学意义(均为P < 0.05)。  结论  复方扶芳藤合剂含药血清可促进大鼠BMSC的增殖,其作用机制可能与Notch1信号通路激活有关。

     

  • 图  1  BMSC的形态学观察

    注:A图为BMSC培养48 h结果;B图为BMSC培养96 h结果;C图为BMSC培养144 h结果;D图为BMSC第1代培养结果(×40)。

    Figure  1.  Morphological observation of BMSC

    图  2  BMSC表面标志物的流式细胞检测结果

    注:A图为BMSC的均一性检测;B图示CD44呈阳性表达,阳性率为86.95 %,C图示CD45呈阴性表达,阳性率为9.90 %;D图示CD90呈阳性表达,阳性率为98. 22 %。

    Figure  2.  Flow cytometry results of surface markers of BMSC

    图  3  复方扶芳藤合剂含药血清对BMSC增殖率的影响

    注:与空白对照组比较,aP < 0.05;与Notch1 siRNA组比较,bP < 0.05。

    Figure  3.  Effect of compound Fufangteng mixture-containing serum on BMSC proliferation rate

    图  4  各组Hey1、DLL1 mRNA相对表达量的比较

    注:与空白对照组比较,aP < 0.05;与Notch1 siRNA组比较,bP < 0.05。

    Figure  4.  Comparison of the relative expression levels of Hey1 and DLL1 mRNA among each group

    图  5  各组Hey1、DLL1蛋白相对表达量的比较

    注:与空白对照组比较,aP < 0.05;与Notch1 siRNA组比较,bP < 0.05。

    Figure  5.  Comparison of the relative expression levels of Hey1 and DLL1 proteins among each group

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  • 收稿日期:  2022-01-19
  • 网络出版日期:  2022-04-29
  • 刊出日期:  2022-05-15

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