实用医学杂志 ›› 2022, Vol. 38 ›› Issue (6): 726-737.doi: 10.3969/j.issn.1006⁃5725.2022.06.015

• 基础研究 • 上一篇    下一篇

川芎嗪对脐带间充质干细胞移植缺血性脑卒中迁移率的影响及作用机制

曹慧玲 汪小蓉 朱小飞 张洁 钱世宁   

  1. 南京中医药大学附属医院检验科(南京 210029)

  • 出版日期:2022-03-25 发布日期:2022-03-25
  • 基金资助:
    国家自然科学基金青年项目(编号:81703830)

Effect and mechanism of Tetramethylpyrazine on the migration of umbilical cord mesenchymal stem cell in the treatment of ischemic stroke

CAO Huiling,WANG Xiaorong,ZHU Xiaofei,ZHANG Jie,QIAN Shining.   

  1. Department of Clinical Laboratory,the Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China

  • Online:2022-03-25 Published:2022-03-25

摘要:

目的 观察体内外川芎嗪(TMP)对脐带间充质干细胞(ucMSCs)迁移的影响并研究其相关作用机制。方法 体内尾静脉移植ucMSCs 入脑中动脉栓塞模型(MCAO)大鼠,荧光显微镜观察GFP⁃ucMSCs 的迁移定位情况,Western blot检测基质细胞衍生因子⁃1(SDF⁃1)及其受体CXCR4的表达;体外采用Transwell 法检测 ucMSCs 的迁移,观察 TMP 处理以及加入拮抗剂后对 ucMSCs 迁移率的影响,ELISA 检测白介素⁃1β (IL⁃1β)的表达水平,检测迁移相关蛋白表达。结果 体内结果发现 TMP 预处理后 ucMSCs 迁移至脑梗死 区的数量(IOD 2.9 × 105)较未处理组(IOD 0.4 × 105)显著增加(P < 0.05),SDF⁃1 CXCR4 的表达量也显著 升高(P < 0.05),且加入拮抗剂后各指标均明显降低(P < 0.05);体外实验显示 TMP 预处理后的 ucMSCs 移细胞数由每区域 20 个提高至 88 个(P < 0.05),与 CXCR4 的表达升高一致;TMP 处理损伤 PC12 细胞可有 效降低炎症因子 IL⁃1β 的表达(17.2 pg/mL 降至 13.5 pg/mL)。结论 川芎嗪通过调节 SDF⁃1/CXCR4 轴提高 脐带间充质干细胞移植迁移率,并抑制炎症反应改善移植微环境,为中医药与干细胞联合治疗缺血性脑 卒中提供实验依据。

关键词:

川芎嗪, 脐带间充质干细胞, 脑卒中, 移植, 迁移

Abstract:

Objective To observe the effect of tetramethylpyrazine(TMP)on the migration of umbilical cord mesenchymal stem cells(ucMSCs)and investigate the mechanism in vivo and vitro separately. Methods In vivo,ucMSCs were transplanted into MCAO rats by tail vein injection. The migration and localization of GFP⁃ ucMSCs were observed by fluorescence microscope. The expression of related proteins SDF ⁃ 1 and CXCR4 were detected by Western blot. In vitro,the migration of ucMSCs was detected by Transwell method.The effects of TMP treatment and antagonist on the mobility of ucMSCs were observed. The level of interleukin⁃1β(IL⁃1β)was detected by ELISA and the expression of migration related proteins was detected. Results In vivo,it was found that the number of ucMSCs migrated and located in the cerebral infarction area after TMP pretreatment(IOD 2.9 × 105 increased significantly compared to untreated group(IOD 0.4 × 105)(P < 0.05). The expression of SDF⁃1 and CXCR4 also increased significantly,which was inhibited by adding antagonists(P < 0.05);In vitro,the number of migrating cells increased from 20 to 88 per region by TMP pretreatment(P < 0.05),which was consistent with the increased expression of CXCR4. The expression of inflammatory factor IL⁃1 β in PC12 cells was reduced by TMP treatment(from 17.2 to 13.5 pg/mL). Conclusion Tetramethylpyrazine improves the migration rate of umbilical cord mesenchymal stem cells by regulating SDF ⁃ 1/CXCR4 axis.The transplantation microenvironment was also improved by inhibiting inflammation. This study provides experimental basis for the combination of traditional Chinese medicine and stem cells in the treatment of ischemic stroke.

Key words: tetramethylpyrazine,  , umbilical cord mesenchymal stem cells,  , cerebral ischemia?reperfu? sion,  , transplantation,  , migration