单佳柔, 尼贝贝, 李翠平, 等. 人脐带间充质干细胞通过线粒体转移减轻肝细胞缺血-再灌注损伤的机制研究[J]. 器官移植, 2021, 12(3): 294-301. DOI: 10.3969/j.issn.1674-7445.2021.03.007
引用本文: 单佳柔, 尼贝贝, 李翠平, 等. 人脐带间充质干细胞通过线粒体转移减轻肝细胞缺血-再灌注损伤的机制研究[J]. 器官移植, 2021, 12(3): 294-301. DOI: 10.3969/j.issn.1674-7445.2021.03.007
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Shan Jiarou, Ni Beibei, Li Cuiping, et al. Mechanism of human umbilical cord mesenchymal stem cells alleviating ischemia-reperfusion injury of hepatocytes through mitochondrial transfer[J]. ORGAN TRANSPLANTATION, 2021, 12(3): 294-301. DOI: 10.3969/j.issn.1674-7445.2021.03.007
Citation: Shan Jiarou, Ni Beibei, Li Cuiping, et al. Mechanism of human umbilical cord mesenchymal stem cells alleviating ischemia-reperfusion injury of hepatocytes through mitochondrial transfer[J]. ORGAN TRANSPLANTATION, 2021, 12(3): 294-301. DOI: 10.3969/j.issn.1674-7445.2021.03.007
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人脐带间充质干细胞通过线粒体转移减轻肝细胞缺血-再灌注损伤的机制研究

Mechanism of human umbilical cord mesenchymal stem cells alleviating ischemia-reperfusion injury of hepatocytes through mitochondrial transfer

    摘要
  • 摘要:
      目的  探讨人脐带间充质干细胞（HUC-MSC）通过线粒体转移减轻肝细胞缺血-再灌注损伤（IRI）的作用机制。
      方法  将正常人肝细胞株L02分为空白对照组、氧糖剥夺（OGD）组、实验对照组、L02与HUC-MSC共培养组（L02+HUC-MSC组），其中L02+HUC-MSC组根据L02与HUC-MSC不同共培养比例分为10:1共培养组（A组）、4:1共培养组（B组）、2:1共培养组（C组）、1:1共培养组（D组）和1:2共培养组（E组）。采用流式细胞术检测L02细胞凋亡率和细胞内活性氧簇（ROS）相对量；采用流式细胞术检测L02的MitoTracker阳性率；采用激光共聚焦显微镜观察HUC-MSC向L02转移线粒体的情况。
      结果  OGD组L02细胞凋亡率和细胞内ROS相对量高于空白对照组（均为P < 0.05）；与OGD组比较，B、C、D、E组L02细胞凋亡率均降低（均为P < 0.05），E组L02细胞内ROS相对量降低（P < 0.05）。A组L02的MitoTracker阳性率与实验对照组比较，差异无统计学意义（P>0.05），B、C、D、E组均较实验对照组增加（均为P < 0.05），并呈浓度依赖性。在激光共聚焦显微镜下可观察到线粒体通过隧道纳米管（TNT）从HUC-MSC向L02转移。
      结论  HUC-MSC可通过细胞间直接转移线粒体的方式减轻肝细胞IRI后细胞凋亡和降低细胞内ROS水平。

     

    Abstract:
      Objective  To explore the mechanism of human umbilical cord mesenchymal stem cell (HUC-MSC) alleviating ischemia-reperfusion injury (IRI) of liver cells through mitochondrial transfer.
      Methods  Normal human liver cell line L02 was divided into the blank control group, oxygen-glucose deprivation (OGD) group, experimental control group, and L02 and HUC-MSC co-culture group (L02+HUC-MSC group). L02+HUC-MSC group was further divided into 10:1 co-culture subgroup (group A), 4:1 co-culture subgroup (group B), 2:1 co-culture subgroup (group C), 1:1co-culture subgroup (group D) and 1:2 co-culture subgroup (group E) according to different co-culture ratio of L02 and HUC-MSC. The apoptosis rate and relative reactive oxygen species (ROS) level of L02 cells were detected by flow cytometry. The MitoTracker positive rate of L02 cells was detected by flow cytometry. The mitochondrial transfer from HUC-MSC to L02 cells was observed by laser confocal microscope.
      Results  The apoptosis rate and relative ROS level of L02 cells in the OGD group were significantly higher than those in the blank control group (both P < 0.05). Compared with the OGD group, the apoptosis rates of L02 cells in group B, C, D and E were significantly decreased (all P < 0.05), and the relative ROS level of L02 cells in group E was significantly declined (P < 0.05). The MitoTracker positive rate of L02 cells did not significantly differ between group A and experimental control group (P>0.05), whereas the MitoTracker positive rates of L02 cells in group B, C, D and E were significantly higher than that in the experimental control group in a concentration-dependent manner (all P < 0.05). Under the laser confocal microscope, mitochondrial transfer fromHUC-MSC to L02 cells could be observed through tunneling nanotube (TNT).
      Conclusions  HUC-MSC may alleviate cell apoptosis and reduce ROS level of liver cells after IRI via direct mitochondrial transfer between cells.

     

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