留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

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

单佳柔 尼贝贝 李翠平 徐睿璇 陈文捷

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
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

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

doi: 10.3969/j.issn.1674-7445.2021.03.007
基金项目: 

国家自然科学基金 81770651

中山大学青年教师培育项目 19ykpy28

详细信息
    作者简介:

    单佳柔,女,1995年生,硕士研究生,研究方向为生物治疗,Email:shanjiarou@hotmail.com

    通讯作者:

    陈文捷,女,1978年生,博士,副研究员,研究方向为生物治疗,Email: chenwenjie168@aliyun.com

  • 中图分类号: R617

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

More Information
  • 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。

     

  • 图  1  各组L02细胞凋亡率和细胞内ROS相对量的比较

    注:A图为空白对照组和L02+HUC-MSC组的流式细胞图;B图为各组L02细胞凋亡率的比较;C图为各组L02细胞内ROS相对量的比较;与空白对照组比较,aP < 0.05,与OGD组比较,bP < 0.05。

    Figure  1.  Comparison of cell apoptosis rate and intracellular relative ROS level of L02 cells among each group

    图  2  HUC-MSC向L02转移线粒体的流式细胞学表现

    注:A图为流式细胞术检测HUC-MSC与L02;B图为各组HUC-MSC向L02转移线粒体的情况,圈门表示L02细胞群;C图为通过L02细胞群中MitoTracker阳性L02的比例来评估线粒体转移率,与实验对照组比较,aP < 0.05。

    Figure  2.  Flow cytometry findings of mitochondrial transfer from HUC-MSC to L02 cells

    图  3  HUC-MSC向L02转移线粒体的激光共聚焦显微镜下表现

    注:A图示MitoTracker Deep Red标记的HUC-MSC(×200);B图示CellTrace Violet标记的L02(×200);C图示Phaliodin-ifluor 488染色的F-actin(×200);D图(×200)、E图(×400)示HUC-MSC和L02共培养后TNT形成和线粒体转移。

    Figure  3.  Mitochondrial transfer from HUC-MSC to L02 cells under the confocal laser scanning microscopy

  • [1] HEYLEN L, PIRENNE J, NAESENS M, et al. "Time is tissue"-a minireview on the importance of donor nephrectomy, donor hepatectomy, and implantation times in kidney and liver transplantation[J]. Am J Transplant, 2021, DOI: 10.1111/ajt.16580[Epub ahead of print].
    [2] 谷带利, 谢智钦, 唐才喜. 人抗原R: 小鼠和人类肝移植损伤中一种新的血红素氧合酶-1细胞保护调节剂[J]. 临床肝胆病杂志, 2020, 36(4): 931. doi: 10.3969/j.issn.1001-5256.2020.04.060

    GU DL, XIE ZX, TANG CX. Human antigen R(HuR): a new regulator of heme oxygenase-2 cytoprotection in mouse and human liver transplant injury[J]. J Clin Hepatol, 2020, 36(4): 931. doi: 10.3969/j.issn.1001-5256.2020.04.060
    [3] LIN MJ, LI S, YANG LJ, et al. Plasma membrane vesicles of human umbilical cord mesenchymal stem cells ameliorate acetaminophen-induced damage in HepG2 cells: a novel stem cell therapy[J]. Stem Cell Res Ther, 2020, 11(1): 225. DOI: 10.1186/s13287-020-01738-z.
    [4] HU C, WU Z, LI L. Mesenchymal stromal cells promote liver regeneration through regulation of immune cells[J]. Int J Biol Sci, 2020, 16(5): 893-903. DOI: 10.7150/ijbs.39725.
    [5] YOU Y, WEN DG, GONG JP, et al. Research status of mesenchymal stem cells in liver transplantation[J]. Cell Transplant, 2019, 28(12): 1490-1506. DOI: 10.1177/0963689719874786.
    [6] HU C, LI L. The immunoregulation of mesenchymal stem cells plays a critical role in improving the prognosis of liver transplantation[J]. J Transl Med, 2019, 17(1): 412. DOI: 10.1186/s12967-019-02167-0.
    [7] SOUNDARA RAJAN T, GUGLIANDOLO A, BRAMANTI P, et al. Tunneling nanotubes-mediated protection of mesenchymal stem cells: an update from preclinical studies[J]. Int J Mol Sci, 2020, 21(10): 3481. DOI: 10.3390/ijms21103481.
    [8] FENG Y, ZHU R, SHEN J, et al. Human bone marrow mesenchymal stem cells rescue endothelial cells experiencing chemotherapy stress by mitochondrial transfer via tunneling nanotubes[J]. Stem Cells Dev, 2019, 28(10): 674-682. DOI: 10.1089/scd.2018.0248.
    [9] HAN D, ZHENG X, WANG X, et al. Mesenchymal stem/stromal cell-mediated mitochondrial transfer and the therapeutic potential in treatment of neurological diseases[J]. Stem Cells Int, 2020: 8838046. DOI: 10.1155/2020/8838046.
    [10] MOHAMMADALIPOUR A, DUMBALI SP, WENZEL PL. Mitochondrial transfer and regulators of mesenchymal stromal cell function and therapeutic efficacy[J]. Front Cell Dev Biol, 2020, 8: 603292. DOI: 10.3389/fcell.2020.603292.
    [11] RUSSO L, GRACIA-SANCHO J, GARCÍA-CALDERÓ H, et al. Addition of simvastatin to cold storage solution prevents endothelial dysfunction in explanted rat livers[J]. Hepatology, 2012, 55(3): 921-930. DOI: 10.1002/hep.24755.
    [12] SELZNER N, LIU H, BOEHNERT MU, et al. FGL2/fibroleukin mediates hepatic reperfusion injury by induction of sinusoidal endothelial cell and hepatocyte apoptosis in mice[J]. J Hepatol, 2012, 56(1): 153-159. DOI: 10.1016/j.jhep.2011.05.033.
    [13] ZHANG Z, JIN X, YANG C, et al. Teneligliptin protects against hypoxia/reoxygenation-induced endothelial cell injury[J]. Biomed Pharmacother, 2019, 109: 468-474. DOI: 10.1016/j.biopha.2018.10.016.
    [14] HUANG H, TOHME S, AL-KHAFAJI AB, et al. Damage-associated molecular pattern-activated neutrophil extracellular trap exacerbates sterile inflammatory liver injury[J]. Hepatology, 2015, 62(2): 600-614. DOI: 10.1002/hep.27841.
    [15] BAKIRI L, HAMACHER R, GRAÑA O, et al. Liver carcinogenesis by FOS-dependent inflammation and cholesterol dysregulation[J]. J Exp Med, 2017, 214(5): 1387-1409. DOI: 10.1084/jem.20160935.
    [16] YAO J, ZHENG J, CAI J, et al. Extracellular vesicles derived from human umbilical cord mesenchymal stem cells alleviate rat hepatic ischemia-reperfusion injury by suppressing oxidative stress and neutrophil inflammatory response[J]. FASEB J, 2019, 33(2): 1695-1710. DOI: 10.1096/fj.201800131RR.
    [17] SUN Y, WANG Y, ZHOU L, et al. Spheroid-cultured human umbilical cord-derived mesenchymal stem cells attenuate hepatic ischemia-reperfusion injury in rats[J]. Sci Rep, 2018, 8(1): 2518. DOI: 10.1038/s41598-018-20975-0.
    [18] POURMOHAMMADI-BEJARPASI Z, ROUSHANDEH AM, SABERI A, et al. Mesenchymal stem cells-derived mitochondria transplantation mitigates I/R-induced injury, abolishes I/R-induced apoptosis, and restores motor function in acute ischemia stroke rat model[J]. Brain Res Bull, 2020, 165: 70-80. DOI: 10.1016/j.brainresbull.2020.09.018.
    [19] BI ZM, ZHOU QF, GENG Y, et al. Human umbilical cord mesenchymal stem cells ameliorate experimental cirrhosis through activation of keratinocyte growth factor by suppressing microRNA-199[J]. Eur Rev Med Pharmacol Sci, 2016, 20(23): 4905-4912.
    [20] AMIN MA, SABRY D, RASHED LA, et al. Short-term evaluation of autologous transplantation of bone marrow-derived mesenchymal stem cells in patients with cirrhosis: Egyptian study[J]. Clin Transplant, 2013, 27(4): 607-612. DOI: 10.1111/ctr.12179.
    [21] GHAVAMZADEH A, SOTOUDEH M, HASHEMI TAHERI AP, et al. Liver fibrosis alleviation after co-transplantation of hematopoietic stem cells with mesenchymal stem cells in patients with thalassemia major[J]. Ann Hematol, 2018, 97(2): 327-334. DOI: 10.1007/s00277-017-3181-9.
    [22] SHIN B, COWAN DB, EMANI SM, et al. Mitochondrial transplantation in myocardial ischemia and reperfusion injury[J]. Adv Exp Med Biol, 2017, 982: 595-619. DOI: 10.1007/978-3-319-55330-6_31.
    [23] GO KL, LEE S, ZENDEJAS I, et al. Mitochondrial dysfunction and autophagy in hepatic ischemia/reperfusion injury[J]. Biomed Res Int, 2015: 183469. DOI: 10.1155/2015/183469.
    [24] MUI D, ZHANG Y. Mitochondrial scenario: roles of mitochondrial dynamics in acute myocardial ischemia/reperfusion injury[J]. J Recept Signal Transduct Res, 2021, 41(1): 1-5. DOI: 10.1080/10799893.2020.1784938.
    [25] GU J, ZHANG T, GUO J, et al. PINK1 Activation and translocation to mitochondria-associated membranes mediates mitophagy and protects against hepatic ischemia/reperfusion injury[J]. Shock, 2020, 54(6): 783-793. DOI: 10.1097/SHK.0000000000001534.
    [26] DENG F, WANG S, ZHANG L, et al. Propofol through upregulating caveolin-3 attenuates post-hypoxic mitochondrial damage and cell death in H9C2 cardiomyocytes during hyperglycemia[J]. Cell Physiol Biochem, 2017, 44(1): 279-292. DOI: 10.1159/000484680.
    [27] PARADIES G, PARADIES V, RUGGIERO FM, et al. Mitochondrial bioenergetics and cardiolipin alterations in myocardial ischemia-reperfusion injury: implications for pharmacological cardioprotection[J]. Am J Physiol Heart Circ Physiol, 2018, 315(5): H1341-H1352. DOI: 10.1152/ajpheart.00028.2018.
    [28] LESNEFSKY EJ, CHEN Q, TANDLER B, et al. Mitochondrial dysfunction and myocardial ischemia-reperfusion: implications for novel therapies[J]. Annu Rev Pharmacol Toxicol, 2017, 57: 535-565. DOI: 10.1146/annurev-pharmtox-010715-103335.
    [29] JOHNSON K, MCEVOY CE, NAQVI S, et al. High-dose oral N-acetylcysteine fails to improve respiratory health status in patients with chronic obstructive pulmonary disease and chronic bronchitis: a randomized, placebo-controlled trial[J]. Int J Chron Obstruct Pulmon Dis, 2016, 11: 799-807. DOI: 10.2147/COPD.S102375.
    [30] LI X, MICHAELOUDES C, ZHANG Y, et al. Mesenchymal stem cells alleviate oxidative stress-induced mitochondrial dysfunction in the airways[J]. J Allergy Clin Immunol, 2018, 141(5): 1634-1645. DOI: 10.1016/j.jaci.2017.08.017.
  • 加载中
图(5)
计量
  • 文章访问数:  175
  • HTML全文浏览量:  72
  • PDF下载量:  29
  • 被引次数: 0
出版历程
  • 收稿日期:  2020-12-17
  • 网络出版日期:  2021-05-19
  • 刊出日期:  2021-05-15

目录

    /

    返回文章
    返回