中国血液净化

中国血液净化 ›› 2016, Vol. 15 ›› Issue (04): 226-229.doi: 10.3969/j.issn.1671-4091.2016.04.009

• 基础研究 • 上一篇    下一篇

腹膜透析液对腹膜间皮细胞早衰的影响及间充质干细胞干预的研究

曹东维,张苗,蒋春明,孙琤   

  1. 南京大学医学院附属鼓楼医院肾脏内科
  • 收稿日期:2015-12-17 修回日期:2016-01-24 出版日期:2016-04-12 发布日期:2016-04-19
  • 通讯作者: 张苗 zmslp@medmail.com.cn E-mail:zmslp@medmail.com.cn
  • 基金资助:

    中央高校基本科研业务费专项资金项目(021414340282)

Studies on the senescent processes of human peritoneal mesothelial cells relating to peritoneal dialysis fluid and the intervention effect of bone marrow derived mesenchymal stem cells

  • Received:2015-12-17 Revised:2016-01-24 Online:2016-04-12 Published:2016-04-19

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摘要： 目的探讨腹膜透析液对腹膜间皮细胞早衰的影响及骨髓间充质干细胞的干预作用。方法首先从择期行腹膜透析置管手术者大网膜分离培养出腹膜间皮细胞。将腹膜透析液加入常规培养的腹膜间皮细胞，通过腹膜间皮细胞传代数、细胞增殖、细胞周期、端粒的检测以观察腹膜透析液对腹膜间皮细胞衰老的影响及可能机制；通过在培养液中加入骨髓间充质干细胞，观察骨髓间充质干细胞对腹膜间皮细胞早衰的干预作用。结果与仅用RPMI1640 培养基培养的腹膜间皮细胞相比，腹膜透析液致腹膜间皮细胞传代数减少4～5 代，生长速率降低42%，细胞周期阻滞于G1 期[(69.1 ± 0.3)%]，端粒长度缩短(4.35 ± 0.12) kb 比(5.45 ± 0.10) kb, t= 2.228，P=0.000]。腹膜透析患者透出液中分离培养出腹膜的间皮细胞接近衰老细胞，加入骨髓间充质干细胞培养的间皮细胞衰老进程延缓。结论骨髓间充质干细胞可延缓腹膜透析液所诱导的腹膜间皮细胞的衰老进程。为临床上延缓腹膜纤维化，改善腹膜透析的远期疗效提供一种新的理论依据和治疗手段。

关键词: 腹膜透析液, 细胞衰老, 腹膜间皮细胞, 间充质干细胞

Abstract: Objective To investigate the replicative senescence of human peritoneal mesothelial cells (HPMCs) induced by peritoneal dialysis fluid (PDF) and the intervention effect of bone marrow derived mesenchymalstem cells (MSCs). Methods HPMCs were isolated from peritoneal tissue and cultured in RPMI
1640 medium. PDF was added to the cultured HPMCs. The effects of PDF on senescence of HPMCs were examined by observing number of cell passages, cell proliferation, cell cycle and telomere length. After the addition of MSCs into the culture medium, the effects of MSCs on the senescence of HPMCs were observed, and the related mechanism was evaluated by observing number of cell passages, cell growth rate, cell cycle and telomere length. Results PDF treated HPMCs showed reduction of cell passage number by four to five, suppression of growth rate by 42%, and cease of cell cycle at G1 phase, as compared with the HPMCs cultured in RPMI 1640. Telomere length was shortened (4.35±0.12 kb vs. 5.45±0.10 kb, t=2.228, P=0.000]. Morphological characteristics of senescent cells appeared earlier. The senescence process delayed in HPMCs with the addition of MSCs. Conclusion MSCs may delay cellular senescence of the HPMCs induced by PDF treatment, which provides a new theoretical basis and a treatment method for delaying peritoneal fibrosis and improving longterm efficacy of peritoneal dialysis.

Key words: Peritoneal dialysis fluid, cellular senescence, HPMCs, MSCs