Research Center for Cardiovascular Regenerative Medicine, Fuwai Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, 167 Beilishilu, Beijing 100037, People's Republic of China.
J Cell Biochem. 2010 Nov 1;111(4):967-78. doi: 10.1002/jcb.22785.
Poor survival of mesenchymal stem cells (MSCs) compromised the efficacy of stem cell therapy for myocardial infarction. The increase of exogenous reactive oxygen species (ROS) in infracted heart is one of the important factors that challenged the survival of donor MSCs. In the study we aimed to evaluate the effect of oxidative stress on the cell death of MSCs and investigate its mechanisms in order to help with the identification of new biological compounds to reduce donor cells damage. Apoptosis of MSCs were evaluated with Hoechst 33342 staining and flow cytometry analysis. The mitochondrial membrane potential of MSCs was analyzed with JC-1 staining. Signaling pathways involved in H(2)O(2) induced apoptosis were analyzed with Western blot. H(2)O(2) induced apoptosis of MSCs in a dose- and time-dependent manner. H(2)O(2) induced apoptosis of MSCs via both endoplasmic reticulum (ER) and mitochondrial pathways rather than extrinsic apoptosis pathway. H(2)O(2) caused transient rather than sustained activation of p38 and JNK with no effect on ERK1/2 pathway. P38 was involved in the regulation of early apoptosis of MSCs while JNK was involved in the late apoptosis. P38 directed both ER stress and mitochondria death pathway in the early apoptosis. In conclusion, exogenous ROS was a major factor to induce apoptosis of MSCs. Both ER stress and mitochondria death pathway were involved in the apoptosis of MSCs. H(2)O(2) activated p38 that directed the above two pathways in the regulation of early apoptosis of MSCs while JNK was involved in the late apoptosis of MSCs.
间充质干细胞（MSCs）的存活能力较差，这影响了干细胞疗法治疗心肌梗死的效果。梗死心脏中外源活性氧（ROS）的增加是挑战供体 MSC 存活的重要因素之一。本研究旨在评估氧化应激对 MSC 细胞死亡的影响，并探讨其机制，以帮助寻找新的生物化合物来减少供体细胞损伤。通过 Hoechst 33342 染色和流式细胞术分析评估 MSC 的细胞凋亡。通过 JC-1 染色分析 MSC 的线粒体膜电位。通过 Western blot 分析 H2O2 诱导的凋亡相关信号通路。H2O2 以剂量和时间依赖的方式诱导 MSC 凋亡。H2O2 通过内质网（ER）和线粒体途径而非外在凋亡途径诱导 MSC 凋亡。H2O2 引起 p38 和 JNK 的短暂而非持续激活，对 ERK1/2 途径没有影响。p38 参与 MSC 早期凋亡的调节，而 JNK 参与晚期凋亡。p38 调节早期凋亡中 ER 应激和线粒体死亡途径。总之，外源性 ROS 是诱导 MSC 凋亡的主要因素。ER 应激和线粒体死亡途径均参与 MSC 的凋亡。H2O2 激活 p38，在 MSC 早期凋亡的调节中，p38 定向上述两个途径，而 JNK 则参与 MSC 的晚期凋亡。
