Yang Fan, Chen Hongyang, Liu Yanju, Yin Kun, Wang Yang, Li Xingda, Wang Guohui, Wang Siyue, Tan Xueying, Xu Chaoqian, Lu Yanjie, Cai Benzhi
Department of Pharmacology, State-Province Key Laboratories of Biomedicine- Pharmaceutics of China, Harbin Medical University, Harbin, China.
Cell Physiol Biochem. 2013;32(4):1072-82. doi: 10.1159/000354507. Epub 2013 Nov 4.
BACKGROUND/AIMS: Doxorubicin is a widely used chemotherapeutic agent, but its clinical use is restricted because of a high risk of cardiotoxicity. Bone marrow-derived mesenchymal stem cells (BMSCs) may repair ischaemically damaged myocardium through transdifferentiation and paracrine action. The aim of this study is to investigate if doxorubicin causes the apoptosis of BMSCs and in turn impairs its healing ability.
BMSCs were exposed to doxorubicin, and cell apoptosis was determined by western blot and stainings.
Doxorubicin reduced the survival ratio and caused the apoptosis of BMSCs, with the increase of intracellular ROS level and depolarization of mitochondrial membrane potential. The ROS scavenger NAC abrogated these consequences. Moreover, doxorubicin markedly activated phosphorylated ERK, p38 and JNK proteins in BMSCs. The specific inhibitors for p38 (SB203580) and JNK (SP600125) may abolish doxorubicin-induced apoptosis of BMSCs but the specific ERK inhibitor (PD98059) not, indicating p38 and JNK activation contribute to BMSCs apoptosis. Also, the phosphorylated and total p53 proteins were increased in doxorubicin-treated BMSCs. Proapoptotic cleaved caspases-3 was upregulated and antiapoptotic Bcl-2 protein was reduced in doxorubicin-treated BMSCs. At last, ELISA assay showed that doxorubicin treatment reduced the VEGF and IGF-1 released by BMSCs.
Taken together, doxorubicin caused BMSCs apoptosis associated with p38, JNK and p53 pathways.
背景/目的：阿霉素是一种广泛使用的化疗药物，但其临床应用因心脏毒性风险高而受到限制。骨髓间充质干细胞（BMSC）可能通过转分化和旁分泌作用修复缺血损伤的心肌。本研究的目的是调查阿霉素是否会导致BMSC凋亡，进而损害其修复能力。
将BMSC暴露于阿霉素中，通过蛋白质印迹法和染色法测定细胞凋亡情况。
阿霉素降低了BMSC的存活率并导致其凋亡，同时细胞内活性氧（ROS）水平升高，线粒体膜电位去极化。ROS清除剂NAC消除了这些后果。此外，阿霉素显著激活了BMSC中磷酸化的细胞外信号调节激酶（ERK）、p38和c-Jun氨基末端激酶（JNK）蛋白。p38特异性抑制剂（SB203580）和JNK特异性抑制剂（SP600125）可消除阿霉素诱导的BMSC凋亡，但ERK特异性抑制剂（PD98059）则不能，这表明p38和JNK的激活促成了BMSC凋亡。此外，在阿霉素处理的BMSC中，磷酸化和总p53蛋白增加。促凋亡的裂解型半胱天冬酶-3上调，抗凋亡的Bcl-2蛋白减少。最后，酶联免疫吸附测定（ELISA）显示，阿霉素处理降低了BMSC释放的血管内皮生长因子（VEGF）和胰岛素样生长因子-1（IGF-1）。
综上所述，阿霉素导致BMSC凋亡，且与p38、JNK和p53信号通路有关。
