Factors released from embryonic stem cells inhibit apoptosis of H9c2 cells
Factors released from embryonic stem cells inhibit apoptosis of H9c2 cells
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DOI：
10.1152/ajpheart.00431.2007
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发表时间：
2007-09-01
影响因子：
4.8
通讯作者：
McDonald, Debbie E.
中科院分区：
文献类型：
--
作者：
Singla, Dinender K.;McDonald, Debbie E.
表明心肌梗死后移植的胚胎干细胞（ES）分化为心脏中的主要细胞类型并改善心脏功能。然而，与观察到的功能改善相比，再生的程度相对较低。其改善功能的机制完全未知。在这份报告中，我们提供了证据，使用细胞培养模型系统的新机制，涉及释放的细胞保护，抗凋亡因子（S）从ES细胞和抑制H2 O2诱导的大鼠心肌细胞衍生的细胞系H9 C2的凋亡。从用和不用H2 O2处理的生长中的小鼠ES细胞产生条件培养基（CM）。H9 c2细胞暴露于H2 O2 2 2 h后诱导凋亡，随后用新鲜细胞培养物或ES细胞-CM替换。24小时后，用两种ES细胞-CM处理的H9 c2细胞表现出显著降低的凋亡，如通过末端脱氧核苷酸转移酶dUTP介导的缺口末端标记染色、凋亡ELISA、caspase-3活性和DNA梯状条带所确定的。接下来，使用Luminex技术，我们检查了两种ES细胞-CM中抗凋亡蛋白胱抑素C、骨桥蛋白和簇蛋白以及抗纤维化的金属蛋白酶组织抑制剂-1（TIMP-1）的存在。释放的因子水平比H9 c2细胞-CM中的因子水平高2- 170倍。TIMP-1抗体可明显抑制ES细胞CM的抗凋亡作用，提示TIMP-1是抑制细胞凋亡的重要因子。此外，我们使用CM从TIMP-1过表达的细胞系，并证明H2 O2诱导的H9 c2细胞凋亡显着抑制。这些观察结果表明，从ES细胞释放的因子含有抗凋亡因子，其作用由TIMP-1介导。此外，这些研究结果表明，释放的因子可能是有用的治疗应用于缺血性心脏病以及许多其他疾病。
suggests that transplanted embryonic stem (ES) cells subsequent to myocardial infarction differentiate into the major cell types in the heart and improve cardiac function. However, the extent of regeneration is relatively meager compared with the observed functional improvement. The mechanisms underlying their improved function are completely unknown. In this report, we provide evidence using a cell culture model system for novel mechanisms that involve the release of cytoprotective, antiapoptotic factor(s) from ES cells and inhibit H2O2-induced apoptosis in the rat cardiomyocyte-derived cell line H9c2. Conditioned medium (CM) from growing mouse ES cells treated with and without H2O2 was generated. Apoptosis was induced after exposure to H2O2 in H9c2 cells for 2 h followed by replacement with fresh cell culture or ES cell-CM. After 24 h, H9c2 cells treated with both ES cell-CMs demonstrated significantly decreased apoptosis, as determined by terminal deoxynucleotidyl transferase dUTP-mediated nick-end labeling staining, apoptotic ELISA, caspase-3 activity, and DNA ladder. Next, using Luminex technology, we examined the presence of antiapoptotic proteins cystatin c, osteopontin, and clusterin and anti-fibrotic, tissue inhibitor of metalloproteinase-1 (TIMP-1) in both ES cell-CMs. The levels of released factors were 2- to 170-fold higher than those in H9c2 cell-CM. Antiapoptotic effects of ES cell-CM were significantly inhibited with TIMP-1 antibody, suggesting that TIMP-1 is an important factor to inhibit apoptosis. Furthermore, we used CM from an TIMP-1-overexpressing cell line and demonstrated that H2O2-induced apoptosis in the H9c2 cells was significantly inhibited. These observations demonstrate that factors released from ES cells contain antiapoptotic factors and that the effects are mediated by TIMP-1. Moreover, these findings suggest that released factors might be useful for therapeutic applications in ischemic heart disease as well as for many other diseases.