Chen Ruolin, Liu Shuang, Piao Fengyuan, Wang Zhemin, Qi Yuan, Li Shuangyue, Zhang Dongmei, Shen Jingshun
Department of Occupational and Environmental Health, Dalian Medical University, China.
Ind Health. 2015;53(3):222-35. doi: 10.2486/indhealth.2014-0182. Epub 2015 Jan 29.
2,5-hexanedione (HD) induces apoptosis of nerve cells. However,the mechanism of HD-induced apoptosis remains unknown. Mesenchymal stem cells (MSCs) are multipotential stem cells with the ability to differentiate into various cell types. This study is designed to investigate the apoptosis induced by HD in rat bone marrow MSCs (BMSCs) and the related underlying mechanisms. The fifth generation of MSCs was treated with 0, 10, 20 and 40 mM HD respectively. The viability of BMSCs was observed by MTT. Apoptosis were estimated by Hoechst 33342 staining and TUNEL assay. The disruption of mitochondrial transmembrane potential (MMP) was examined by JC-1 staining. Moreover, the expression of Bax and Bcl-2, cytochrome c release, and caspase-3 activity were determined by real-time RT-PCR, Western blot and Spectrophotometry. Our results showed that HD induced apoptosis in MSCs in a dose dependent manner. Moreover, HD downregulated the Bcl-2 expression,upregulated the Bax expression and the Bax/Bcl-2 ratio, promoted the disruption of MMP, induced the release of cytochrome c from mitochondria to cytosol, and increased the activity of caspase-3 in MSCs. These results indicate that HD induces apoptosis in MSCs and the activated mitochondria-dependent caspase-3 pathway may be involved in the HD-induced apoptosis.
2,5-己二酮（HD）可诱导神经细胞凋亡。然而，HD诱导凋亡的机制尚不清楚。间充质干细胞（MSCs）是具有分化为多种细胞类型能力的多能干细胞。本研究旨在探讨HD诱导大鼠骨髓间充质干细胞（BMSCs）凋亡及其相关潜在机制。分别用0、10、20和40 mM HD处理第五代MSCs。通过MTT观察BMSCs的活力。通过Hoechst 33342染色和TUNEL检测评估凋亡情况。通过JC-1染色检测线粒体跨膜电位（MMP）的破坏。此外，通过实时RT-PCR、Western印迹和分光光度法测定Bax和Bcl-2的表达、细胞色素c的释放以及caspase-3的活性。我们的结果表明，HD以剂量依赖性方式诱导MSCs凋亡。此外，HD下调Bcl-2表达，上调Bax表达和Bax/Bcl-2比值，促进MMP破坏，诱导细胞色素c从线粒体释放到细胞质中，并增加MSCs中caspase-3的活性。这些结果表明，HD诱导MSCs凋亡，激活的线粒体依赖性caspase-3途径可能参与HD诱导的凋亡。
