Tang Mao-Han, Zhang Yan-Jie, Liu Yin, Su Zhi-Guang
Molecular Medicine Research Center, West China Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2017 Mar;48(2):173-178.
To investigate the effects of trimetazidine (TMZ) on the oxidative stress injury in adipose-derived mesenchymal stem cells (ADSCs).
ADSCs derived from adipose tissue of SD rats were characterized by flow cytometry and multiline age differentiation. ADSCs apoptosis was induced by H2O2 , Dirrerent concentration of TMZ (250 μmol/L, 500 μmol/L) was used to protect ADSCs from apoptosis. The morphological features of apoptotic ADSCs were analyzed by Hoechst 33342, mitochondrial potential and structure was analyzed by JC-1 staining and electron microscope, respectively. The apoptotic proteins were detected by Western blot. The effect of TMZ on antioxidant capacity of ADSCs was evaluated by detecting reactive oxygen species (ROS), superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA).
The isolated ADSCs expressed high levels of CD29 and CD90, low levels of CD34 and CD45 and no expression of CD31. ADSCs could be induced to adipocyte and osteoplastic cells. After being treated by H2O2, ADSCs displayed apoptosis characteristics with increased number of apoptotic cells, decreased mitochondrial transmembrane potential and damaged mitochondria. The expressions of apoptotic proteins, including Bax, Bad, and Caspase3, were dramatically increased compared to the controls; however, the anti-apoptotic protein Bcl2 was decreased. At the meantime, the contents of ROS and MDA were elevated, but the concentrations of SOD and GSH were reduced. The treatment of TMZ could partly reverse above negative impacts to ADSCs.
TMZ could improve the survival rate of ADSCs by enhancing anti-oxidant defense systems to remove excessive ROS and regulating the expression of protective protein.
研究曲美他嗪（TMZ）对脂肪来源间充质干细胞（ADSCs）氧化应激损伤的影响。
采用流式细胞术和多向分化对SD大鼠脂肪组织来源的ADSCs进行鉴定。用H2O2诱导ADSCs凋亡，使用不同浓度的TMZ（250μmol/L、500μmol/L）保护ADSCs免于凋亡。分别用Hoechst 33342分析凋亡ADSCs的形态特征，用JC-1染色和电子显微镜分析线粒体电位和结构。通过蛋白质免疫印迹法检测凋亡蛋白。通过检测活性氧（ROS）、超氧化物歧化酶（SOD）、谷胱甘肽（GSH）和丙二醛（MDA）评估TMZ对ADSCs抗氧化能力的影响。
分离的ADSCs高表达CD29和CD90，低表达CD34和CD45，不表达CD31。ADSCs可被诱导分化为脂肪细胞和成骨细胞。H2O2处理后，ADSCs呈现凋亡特征，凋亡细胞数量增加，线粒体跨膜电位降低，线粒体受损。与对照组相比，凋亡蛋白Bax、Bad和Caspase3的表达显著增加；然而，抗凋亡蛋白Bcl2减少。同时，ROS和MDA含量升高，但SOD和GSH浓度降低。TMZ处理可部分逆转上述对ADSCs的负面影响。
TMZ可通过增强抗氧化防御系统清除过量ROS和调节保护性蛋白表达来提高ADSCs的存活率。
