支链氨基酸氨基转移酶1通过P53信号调节脂肪间充质干细胞存活

    Branched chain amino acid aminotransferase-1 regulates adipose-derived mesenchymal stem cell survival via P53 signaling.

      摘要
    • 摘要:
        目的  研究支链氨基酸氨基转移酶1(branched-chain-amino-acid aminotransferase 1，BCAT1)调节间充质干细胞存活的作用并探讨机制。
        方法  从雄性Sprague Dawley大鼠附睾白色脂肪中分离脂肪间充质干细胞（adipose-derived mesenchymal stem cells，ADSCs）。腺病毒转染ADSCs中分别过表达或敲低BCAT1表达后给予过氧化氢（hydrogen dioxide，H2O2）刺激诱导ADSCs凋亡。利用CCK-8细胞活力分析、cleaved caspase-3表达及TUNEL染色评估ADSCs凋亡。利用P53特异性抑制剂PFT-1α探讨P53信号在BCAT1调节ADSCs生存的分子机制。
        结果  BCAT1而不是BCAT2是ADSCs的主要表达亚型。H2O2刺激诱导ADSCs凋亡时BCAT1蛋白表达水平显著降低（P<0.01）。使用shRNA敲低BCAT1表达加重而过表达BCAT1显著减轻H2O2诱导的ADSCs凋亡（均P<0.01）。敲低BCAT1导致H2O2刺激的ADSCs中P53下游靶基因表达水平显著增高，提示P53信号活化（均P<0.01）。PFT-1α可以逆转BCAT1敲低恶化的ADSCs凋亡（均P<0.01）。
        结论  BCAT1通过P53信号调节ADSCs生存。BCAT1可作为促进ADSCs存活、增强其心肌保护作用的候选靶点。

       

      Abstract:
        AIM  To determine the regulatory role of branched chain amino acid aminotransferase-1 (BCAT1) in the survival of adipose-derived mesenchymal stem cells (ADSCs).
        METHODS  ADSCs were isolated from white adipose tissues of male Sprague Dawley rats. BCAT1 expression was knocked down or over-expressed in ADSCs by adenovirus transfection and ADSC apoptosis was induced by hydrogen dioxide stimulation. ADSC apoptosis was evaluated by CCK-8 cell viability assay, cleaved caspase-3 expression, and TUNEL staining. The activity of P53 was suppressed by the specific inhibitor PFT-1α in control or BCAT1 silencing ADSCs.
        RESULTS  BCAT1, rather than BCAT2, was the predominant subtype expressed in ADSCs. Silencing of BCAT1 exacerbated, whereas BCAT1 over-expression ameliorated hydrogen dioxide-induced ADSC apoptosis. The downstream targets of P53 were significantly increased in BCAT1 knock-downed ADSCs when stimulated by hydrogen dioxide. Inhibition of P53 by its specific inhibitor PFT-1α reversed the adverse impact of BCAT1 silencing on ADSC survival.
        CONCLUSION  BCAT1 regulates ADSC survival via a P53-dependent manner, revealing that BCAT1 is a promising target to enhance ADSC survival and their cardioprotective efficacy.

       

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