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目的研究在HEK293细胞中过表达代谢型谷氨酸受体1a(Metabotropic gluatamate receptor 1,mGluR1a),引起激动剂非依赖型细胞死亡的分子机制和硫氧还蛋白-1(Trx1)在此过程中的作用。方法采用噻唑蓝法、DCF法、免疫印迹法以及氧化还原蛋白印迹法分别检测了Trx1对细胞活力,细胞内活性氧(ROS)含量,AKT等信号通路的影响,以及Trx1本身氧化还原状态的改变。结果 HEK293细胞中过表达mGluR1a,发现ROS生成量增多,磷酸化AKT减少,PARP剪切量增多,Bcl-2表达量减少以及细胞活力降低。共转染Trx1后,可逆转上述现象。同时发现Trx1、mGluR1a之间不存在相互作用。结论在HEK293细胞中,Trx1通过清除过表达mGluR1a产生的ROS,调节相关的信号通路,从而调控过表达mG luR1a导致的细胞毒性造成的细胞死亡。本文揭示了Trx1,mGluR1a与ROS之间的一种新调节关系,以及这种关系在调控细胞生长和死亡过程中的重要意义。
Objective To investigate the molecular mechanism of over-expression of metabotropic glutamate receptor 1 (mGluR1a) in HEK293 cells, which induces agonist-independent cell death and thioredoxin-1 (Trx1) Role. Methods The effects of Trx1 on cell viability, intracellular reactive oxygen species (ROS), AKT and other signaling pathways, as well as the redox state of Trx1 itself, were examined by thiazolyl blue method, DCF method, Western blotting and redox blotting, respectively . Results The overexpression of mGluR1a in HEK293 cells showed that ROS production increased, phosphorylated AKT decreased, PARP shear increased, Bcl-2 decreased and cell viability decreased. After co-transfection with Trx1, the above phenomenon can be reversed. Also found no interaction between Trx1 and mGluR1a. Conclusion In HEK293 cells, Trx1 regulates cell death caused by over-expressed mGluR1a cytotoxicity by regulating the signaling pathways regulated by overexpression of mGluR1a. This article reveals a new regulatory relationship between Trx1, mGluR1a and ROS, and the significance of this relationship in the regulation of cell growth and death.