Wang Chun-Yu, Tang Bei-Sha, Zhang Hai-Nan, Tang Jian-Guang, Guo Ji-Feng, Yan Xin-Xiang, Tan Li-Ming
Department of Neurology, The Second Xiangya Hospital, Central South University, Changsha 410011, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2010 Jul;41(4):604-8.
To study the effect of alpha-ketoglutarate carrier protein (2-oxoglutarate carrier protein, OGCP) and the Parkin protein on HEK293 cell function.
The cell apoptosis rate, mitochondrial membrane potential and intracellular reactive oxygen species of HEK293 cells treated with rotenone, OGCP and / or Parkin protein were detected by using flow cytometry methods (FCM).
(1) Over-expression wild-type Parkin protein and/or OGCP can increase mitochondrial membrane potential of HEK293 cells induced by rotenone, reduce intracellular reactive oxygen species and cell apoptosis rate of HEK293 cells induced by rotenone, while over-expression mutant Parkin (R42P and T240R) protein can decrease the mitochondrial membrane potential of HEK293 cells, especially the HEK293 cells induced by rotenone, but increase intracellular reactive oxygen species and promote apoptosis. (2) In addition, we also found that OGCP can inhibit the increasing of mitochondrial membrane potential and reactive oxygen species and decreasing of cell apoptosis caused by mutant Parkin protein (R42P and T240R).
(1) Parkin protein and OGCP may be associated with the maintenance of normal function of mitochondria. (2) Over-expression of mutant parkin (R42P and T240R) protein may inhibit mitochondrial function and promote apoptosis. (3) Over-expression OGCP has protective effect on cell toxicity caused by rotenone and mutant parkin protein.
研究α-酮戊二酸载体蛋白（2-氧代戊二酸载体蛋白，OGCP）和帕金蛋白对人胚肾293（HEK293）细胞功能的影响。
采用流式细胞术（FCM）检测用鱼藤酮、OGCP和/或帕金蛋白处理的HEK293细胞的细胞凋亡率、线粒体膜电位和细胞内活性氧。
（1）过表达野生型帕金蛋白和/或OGCP可增加鱼藤酮诱导的HEK293细胞的线粒体膜电位，降低鱼藤酮诱导的HEK293细胞的细胞内活性氧水平和细胞凋亡率，而过表达突变型帕金蛋白（R42P和T240R）可降低HEK293细胞的线粒体膜电位，尤其是鱼藤酮诱导的HEK293细胞，但增加细胞内活性氧水平并促进细胞凋亡。（2）此外，我们还发现OGCP可抑制突变型帕金蛋白（R42P和T240R）引起的线粒体膜电位升高、活性氧增加和细胞凋亡减少。
（1）帕金蛋白和OGCP可能与线粒体正常功能的维持有关。（2）突变型帕金蛋白（R42P和T240R）的过表达可能抑制线粒体功能并促进细胞凋亡。（3）OGCP的过表达对鱼藤酮和突变型帕金蛋白引起的细胞毒性具有保护作用。
