Kharchuk I V, Andrukhova O O, Rybal'chenko V K, Andrukhov O
Fiziol Zh (1994). 2013;59(3):50-7.
The features of the impact of the maleimide derivative 1-(4-Cl-benzyl)-3-chloro-4-(CF3-fenilamino)-1H-pyrrole-2,5-dione (MI-1) on the viability and apoptosis-induced cell death of renal proximal and distal tubular epithelial cells and the amount of total and phosphorylated ERK1/2 were studied in order to establish possible mechanisms of nephrotoxicity induced by of MI-1. The viability and apoptosis of renal epithelial tubular cells after incubation with MI-1 were perfomed by 3,4,5-dymetyltiazol-2-yl-2,5-diphenyl-tetrazolium bromide (MTT)-test and by flow cytometry after staining with specific antibodies to annexin V, respectively. The amount of ERK 1/2 was determined by Western blotting. The data indicate that MI-1 was more toxic with respect to the epithelial cells of distal than proximal tubule cells. The apoptosis-induced cell death pathway is involved in the mechanisms of MI-1 cytotoxicity. One of the possible mechanisms of MI-1 nephrotoxicity is increase in phosphorylation of ERK1/2 in the distal tubules. At the same time the increase amount of total ERK1/2 in proximal tubules under the influence of MI-1 may contribute to the survival of proximal tubular epithelial cells under the impact of a toxic factor or oxidative stress.
为了确定马来酰亚胺衍生物1-(4-氯苄基)-3-氯-4-(三氟苯基氨基)-1H-吡咯-2,5-二酮(MI-1)诱导肾毒性的可能机制，研究了其对肾近端和远端肾小管上皮细胞活力、凋亡诱导的细胞死亡以及总ERK1/2和磷酸化ERK1/2水平的影响。分别通过3,4,5-二甲基噻唑-2-基-2,5-二苯基溴化四氮唑(MTT)试验以及用膜联蛋白V特异性抗体染色后通过流式细胞术检测MI-1孵育后肾上皮肾小管细胞的活力和凋亡情况。通过蛋白质免疫印迹法测定ERK 1/2的水平。数据表明，MI-1对远端肾小管上皮细胞的毒性大于近端肾小管上皮细胞。凋亡诱导的细胞死亡途径参与了MI-1细胞毒性机制。MI-1肾毒性的可能机制之一是远端肾小管中ERK1/2磷酸化增加。同时，在MI-1影响下近端肾小管中总ERK1/2水平的增加可能有助于近端肾小管上皮细胞在毒性因子或氧化应激作用下存活。
