She X R, Tian X, Fan X K, Hong G L, Zhao G J, Li M F, Lu Z Q
The First College of Clinical Medical Science, China Three Gorges University, Yichang 443000, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2016 Nov 20;34(11):805-809. doi: 10.3760/cma.j.issn.1001-9391.2016.11.002.
To investigate the protective effect of P-glycoprotein up-regulated by ulinastatin (UTI) on HK-2 cells during paraquat (PQ) -induced injury and its underlying mechanisms. The re- search was divided into two parts. The first part of the research was divided into normal control group, PQ group, UTI+PQ group, UTI control group. The second part of the research was divided into negative virus group (including control group, PQ group, PQU+TI group, UTI group) and P-gp siRNA group (including control group, PQ group, PQU+TI group, UTI group) . Negative virus group: the cells were transfected into the blank virus; siRNA P-gp group: the cells were transfected with P-gp siRNA virus. HK-2 cells were routinely cultured. After 800 μmol/L PQ treatment, the changes of P-gp protein levels in the HK-2 cells were determined by West-ern-blot (WB) . Then, transfected lentivirus bringing P-gp silent gene, the cell viability was determined by CCK-8 assay, the expression of P-gp in the cells after transfection was detected by WB and the concentration of PQ in HK-2 cells were measured by high performance liquid chromatography (HPLC) . Compared with the normal control group, the P-gp expression of PQ group had no significantly changes (>0.05) . Compared with the PQ group, the P-gp expression of UTI+PQ group significantly increased (>0.05) . Compared with the corre-sponding control siRNA group, the P-gp siRNA group had no significantly changes in cell viability (>0.05) . and significantly decreased in P-gp expression. Compared with the corresponding control siRNA group, the P-gp siRNA group had no significantly changes in PQ concentration in HK-2 cell (>0.05) , but compared with P-gp siRNA PQ group, the PQ concentration of P-gp siRNA PQ+UTI group significantly decrease (<0.05) . UTI significantly reduced the accumulation of PQ in HK-2 cells and increased the viability of HK-2 cells in vitro may be not by increased P-gp activity. UTI could significantly reduce HK-2 cell injury induced by PQ in vitro and improve the survival rate of HK-2 cells. It may not be related to the up regulation of P-gp expres-sion.
探讨乌司他丁（UTI）上调P-糖蛋白对百草枯（PQ）诱导的HK-2细胞损伤的保护作用及其潜在机制。研究分为两部分。第一部分研究分为正常对照组、PQ组、UTI+PQ组、UTI对照组。第二部分研究分为阴性病毒组（包括对照组、PQ组、PQU+TI组、UTI组）和P-gp siRNA组（包括对照组、PQ组、PQU+TI组、UTI组）。阴性病毒组：细胞转染空白病毒；siRNA P-gp组：细胞转染P-gp siRNA病毒。常规培养HK-2细胞。经800μmol/L PQ处理后，采用蛋白质免疫印迹法（WB）检测HK-2细胞中P-糖蛋白水平的变化。然后，转染携带P-gp沉默基因的慢病毒，采用CCK-8法检测细胞活力，通过WB检测转染后细胞中P-糖蛋白的表达，采用高效液相色谱法（HPLC）测定HK-2细胞中PQ的浓度。与正常对照组比较，PQ组P-糖蛋白表达无明显变化（>0.05）。与PQ组比较，UTI+PQ组P-糖蛋白表达明显增加（>0.05）。与相应的对照siRNA组比较，P-gp siRNA组细胞活力无明显变化（>0.05），P-糖蛋白表达明显降低。与相应的对照siRNA组比较，P-gp siRNA组HK-2细胞中PQ浓度无明显变化（>0.05），但与P-gp siRNA PQ组比较，P-gp siRNA PQ+UTI组PQ浓度明显降低（<0.05）。UTI可显著降低PQ在HK-2细胞中的蓄积，体外增加HK-2细胞活力可能并非通过增加P-糖蛋白活性。UTI可显著减轻体外PQ诱导的HK-2细胞损伤，提高HK-2细胞存活率。这可能与上调P-糖蛋白表达无关。
