[乌司他丁对百草枯诱导的HK-2细胞损伤的保护作用及潜在机制]
[The protective effect of ulinastatin on paraquat-induced injury in HK-2 cells and the underlying mechanisms].
作者信息
She Xingrong, Hong Guangliang, Tan Jiaping, Zhao Guangju, Li Mengfang, Lu Zhongqiu
机构信息
Emergency Department of the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
Emergency Department of the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China; E-mial:
出版信息
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2015 Jul;33(7):501-6.
OBJECTIVE
To investigate the protective effect of ulinastatin (UTI) on HK-2 cells during paraquat (PQ)-induced injury and its underlying mechanisms.
METHODS
Routinely cultured HK-2 cells were divided into blank control group, PQ group, UTI+PQ group and UTI group. Cell viability was determined by CCK-8 assay. The concentration of PQ in HK-2 cells were measured by high performance liquid chromatography (HPLC). The production of total reactive oxygen species (ROS) were detected by fluorescence microscopy. The activities of superoxide dismutase activity (SOD) and the content of malondialdehyde (MDA) in HK-2 cells were observed by chemical colorimetry. The levels of tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) were measured by enzyme-linked immunosorbent assay (ELISA).
RESULTS
PQ, even at a dose of 200 µM, could significant suppress the viability of HK-2 cells in a dose-dependent and time-dependent. UTI showed no significant inhibitory effect on the viability of HK-2 cells when given at a dose below 8 000 U/ml (P > 0.05). Compared with the PQ group, the UTI+PQ group had significantly increased the viability of HK-2 cells in a dose-dependent of UTI (P < 0.05). Compared with the PQ group on the same hour, the UTI+PQ group showed decreased in PQ concentration in HK-2 cells (P < 0.05 for all except 6 h). Compared with the blank control group, the PQ group had significantly decreased SOD activity and significantly increased ROS level and MDA content (P < 0.05). Compared with the PQ group, the UTI+PQ group had significantly increased SOD activity and significantly decreased ROS level and MDA content (P < 0.05). Compared with the blank control group, the PQ group had significantly increased IL-6 and TNF-α level (P < 0.05); Compared with the PQ group, the UTI+PQ group had significantly decreased IL-6 and TNF-α level (P < 0.05).
CONCLUSION
UTI significantly reduces the PQ-induced oxidative damage and inflammatory injury and its mechanism may be by reducing the accumulation of PQ in HK-2 cells.
目的
探讨乌司他丁（UTI）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。
方法
将常规培养的HK-2细胞分为空白对照组、PQ组、UTI+PQ组和UTI组。采用CCK-8法检测细胞活力。用高效液相色谱法（HPLC）测定HK-2细胞中PQ的浓度。通过荧光显微镜检测总活性氧（ROS）的产生。采用化学比色法观察HK-2细胞中超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量。用酶联免疫吸附测定法（ELISA）检测肿瘤坏死因子α（TNF-α）和白细胞介素6（IL-6）水平。
结果
即使剂量为200μM，PQ也能以剂量和时间依赖性方式显著抑制HK-2细胞的活力。当剂量低于8000U/ml时，UTI对HK-2细胞活力无显著抑制作用（P>0.05）。与PQ组相比，UTI+PQ组HK-2细胞活力随UTI剂量增加而显著升高（P<0.05）。与同一时间的PQ组相比，UTI+PQ组HK-2细胞中PQ浓度降低（除6小时外，其余均P<0.05）。与空白对照组相比，PQ组SOD活性显著降低，ROS水平和MDA含量显著升高（P<0.05）。与PQ组相比，UTI+PQ组SOD活性显著升高，ROS水平和MDA含量显著降低（P<0.05）。与空白对照组相比，PQ组IL-6和TNF-α水平显著升高（P<0.05）；与PQ组相比，UTI+PQ组IL-6和TNF-α水平显著降低（P<0.05）。
结论
UTI可显著减轻PQ诱导的氧化损伤和炎症损伤，其机制可能是减少PQ在HK-2细胞中的蓄积。
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