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Effect of ulinastatin on paraquat-induced-oxidative stress in human type II alveolar epithelial cells.

Meng XX, Wang RL, Gao S, Xie H, Tan JT, Qian YB - World J Emerg Med (2013)

Bottom Line: The survival rate of A549 cells treated with different concentrations of paraquat decreased in a concentration-dependent manner.Whereas there was no decrease in the survival rate of cells treated with 0-4 000 U/mL ulinastatin.The levels of MDA, MPO, and ROS were significantly higher in the paraquat group than in the normal control group after 24-hour-exposure.

View Article: PubMed Central - PubMed

Affiliation: Department of Critical Care Medicine, Shanghai First People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 201620, China.

ABSTRACT

Background: Ulinastatin (UTI) is a urinary trypsin inhibitor extracted and purified from urine of males. This study aimed to explore the effects of UTI on paraquat-induced-oxidative stress in human type II alveolar epithelial cells.

Methods: The human type II alveolar epithelial cells, A549 cells, were cultured in vitro. The A549 cells were treated with different concentrations of paraquat (200, 400, 600, 800, 1 000, 1 200 µmol/L) and ulinastatin(0, 2 000, 4 000, 6 000, 8 000 U/mL) for 24 hours, the cell viability was measured by cell counting kit-8 and the median lethal concentration was selected. In order to establish an in vitro model of paraquat intoxication and to determine the safe dose of ulinastatin, we calculated LD50 using cell counting kit-8 to determine the survival rate of the cells. A549 cells were divided into normal control group, paraquat group and paraquat+ulinastatin group. The levels of malondialdehyde (MDA) and myeloperoxidase (MPO) were detected by biochemistry colorimetry, while the level of reactive oxygen spies (ROS) was detected by DCFH-DA assay.

Results: The survival rate of A549 cells treated with different concentrations of paraquat decreased in a concentration-dependent manner. Whereas there was no decrease in the survival rate of cells treated with 0-4 000 U/mL ulinastatin. The levels of MDA, MPO, and ROS were significantly higher in the paraquat group than in the normal control group after 24-hour-exposure. And the survival rate of the paraquat+ulinastatin group was higher than that of the paraquat group, but lower than that of the normal control group. The levels of MDA, MPO, and ROS were lower than those of the paraquat group.

Conclusion: Ulinastatin can alleviate the paraquat-induced A549 cell damage by reducing oxidative stress.

No MeSH data available.


Related in: MedlinePlus

Effect of different PQ concentrations on the cell survival rate. The survival rate of the cells decreased as the concentration increased. Compared to the control group, *P<0.05; compared to the control group, **P<0.01.
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Figure 1: Effect of different PQ concentrations on the cell survival rate. The survival rate of the cells decreased as the concentration increased. Compared to the control group, *P<0.05; compared to the control group, **P<0.01.

Mentions: After incubation for 24 hours in PQ solutions with concentrations of 200, 400, 600, 800, 1 000 and 1 200 µmol/L, the survival rate of the cells decreased as the concentration increased. As indicated by Figure 1, the survival rate in groups of 400, 600, 800, 1 000, 1 200 µmol/L were significantly different from the control group (P<0.05). For the purpose of this experiment, we artificially set the PQ concentration of 800 µmol/L where the survival rate was 57.01%, as the intervention point. At the UTI concentration of 2 000 and 4 000 U/mL, the cell survival rates were 94.9%±4.3%, 91.1%±1.6% respectively. This indicated there was no damage to the cells at these concentrations, whereas the survival rate decreased significantly at the concentrations of 6 000 and 8 000 U/mL (75.6%±3.2% and 56.5%±1.1% respectively). Thus we chose 2 000 and 4 000 U/mL as the operating concentration in our study.


Effect of ulinastatin on paraquat-induced-oxidative stress in human type II alveolar epithelial cells.

Meng XX, Wang RL, Gao S, Xie H, Tan JT, Qian YB - World J Emerg Med (2013)

Effect of different PQ concentrations on the cell survival rate. The survival rate of the cells decreased as the concentration increased. Compared to the control group, *P<0.05; compared to the control group, **P<0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4129841&req=5

Figure 1: Effect of different PQ concentrations on the cell survival rate. The survival rate of the cells decreased as the concentration increased. Compared to the control group, *P<0.05; compared to the control group, **P<0.01.
Mentions: After incubation for 24 hours in PQ solutions with concentrations of 200, 400, 600, 800, 1 000 and 1 200 µmol/L, the survival rate of the cells decreased as the concentration increased. As indicated by Figure 1, the survival rate in groups of 400, 600, 800, 1 000, 1 200 µmol/L were significantly different from the control group (P<0.05). For the purpose of this experiment, we artificially set the PQ concentration of 800 µmol/L where the survival rate was 57.01%, as the intervention point. At the UTI concentration of 2 000 and 4 000 U/mL, the cell survival rates were 94.9%±4.3%, 91.1%±1.6% respectively. This indicated there was no damage to the cells at these concentrations, whereas the survival rate decreased significantly at the concentrations of 6 000 and 8 000 U/mL (75.6%±3.2% and 56.5%±1.1% respectively). Thus we chose 2 000 and 4 000 U/mL as the operating concentration in our study.

Bottom Line: The survival rate of A549 cells treated with different concentrations of paraquat decreased in a concentration-dependent manner.Whereas there was no decrease in the survival rate of cells treated with 0-4 000 U/mL ulinastatin.The levels of MDA, MPO, and ROS were significantly higher in the paraquat group than in the normal control group after 24-hour-exposure.

View Article: PubMed Central - PubMed

Affiliation: Department of Critical Care Medicine, Shanghai First People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 201620, China.

ABSTRACT

Background: Ulinastatin (UTI) is a urinary trypsin inhibitor extracted and purified from urine of males. This study aimed to explore the effects of UTI on paraquat-induced-oxidative stress in human type II alveolar epithelial cells.

Methods: The human type II alveolar epithelial cells, A549 cells, were cultured in vitro. The A549 cells were treated with different concentrations of paraquat (200, 400, 600, 800, 1 000, 1 200 µmol/L) and ulinastatin(0, 2 000, 4 000, 6 000, 8 000 U/mL) for 24 hours, the cell viability was measured by cell counting kit-8 and the median lethal concentration was selected. In order to establish an in vitro model of paraquat intoxication and to determine the safe dose of ulinastatin, we calculated LD50 using cell counting kit-8 to determine the survival rate of the cells. A549 cells were divided into normal control group, paraquat group and paraquat+ulinastatin group. The levels of malondialdehyde (MDA) and myeloperoxidase (MPO) were detected by biochemistry colorimetry, while the level of reactive oxygen spies (ROS) was detected by DCFH-DA assay.

Results: The survival rate of A549 cells treated with different concentrations of paraquat decreased in a concentration-dependent manner. Whereas there was no decrease in the survival rate of cells treated with 0-4 000 U/mL ulinastatin. The levels of MDA, MPO, and ROS were significantly higher in the paraquat group than in the normal control group after 24-hour-exposure. And the survival rate of the paraquat+ulinastatin group was higher than that of the paraquat group, but lower than that of the normal control group. The levels of MDA, MPO, and ROS were lower than those of the paraquat group.

Conclusion: Ulinastatin can alleviate the paraquat-induced A549 cell damage by reducing oxidative stress.

No MeSH data available.


Related in: MedlinePlus