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Paraquat Induces Apoptosis through a Mitochondria-Dependent Pathway in RAW264.7 Cells.

Jang YJ, Won JH, Back MJ, Fu Z, Jang JM, Ha HC, Hong S, Chang M, Kim DK - Biomol Ther (Seoul) (2015)

Bottom Line: However, the effect of PQ on macrophage cell viability remains unclear.In conclusion, PQ induces apoptosis in RAW264.7 cells through a ROS-mediated mitochondrial pathway.Thus, our study improves our knowledge of PQ-induced toxicity, and may give us a greater understanding of how PQ affects the immune system.

View Article: PubMed Central - PubMed

Affiliation: Department of Health, Social, and Clinical Pharmacy, College of Pharmacy, Chung-Ang University, Seoul 156-756, Republic of Korea.

ABSTRACT
Paraquat dichloride (N,N-dimethyl-4-4'-bipiridinium, PQ) is an extremely toxic chemical that is widely used in herbicides. PQ generates reactive oxygen species (ROS) and causes multiple organ failure. In particular, PQ has been reported to be an immunotoxic agrochemical compound. PQ was shown to decrease the number of macrophages in rats and suppress monocyte phagocytic activity in mice. However, the effect of PQ on macrophage cell viability remains unclear. In this study, we evaluated the cytotoxic effect of PQ on the mouse macrophage cell line, RAW264.7 and its possible mechanism of action. RAW264.7 cells were treated with PQ (0, 75, and 150 μM), and cellular apoptosis, mitochondrial membrane potential (MMP), and intracellular ROS levels were determined. Morphological changes to the cell nucleus and cellular apoptosis were also evaluated by DAPI and Annexin V staining, respectively. In this study, PQ induced apoptotic cell death by dose-dependently decreasing MMP. Additionally, PQ increased the cleaved form of caspase-3, an apoptotic marker. In conclusion, PQ induces apoptosis in RAW264.7 cells through a ROS-mediated mitochondrial pathway. Thus, our study improves our knowledge of PQ-induced toxicity, and may give us a greater understanding of how PQ affects the immune system.

No MeSH data available.


Related in: MedlinePlus

Effect of PQ on RAW264.7 cell viability. Cell viability was assessed using the CCK method. Cells were treated with various concentrations of PQ for 24 h, followed by the addition of 10 μL CCK to each well. Following 1 h incubation at 37°C, absorbance was measured at 450 nm with microplate reader. The cell viability was expressed as the optical density percentage of the treatment versus the control. **p<0.01 indicates a significant difference as compared to the control group.
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f1-bt-23-407: Effect of PQ on RAW264.7 cell viability. Cell viability was assessed using the CCK method. Cells were treated with various concentrations of PQ for 24 h, followed by the addition of 10 μL CCK to each well. Following 1 h incubation at 37°C, absorbance was measured at 450 nm with microplate reader. The cell viability was expressed as the optical density percentage of the treatment versus the control. **p<0.01 indicates a significant difference as compared to the control group.

Mentions: To detect the effects of PQ on RAW264.7 cell viability, the cells were incubated with PQ (0–200 μM) for 24 h (Styles, 1974; Cappelletti et al., 1998; Mitsopoulos and Suntres, 2010, 2011; Cheng et al., 2012; Kang et al., 2013; Wang et al., 2014). After incubation, cell viability was evaluated using a CCK-8 assay. As shown in Fig. 1, PQ dose-dependently decreased cell viability. Treatment of RAW264.7 cells with 150 μM PQ for 24 h reduced cell viability approximately 50%.


Paraquat Induces Apoptosis through a Mitochondria-Dependent Pathway in RAW264.7 Cells.

Jang YJ, Won JH, Back MJ, Fu Z, Jang JM, Ha HC, Hong S, Chang M, Kim DK - Biomol Ther (Seoul) (2015)

Effect of PQ on RAW264.7 cell viability. Cell viability was assessed using the CCK method. Cells were treated with various concentrations of PQ for 24 h, followed by the addition of 10 μL CCK to each well. Following 1 h incubation at 37°C, absorbance was measured at 450 nm with microplate reader. The cell viability was expressed as the optical density percentage of the treatment versus the control. **p<0.01 indicates a significant difference as compared to the control group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556199&req=5

f1-bt-23-407: Effect of PQ on RAW264.7 cell viability. Cell viability was assessed using the CCK method. Cells were treated with various concentrations of PQ for 24 h, followed by the addition of 10 μL CCK to each well. Following 1 h incubation at 37°C, absorbance was measured at 450 nm with microplate reader. The cell viability was expressed as the optical density percentage of the treatment versus the control. **p<0.01 indicates a significant difference as compared to the control group.
Mentions: To detect the effects of PQ on RAW264.7 cell viability, the cells were incubated with PQ (0–200 μM) for 24 h (Styles, 1974; Cappelletti et al., 1998; Mitsopoulos and Suntres, 2010, 2011; Cheng et al., 2012; Kang et al., 2013; Wang et al., 2014). After incubation, cell viability was evaluated using a CCK-8 assay. As shown in Fig. 1, PQ dose-dependently decreased cell viability. Treatment of RAW264.7 cells with 150 μM PQ for 24 h reduced cell viability approximately 50%.

Bottom Line: However, the effect of PQ on macrophage cell viability remains unclear.In conclusion, PQ induces apoptosis in RAW264.7 cells through a ROS-mediated mitochondrial pathway.Thus, our study improves our knowledge of PQ-induced toxicity, and may give us a greater understanding of how PQ affects the immune system.

View Article: PubMed Central - PubMed

Affiliation: Department of Health, Social, and Clinical Pharmacy, College of Pharmacy, Chung-Ang University, Seoul 156-756, Republic of Korea.

ABSTRACT
Paraquat dichloride (N,N-dimethyl-4-4'-bipiridinium, PQ) is an extremely toxic chemical that is widely used in herbicides. PQ generates reactive oxygen species (ROS) and causes multiple organ failure. In particular, PQ has been reported to be an immunotoxic agrochemical compound. PQ was shown to decrease the number of macrophages in rats and suppress monocyte phagocytic activity in mice. However, the effect of PQ on macrophage cell viability remains unclear. In this study, we evaluated the cytotoxic effect of PQ on the mouse macrophage cell line, RAW264.7 and its possible mechanism of action. RAW264.7 cells were treated with PQ (0, 75, and 150 μM), and cellular apoptosis, mitochondrial membrane potential (MMP), and intracellular ROS levels were determined. Morphological changes to the cell nucleus and cellular apoptosis were also evaluated by DAPI and Annexin V staining, respectively. In this study, PQ induced apoptotic cell death by dose-dependently decreasing MMP. Additionally, PQ increased the cleaved form of caspase-3, an apoptotic marker. In conclusion, PQ induces apoptosis in RAW264.7 cells through a ROS-mediated mitochondrial pathway. Thus, our study improves our knowledge of PQ-induced toxicity, and may give us a greater understanding of how PQ affects the immune system.

No MeSH data available.


Related in: MedlinePlus