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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: 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.

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

PQ induced ROS generation in RAW264.7 cells. Cells were treated with various concentration of PQ for 24 h. After treatment, the cells were incubated with 10 μM carboxy-H2DCF-DA for 60 min, and the intracellular ROS levels were measured by flow cytometry. The fluorescence intensity is proportional to the amount of ROS produced by the cells. Data represent the mean ± SD of at least three independent experiments. ***p<0.001 indicates a significant difference as compared to the control group.
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f4-bt-23-407: PQ induced ROS generation in RAW264.7 cells. Cells were treated with various concentration of PQ for 24 h. After treatment, the cells were incubated with 10 μM carboxy-H2DCF-DA for 60 min, and the intracellular ROS levels were measured by flow cytometry. The fluorescence intensity is proportional to the amount of ROS produced by the cells. Data represent the mean ± SD of at least three independent experiments. ***p<0.001 indicates a significant difference as compared to the control group.

Mentions: To investigate whether ROS mediate PQ-induced apoptosis, we measured the intracellular ROS levels after PQ treatment. Because PQ is known to induce ROS formation through the activation of enzymes initiating redox cycling of the herbicide, it would not be unexpected that ROS generation mediates apoptosis (Ali et al., 1996; Castello et al., 2007; Mussi and Calcaterra, 2010). We used carboxy-H2DCF-DA, a ROS-sensitive fluorometric probe, to measure ROS production by flow cytometry. As shown in Fig. 4, PQ treatment significantly enhanced ROS generation in RAW264.7 cells.


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)

PQ induced ROS generation in RAW264.7 cells. Cells were treated with various concentration of PQ for 24 h. After treatment, the cells were incubated with 10 μM carboxy-H2DCF-DA for 60 min, and the intracellular ROS levels were measured by flow cytometry. The fluorescence intensity is proportional to the amount of ROS produced by the cells. Data represent the mean ± SD of at least three independent experiments. ***p<0.001 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

f4-bt-23-407: PQ induced ROS generation in RAW264.7 cells. Cells were treated with various concentration of PQ for 24 h. After treatment, the cells were incubated with 10 μM carboxy-H2DCF-DA for 60 min, and the intracellular ROS levels were measured by flow cytometry. The fluorescence intensity is proportional to the amount of ROS produced by the cells. Data represent the mean ± SD of at least three independent experiments. ***p<0.001 indicates a significant difference as compared to the control group.
Mentions: To investigate whether ROS mediate PQ-induced apoptosis, we measured the intracellular ROS levels after PQ treatment. Because PQ is known to induce ROS formation through the activation of enzymes initiating redox cycling of the herbicide, it would not be unexpected that ROS generation mediates apoptosis (Ali et al., 1996; Castello et al., 2007; Mussi and Calcaterra, 2010). We used carboxy-H2DCF-DA, a ROS-sensitive fluorometric probe, to measure ROS production by flow cytometry. As shown in Fig. 4, PQ treatment significantly enhanced ROS generation in RAW264.7 cells.

Bottom Line: 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.

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