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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

Effect of PQ on the apoptosis of RAW264.7 cells. Cells were treated with various concentrations of PQ for 24 h, followed by Annexin V-FITC/PI double staining to evaluate apoptosis. The degree of apoptotic cell death was quantified by measuring fluorescence with a FACS CaliburTM flow cytometer. 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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f2-bt-23-407: Effect of PQ on the apoptosis of RAW264.7 cells. Cells were treated with various concentrations of PQ for 24 h, followed by Annexin V-FITC/PI double staining to evaluate apoptosis. The degree of apoptotic cell death was quantified by measuring fluorescence with a FACS CaliburTM flow cytometer. 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 quantify PQ-induced apoptosis, we performed a fluorometric analysis after annexin V-FITC/PI double staining. The rate of apoptosis in control cells was low (5.69%; Fig. 2). However, apoptotic cells, as indicated by Annexin V-FITC positive cells, were dose-dependently increased by PQ (18.27 and 30.55% compared to the control; Fig. 2). To investigate the effects of PQ on the nucleus, cells were stained with DAPI. PQ induced prominent nuclear changes in treated cells (Fig. 3). The nuclei were round, intact, and uniformly stained in control cells; however, PQ exposure induced nuclear condensation, resulting in smaller nuclei that displayed membrane blebbing and fragmentation as the cells died. At 75 and 150 μM PQ, a number of cells exhibited nuclear condensation, membrane blebbing, and apoptotic bodies. Importantly, these aberrant nuclear alterations were not observed in control cells. Thus, PQ induced apoptosis 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)

Effect of PQ on the apoptosis of RAW264.7 cells. Cells were treated with various concentrations of PQ for 24 h, followed by Annexin V-FITC/PI double staining to evaluate apoptosis. The degree of apoptotic cell death was quantified by measuring fluorescence with a FACS CaliburTM flow cytometer. 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

f2-bt-23-407: Effect of PQ on the apoptosis of RAW264.7 cells. Cells were treated with various concentrations of PQ for 24 h, followed by Annexin V-FITC/PI double staining to evaluate apoptosis. The degree of apoptotic cell death was quantified by measuring fluorescence with a FACS CaliburTM flow cytometer. 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 quantify PQ-induced apoptosis, we performed a fluorometric analysis after annexin V-FITC/PI double staining. The rate of apoptosis in control cells was low (5.69%; Fig. 2). However, apoptotic cells, as indicated by Annexin V-FITC positive cells, were dose-dependently increased by PQ (18.27 and 30.55% compared to the control; Fig. 2). To investigate the effects of PQ on the nucleus, cells were stained with DAPI. PQ induced prominent nuclear changes in treated cells (Fig. 3). The nuclei were round, intact, and uniformly stained in control cells; however, PQ exposure induced nuclear condensation, resulting in smaller nuclei that displayed membrane blebbing and fragmentation as the cells died. At 75 and 150 μM PQ, a number of cells exhibited nuclear condensation, membrane blebbing, and apoptotic bodies. Importantly, these aberrant nuclear alterations were not observed in control cells. Thus, PQ induced apoptosis 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