Glyphosate Commercial Formulation Causes Cytotoxicity, Oxidative Effects, and Apoptosis on Human Cells

Abstract

In the present study, the effects on oxidative balance and cellular end points of glyphosate, aminomethylphosphonic acid (AMPA), and a glyphosate formulation (G formulation) were examined in HepG2 cell line, at dilution levels far below agricultural recommendations. Our results show that G formulation had toxic effects while no effects were found with acid glyphosate and AMPA treatments. Glyphosate formulation exposure produced an increase in reactive oxygen species, nitrotyrosine formation, superoxide dismutase activity, and glutathione (GSH) levels, while no effects were observed for catalase and GSH-S-transferase activities. Also, G formulation triggered caspase 3/7 activation and hence induced apoptosis pathway in this cell line. Aminomethylphosphonic acid exposure produced an increase in GSH levels while no differences were observed in other antioxidant parameters. No effects were observed when the cells were exposed to acid glyphosate. These results confirm that G formulations have adjuvants working together with the active ingredient and causing toxic effects that are not seen with acid glyphosate.

Influence of the spray adjuvant on the toxicity effects of a glyphosate formulation

“…We found an important increase in ROS production (139% and 116% of control, p < 0.05) in cells treated with Atanor Ò and mixture respectively. These results agree with our previous findings and others (Chaufan et al, 2014;Heu et al, 2012) where ROS formation in different type of cells was measured. All together these findings demonstrated that Impacto Ò and Atanor Ò cause changes to redox state and ROS production as shown by studies (Heu et al, 2012;Gehin et al, 2006;Li, 2008;Okai et al, 2004;Gong and Han, 2006).…”

“…No differences between control and treatment groups were observed for SOD and GST activities in HEp-2 cells. This result agrees with our previous findings that observed an increase in GSH levels when cells were treated with other glyphosate formulation (Chaufan et al, 2014).…”

“…In addition to the increase in ROS, we observed high levels in caspase 3/7 activity with all the agrochemicals used. In this area we demonstrated that cell death induced by glyphosate formulation is predominantly apoptotic, as determined by DAPI staining and caspase-3/7 activation (Chaufan et al, 2014). However, the addition of adjuvant to the glyphosate formulation did not increase morphological changes (condensed and fragmented nuclei) consistent with apoptotic cell death.…”

Influence of the spray adjuvant on the toxicity effects of a glyphosate formulation

“…We found an important increase in ROS production (139% and 116% of control, p < 0.05) in cells treated with Atanor Ò and mixture respectively. These results agree with our previous findings and others (Chaufan et al, 2014;Heu et al, 2012) where ROS formation in different type of cells was measured. All together these findings demonstrated that Impacto Ò and Atanor Ò cause changes to redox state and ROS production as shown by studies (Heu et al, 2012;Gehin et al, 2006;Li, 2008;Okai et al, 2004;Gong and Han, 2006).…”

“…No differences between control and treatment groups were observed for SOD and GST activities in HEp-2 cells. This result agrees with our previous findings that observed an increase in GSH levels when cells were treated with other glyphosate formulation (Chaufan et al, 2014).…”

“…In addition to the increase in ROS, we observed high levels in caspase 3/7 activity with all the agrochemicals used. In this area we demonstrated that cell death induced by glyphosate formulation is predominantly apoptotic, as determined by DAPI staining and caspase-3/7 activation (Chaufan et al, 2014). However, the addition of adjuvant to the glyphosate formulation did not increase morphological changes (condensed and fragmented nuclei) consistent with apoptotic cell death.…”

Acute exposition to Roundup Transorb® induces systemic oxidative stress and alterations in the expression of newly sequenced genes in silverside fish (Odontesthes humensis)

“…The increased DFI observed in the fish exposed to RDT in the present study might be an indirect consequence of the oxidative stress induced by lipid peroxidation, as mentioned before, or a direct result of excess ROS. We also observed high levels of apoptosis in the fish exposed to RDT, which may have occurred because of changes in the plasma membrane or generation of excess ROS, as these play critical roles in signaling pathways that induce apoptosis (Chaufan et al 2014). Certain previous studies have reported similar results, showing an increase in the level of cellular apoptosis in animals exposed to herbicides (Topal et al 2015;Sulukan et al 2017).…”

Micronucleus Formation Induced by Glyphosate and Glyphosate-Based Herbicides in Human Peripheral White Blood Cells

“…According to Wozniak et al (30), genotoxicity detected in HMWB cells after exposure to technical glyphosate, Roundup 360 PLUS or the metabolite of glyphosate (aminomethylphosphonic acid, AMPA) was not the result of direct interaction of these compounds with the genetic material because no DNA adducts have been formed, rather due to effects mediated by reactive oxygen species (ROS) induced by the chemicals, which may also explain increased cell death. There is no clear evidence on the ROS-inducing potential of glyphosate (22,25,52); however, GBHs have been shown to induce oxidative stress (46,53,54), supporting the role of co-formulants in the cytotoxic effects observed in our experiments.…”

Influence of the spray adjuvant on the toxicity effects of a glyphosate formulation

“…We found an important increase in ROS production (139% and 116% of control, p < 0.05) in cells treated with Atanor Ò and mixture respectively. These results agree with our previous findings and others (Chaufan et al, 2014;Heu et al, 2012) where ROS formation in different type of cells was measured. All together these findings demonstrated that Impacto Ò and Atanor Ò cause changes to redox state and ROS production as shown by studies (Heu et al, 2012;Gehin et al, 2006;Li, 2008;Okai et al, 2004;Gong and Han, 2006).…”

“…No differences between control and treatment groups were observed for SOD and GST activities in HEp-2 cells. This result agrees with our previous findings that observed an increase in GSH levels when cells were treated with other glyphosate formulation (Chaufan et al, 2014).…”

“…In addition to the increase in ROS, we observed high levels in caspase 3/7 activity with all the agrochemicals used. In this area we demonstrated that cell death induced by glyphosate formulation is predominantly apoptotic, as determined by DAPI staining and caspase-3/7 activation (Chaufan et al, 2014). However, the addition of adjuvant to the glyphosate formulation did not increase morphological changes (condensed and fragmented nuclei) consistent with apoptotic cell death.…”

Acute exposition to Roundup Transorb® induces systemic oxidative stress and alterations in the expression of newly sequenced genes in silverside fish (Odontesthes humensis)

“…The increased DFI observed in the fish exposed to RDT in the present study might be an indirect consequence of the oxidative stress induced by lipid peroxidation, as mentioned before, or a direct result of excess ROS. We also observed high levels of apoptosis in the fish exposed to RDT, which may have occurred because of changes in the plasma membrane or generation of excess ROS, as these play critical roles in signaling pathways that induce apoptosis (Chaufan et al 2014). Certain previous studies have reported similar results, showing an increase in the level of cellular apoptosis in animals exposed to herbicides (Topal et al 2015;Sulukan et al 2017).…”

Micronucleus Formation Induced by Glyphosate and Glyphosate-Based Herbicides in Human Peripheral White Blood Cells

“…According to Wozniak et al (30), genotoxicity detected in HMWB cells after exposure to technical glyphosate, Roundup 360 PLUS or the metabolite of glyphosate (aminomethylphosphonic acid, AMPA) was not the result of direct interaction of these compounds with the genetic material because no DNA adducts have been formed, rather due to effects mediated by reactive oxygen species (ROS) induced by the chemicals, which may also explain increased cell death. There is no clear evidence on the ROS-inducing potential of glyphosate (22,25,52); however, GBHs have been shown to induce oxidative stress (46,53,54), supporting the role of co-formulants in the cytotoxic effects observed in our experiments.…”

Influence of the spray adjuvant on the toxicity effects of a glyphosate formulation

“…We found an important increase in ROS production (139% and 116% of control, p < 0.05) in cells treated with Atanor Ò and mixture respectively. These results agree with our previous findings and others (Chaufan et al, 2014;Heu et al, 2012) where ROS formation in different type of cells was measured. All together these findings demonstrated that Impacto Ò and Atanor Ò cause changes to redox state and ROS production as shown by studies (Heu et al, 2012;Gehin et al, 2006;Li, 2008;Okai et al, 2004;Gong and Han, 2006).…”

“…No differences between control and treatment groups were observed for SOD and GST activities in HEp-2 cells. This result agrees with our previous findings that observed an increase in GSH levels when cells were treated with other glyphosate formulation (Chaufan et al, 2014).…”

“…In addition to the increase in ROS, we observed high levels in caspase 3/7 activity with all the agrochemicals used. In this area we demonstrated that cell death induced by glyphosate formulation is predominantly apoptotic, as determined by DAPI staining and caspase-3/7 activation (Chaufan et al, 2014). However, the addition of adjuvant to the glyphosate formulation did not increase morphological changes (condensed and fragmented nuclei) consistent with apoptotic cell death.…”

Acute exposition to Roundup Transorb® induces systemic oxidative stress and alterations in the expression of newly sequenced genes in silverside fish (Odontesthes humensis)

“…The increased DFI observed in the fish exposed to RDT in the present study might be an indirect consequence of the oxidative stress induced by lipid peroxidation, as mentioned before, or a direct result of excess ROS. We also observed high levels of apoptosis in the fish exposed to RDT, which may have occurred because of changes in the plasma membrane or generation of excess ROS, as these play critical roles in signaling pathways that induce apoptosis (Chaufan et al 2014). Certain previous studies have reported similar results, showing an increase in the level of cellular apoptosis in animals exposed to herbicides (Topal et al 2015;Sulukan et al 2017).…”

Micronucleus Formation Induced by Glyphosate and Glyphosate-Based Herbicides in Human Peripheral White Blood Cells

“…According to Wozniak et al (30), genotoxicity detected in HMWB cells after exposure to technical glyphosate, Roundup 360 PLUS or the metabolite of glyphosate (aminomethylphosphonic acid, AMPA) was not the result of direct interaction of these compounds with the genetic material because no DNA adducts have been formed, rather due to effects mediated by reactive oxygen species (ROS) induced by the chemicals, which may also explain increased cell death. There is no clear evidence on the ROS-inducing potential of glyphosate (22,25,52); however, GBHs have been shown to induce oxidative stress (46,53,54), supporting the role of co-formulants in the cytotoxic effects observed in our experiments.…”