- Email: [email protected]
AKT signaling pathways
S32
D. Rossin et al. / Free Radical Biology and Medicine 108 (2017) S18–S107
3
Center of Applied Thai Traditional Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand
Keywords: Antioxidant defense; Melanogenesis; Nuclear factor E2-related factor 2 (Nrf2); Polyherbal formula; Ultraviolet A
Thai herbal antipyretic 22 formula (APF22), a polyherbal formula, has been used to treat dermatologic problems including hyperpigmentation. Exposure to ultraviolet A (UVA) causes abnormal melanin production induced by photooxidative stress in the skin. Our previous study suggests that pharmacological actions of the APF22 may be attributed to gallic acid (GA), a possible active ingredient of this formula. Aims: This study investigated the protective effects of the APF22 extracts, ferulic acid (FA) and GA (used as reference compounds), on melanogenesis through modulation of Nrf2 signaling and promotion of antioxidant defenses in mouse melanoma (B16F10) cells exposed to UVA (8 J/cm2). Results: The APF22 extracts, FA and GA provided anti-melanogenic effect on UVA-irradiated B16F10 cells by suppression of melanin synthesis as well as tyrosinase activity and protein levels. Moreover, APF22 extracts, FA and GA were able to activate Nrf2-ARE signaling and induce mRNA levels of antioxidant defenses including glutathione (GSH), catalase (CAT), glutathione peroxidase (GPx) and the glutathione-s-transferase (GST) in UVA-irradiated cells. Our study concluded that APF22 extracts suppressed UVAmediated melanogenesis possibly via redox mechanisms involving activation of Nrf2-ARE signaling. Moreover, pharmacological action of the APF22 extracts may be attributed to FA and GA.
viability in LtP with IC50 values in the low mM range, while IC50 values for J774 macrophages were considerably higher. A similar structure without EP bridge (1,4-cineole) resulted in no detectable radicals and less cytotoxicity in LtP. The IC50 values for LtP viability in the presence of Asc or Art were increased by the spin trap DMPO. ICP-OES measurements revealed that in LtP the total iron concentration was twice as high as values in J774 macrophages. Studies of the labile iron pool (LIP) in LtP by low temperature ESR revealed an oxidation of the LIP by Asc. These data demonstrate that radical formation from Asc/Art in LtP is an essential part of their antileishmanial mechanism. E-mail address: [email protected] (L. Gille) Acknowledgements
Funding was provided by Austrian Science Fund (FWF) grant P27814-B22, Austrian Exchange Office (OEAD). http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.130
P-046
Cordycepin induced MA-10 mouse Leydig tumor cell apoptosis by regulating p38 MAPKs and PI3K/AKT signaling pathways Bu-Miin Huang National Cheng Kung University, Taiwan, Republic of China
E-mail address: [email protected] (U. Panich) http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.129
P-045
The activation of the endoperoxide ascaridole in Leishmania Lars Gille 1, Gerald Geroldinger 1, Matthias Tonner 1, Hubert Hettegger 2, Markus Bacher 2, Lianet Monzote 3, Martin Walter 4, Katrin Staniek 1, Thomas Rosenau 2 1
Univ. of Veterinary Medicine Vienna, Austria Department of Chemistry, University of Natural Resources and Life Sciences, Tulln, Austria 3 Parasitology Department, Institute of Tropical Medicine “Pedro Kouri”, Havana, Cuba 4 Department of Environmental Geosciences, University of Vienna, Vienna, Austria 2
Keywords: Ascaridole; endoperoxides; Leishmania; electron spin resonance; iron
Endoperoxides (EP) are successfully used in malaria therapy. It was shown in a mouse model that the EP ascaridole (Asc) is useful for the treatment of cutaneous leishmaniasis. The current study explored the activation of Asc and artemisinin (Art) in biomimetic systems and inside Leishmania tarentolae promastigotes (LtP) as model for pathogenic Leishmania in comparison to J774 macrophages. Using ESR spin-trapping we identified isopropyl radicals arising from reduction by Fe2+ in cell-free systems. Combined GC/ NMR analysis confirmed the loss of isopropyl residues from Asc. In LtP carbon-centered radicals were identified in the presence of Asc and Art by ESR spin-trapping. Both Asc and Art inhibited the
Keywords: Cordycepin; MA-10 cell; Leydig tumor; apoptosis; caspase; p38; AKT; cell cycle
The p38 MAPKs play important roles in the regulation of balance between cell survival and cell death on the development of various cancers. However, the roles of p38 MAPKs regulating apoptotic effects on Leydig tumor cells remain unclear. In the present study, we showed that cordycepin (3′-deoxyadenosine) selectively induced apoptosis in MA-10 mouse Leydig tumor cells through regulating the p38 MAPK and PI3K/AKT signaling pathways. Cordycepin reduced viability in MA-10 and TM4 cells, but not cause cell death of primary mouse Leydig cells on moderate concentration. Cordycepin increased reactive oxygen species (ROS) levels, which is associated with the induction of apoptosis as characterized by positive Annexin V binding, activation of caspase3, and cleavage of PARP. Inhibition of p38 MAPKs activity by SB203580 significantly prevented cordycepin-induced apoptosis in MA-10 cells. Co-treatment with wortmannin or the autophagy inhibitor 3-methyladenine (3-MA) elevated levels of apoptosis in cordycepin-treated MA-10 cells. Moreover, cordycepin activated p53, p21 and TGFß; and downregulated CDK2. The antitumor activity of cordycepin-treated MA-10 cells was significantly distinct in severe combined immunodeficiency (SCID) mice in vivo. These results suggested that cordycepin is a highly selective treatment to induce MA-10 cells apoptosis via p38 MAPKs signaling. E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.131
D. Rossin et al. / Free Radical Biology and Medicine 108 (2017) S18–S107
3
Center of Applied Thai Traditional Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand
Keywords: Antioxidant defense; Melanogenesis; Nuclear factor E2-related factor 2 (Nrf2); Polyherbal formula; Ultraviolet A
Thai herbal antipyretic 22 formula (APF22), a polyherbal formula, has been used to treat dermatologic problems including hyperpigmentation. Exposure to ultraviolet A (UVA) causes abnormal melanin production induced by photooxidative stress in the skin. Our previous study suggests that pharmacological actions of the APF22 may be attributed to gallic acid (GA), a possible active ingredient of this formula. Aims: This study investigated the protective effects of the APF22 extracts, ferulic acid (FA) and GA (used as reference compounds), on melanogenesis through modulation of Nrf2 signaling and promotion of antioxidant defenses in mouse melanoma (B16F10) cells exposed to UVA (8 J/cm2). Results: The APF22 extracts, FA and GA provided anti-melanogenic effect on UVA-irradiated B16F10 cells by suppression of melanin synthesis as well as tyrosinase activity and protein levels. Moreover, APF22 extracts, FA and GA were able to activate Nrf2-ARE signaling and induce mRNA levels of antioxidant defenses including glutathione (GSH), catalase (CAT), glutathione peroxidase (GPx) and the glutathione-s-transferase (GST) in UVA-irradiated cells. Our study concluded that APF22 extracts suppressed UVAmediated melanogenesis possibly via redox mechanisms involving activation of Nrf2-ARE signaling. Moreover, pharmacological action of the APF22 extracts may be attributed to FA and GA.
viability in LtP with IC50 values in the low mM range, while IC50 values for J774 macrophages were considerably higher. A similar structure without EP bridge (1,4-cineole) resulted in no detectable radicals and less cytotoxicity in LtP. The IC50 values for LtP viability in the presence of Asc or Art were increased by the spin trap DMPO. ICP-OES measurements revealed that in LtP the total iron concentration was twice as high as values in J774 macrophages. Studies of the labile iron pool (LIP) in LtP by low temperature ESR revealed an oxidation of the LIP by Asc. These data demonstrate that radical formation from Asc/Art in LtP is an essential part of their antileishmanial mechanism. E-mail address: [email protected] (L. Gille) Acknowledgements
Funding was provided by Austrian Science Fund (FWF) grant P27814-B22, Austrian Exchange Office (OEAD). http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.130
P-046
Cordycepin induced MA-10 mouse Leydig tumor cell apoptosis by regulating p38 MAPKs and PI3K/AKT signaling pathways Bu-Miin Huang National Cheng Kung University, Taiwan, Republic of China
E-mail address: [email protected] (U. Panich) http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.129
P-045
The activation of the endoperoxide ascaridole in Leishmania Lars Gille 1, Gerald Geroldinger 1, Matthias Tonner 1, Hubert Hettegger 2, Markus Bacher 2, Lianet Monzote 3, Martin Walter 4, Katrin Staniek 1, Thomas Rosenau 2 1
Univ. of Veterinary Medicine Vienna, Austria Department of Chemistry, University of Natural Resources and Life Sciences, Tulln, Austria 3 Parasitology Department, Institute of Tropical Medicine “Pedro Kouri”, Havana, Cuba 4 Department of Environmental Geosciences, University of Vienna, Vienna, Austria 2
Keywords: Ascaridole; endoperoxides; Leishmania; electron spin resonance; iron
Endoperoxides (EP) are successfully used in malaria therapy. It was shown in a mouse model that the EP ascaridole (Asc) is useful for the treatment of cutaneous leishmaniasis. The current study explored the activation of Asc and artemisinin (Art) in biomimetic systems and inside Leishmania tarentolae promastigotes (LtP) as model for pathogenic Leishmania in comparison to J774 macrophages. Using ESR spin-trapping we identified isopropyl radicals arising from reduction by Fe2+ in cell-free systems. Combined GC/ NMR analysis confirmed the loss of isopropyl residues from Asc. In LtP carbon-centered radicals were identified in the presence of Asc and Art by ESR spin-trapping. Both Asc and Art inhibited the
Keywords: Cordycepin; MA-10 cell; Leydig tumor; apoptosis; caspase; p38; AKT; cell cycle
The p38 MAPKs play important roles in the regulation of balance between cell survival and cell death on the development of various cancers. However, the roles of p38 MAPKs regulating apoptotic effects on Leydig tumor cells remain unclear. In the present study, we showed that cordycepin (3′-deoxyadenosine) selectively induced apoptosis in MA-10 mouse Leydig tumor cells through regulating the p38 MAPK and PI3K/AKT signaling pathways. Cordycepin reduced viability in MA-10 and TM4 cells, but not cause cell death of primary mouse Leydig cells on moderate concentration. Cordycepin increased reactive oxygen species (ROS) levels, which is associated with the induction of apoptosis as characterized by positive Annexin V binding, activation of caspase3, and cleavage of PARP. Inhibition of p38 MAPKs activity by SB203580 significantly prevented cordycepin-induced apoptosis in MA-10 cells. Co-treatment with wortmannin or the autophagy inhibitor 3-methyladenine (3-MA) elevated levels of apoptosis in cordycepin-treated MA-10 cells. Moreover, cordycepin activated p53, p21 and TGFß; and downregulated CDK2. The antitumor activity of cordycepin-treated MA-10 cells was significantly distinct in severe combined immunodeficiency (SCID) mice in vivo. These results suggested that cordycepin is a highly selective treatment to induce MA-10 cells apoptosis via p38 MAPKs signaling. E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.131