In the present study, we investigated the polarization of monocytes to macrophages, ROS generation during the transition, and mediated protein modifications under the influence of different differentiation inducers. Morphological changes in monocytes in the presence of a differentiation inducer were studied using Confocal Laser Scanning Microscopy (CLSM), and NADPH oxidase expression was studied using Immunoblotting. The regulation of the enzyme complex was studied in both monocytes and macrophages, and the role of EVs secreted from monocytes and their effect on differentiating monocytes as messengers or antioxidant cargo was also studied. Our results showed that monocytes turn into macrophages when they encounter inflammation or differentiation and produce ROS. Polyunsaturated fatty acids (PUFA) are easily damaged by these reactive molecules and can lead to reactive intermediates or products such as aldehydes. Malondialdehyde (MDA), which is one of the final products of polyunsaturated fatty acids peroxidation led to protein modification.
Furthermore, the involvement of EVs derived from monocytes and their co-culture with non-differentiating/differentiating cells showed that EVs through their cargo of regulatory molecules, potentially NOX isotypes contributed to the modulation of polarization of monocytes to distinct macrophage phenotypes. We studied the effect of selected exogenous antioxidant/bioactive compounds isolated from industrial waste to understand their efficacy in protecting the cells against oxidative damage. The intricate equilibrium and transition between the pro-oxidant and antioxidant effects of antioxidants were also investigated.
Anotace v angličtině
In the present study, we investigated the polarization of monocytes to macrophages, ROS generation during the transition, and mediated protein modifications under the influence of different differentiation inducers. Morphological changes in monocytes in the presence of a differentiation inducer were studied using Confocal Laser Scanning Microscopy (CLSM), and NADPH oxidase expression was studied using Immunoblotting. The regulation of the enzyme complex was studied in both monocytes and macrophages, and the role of EVs secreted from monocytes and their effect on differentiating monocytes as messengers or antioxidant cargo was also studied. Our results showed that monocytes turn into macrophages when they encounter inflammation or differentiation and produce ROS. Polyunsaturated fatty acids (PUFA) are easily damaged by these reactive molecules and can lead to reactive intermediates or products such as aldehydes. Malondialdehyde (MDA), which is one of the final products of polyunsaturated fatty acids peroxidation led to protein modification.
Furthermore, the involvement of EVs derived from monocytes and their co-culture with non-differentiating/differentiating cells showed that EVs through their cargo of regulatory molecules, potentially NOX isotypes contributed to the modulation of polarization of monocytes to distinct macrophage phenotypes. We studied the effect of selected exogenous antioxidant/bioactive compounds isolated from industrial waste to understand their efficacy in protecting the cells against oxidative damage. The intricate equilibrium and transition between the pro-oxidant and antioxidant effects of antioxidants were also investigated.
In the present study, we investigated the polarization of monocytes to macrophages, ROS generation during the transition, and mediated protein modifications under the influence of different differentiation inducers. Morphological changes in monocytes in the presence of a differentiation inducer were studied using Confocal Laser Scanning Microscopy (CLSM), and NADPH oxidase expression was studied using Immunoblotting. The regulation of the enzyme complex was studied in both monocytes and macrophages, and the role of EVs secreted from monocytes and their effect on differentiating monocytes as messengers or antioxidant cargo was also studied. Our results showed that monocytes turn into macrophages when they encounter inflammation or differentiation and produce ROS. Polyunsaturated fatty acids (PUFA) are easily damaged by these reactive molecules and can lead to reactive intermediates or products such as aldehydes. Malondialdehyde (MDA), which is one of the final products of polyunsaturated fatty acids peroxidation led to protein modification.
Furthermore, the involvement of EVs derived from monocytes and their co-culture with non-differentiating/differentiating cells showed that EVs through their cargo of regulatory molecules, potentially NOX isotypes contributed to the modulation of polarization of monocytes to distinct macrophage phenotypes. We studied the effect of selected exogenous antioxidant/bioactive compounds isolated from industrial waste to understand their efficacy in protecting the cells against oxidative damage. The intricate equilibrium and transition between the pro-oxidant and antioxidant effects of antioxidants were also investigated.
Anotace v angličtině
In the present study, we investigated the polarization of monocytes to macrophages, ROS generation during the transition, and mediated protein modifications under the influence of different differentiation inducers. Morphological changes in monocytes in the presence of a differentiation inducer were studied using Confocal Laser Scanning Microscopy (CLSM), and NADPH oxidase expression was studied using Immunoblotting. The regulation of the enzyme complex was studied in both monocytes and macrophages, and the role of EVs secreted from monocytes and their effect on differentiating monocytes as messengers or antioxidant cargo was also studied. Our results showed that monocytes turn into macrophages when they encounter inflammation or differentiation and produce ROS. Polyunsaturated fatty acids (PUFA) are easily damaged by these reactive molecules and can lead to reactive intermediates or products such as aldehydes. Malondialdehyde (MDA), which is one of the final products of polyunsaturated fatty acids peroxidation led to protein modification.
Furthermore, the involvement of EVs derived from monocytes and their co-culture with non-differentiating/differentiating cells showed that EVs through their cargo of regulatory molecules, potentially NOX isotypes contributed to the modulation of polarization of monocytes to distinct macrophage phenotypes. We studied the effect of selected exogenous antioxidant/bioactive compounds isolated from industrial waste to understand their efficacy in protecting the cells against oxidative damage. The intricate equilibrium and transition between the pro-oxidant and antioxidant effects of antioxidants were also investigated.
The enzyme nicotinamide adenine dinucleotide phosphate (NADPH) oxidase abbreviated NOX is acknowledged for its synthesis of reactive oxygen species (ROS) and established role in host defense. However, emerging evidence underscores its escalating recognition and regulatory influence within the realm of adaptive immunity, especially the NOX2 isoform. Nevertheless, excessive production of NADPH oxidase-derived superoxide anion radical within the vasculature can be deleterious. Oxidative stress significantly contributes to heightened endothelial dysfunction and increased susceptibility to severe tissue injuries. While NOX enzymes can be identified in various cell types, the expression of distinct NOX isoforms is specific to particular cells or tissues. This specificity imparts unique physiological and pathological functions to each NOX variant. The current study is aimed at understanding the polarization of monocytes to macrophages by following specific differentiation markers, followed by evaluation of NOX expression analysis and protein modification in monocytes and macrophages. The effect of selected bioactive compounds for its efficacy and protection against oxidation of cellular proteins will also be studied.
Zásady pro vypracování
The enzyme nicotinamide adenine dinucleotide phosphate (NADPH) oxidase abbreviated NOX is acknowledged for its synthesis of reactive oxygen species (ROS) and established role in host defense. However, emerging evidence underscores its escalating recognition and regulatory influence within the realm of adaptive immunity, especially the NOX2 isoform. Nevertheless, excessive production of NADPH oxidase-derived superoxide anion radical within the vasculature can be deleterious. Oxidative stress significantly contributes to heightened endothelial dysfunction and increased susceptibility to severe tissue injuries. While NOX enzymes can be identified in various cell types, the expression of distinct NOX isoforms is specific to particular cells or tissues. This specificity imparts unique physiological and pathological functions to each NOX variant. The current study is aimed at understanding the polarization of monocytes to macrophages by following specific differentiation markers, followed by evaluation of NOX expression analysis and protein modification in monocytes and macrophages. The effect of selected bioactive compounds for its efficacy and protection against oxidation of cellular proteins will also be studied.
2. Ray, P. D., Huang, B.-W., & Tsuji, Y. (2012). Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cell Signal 24(5): 981-990. doi: 10.1016/j.cellsig.2012.01.008.
3. Qing, Xu., Swati, C., Jianhui Qu, Jonathan Jang, Moran Choe, Botond Banfi, John F Engelhardt, Zheng-Gang Liu (2016). NADPH Oxidases Are Essential for Macrophage Differentiation.The Journal of Biological Chemistry 2016 Sep 16; 291(38):20030-41. doi: 10.1074/jbc.M116.731216
4. Lambeth JD. Nox enzymes, ROS, and chronic disease: An example of antagonistic pleiotropy. Free Radical Biology and Medicine (2007) 43:332–347. doi: 10.1016/j.freeradbiomed.2007.03.027
5. Ogboo, B. C., Grabovyy, U. V., Maini, A., Scouten, S., van der Vliet, A., Mattevi, A., & Heppner, D. E. (2022). Architecture of the NADPH oxidase family of enzymes. Redox Biology, 52, 102298. doi.org/10.1016/j.redox.2022.102298
6. Maraldi, T., Angeloni, C., Prata, C., & Hrelia, S. (2021). NADPH Oxidases: Redox Regulators of Stem Cell Fate and Function. Antioxidants, 10(6), 973. doi.org/10.3390/antiox10060973
2. Ray, P. D., Huang, B.-W., & Tsuji, Y. (2012). Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cell Signal 24(5): 981-990. doi: 10.1016/j.cellsig.2012.01.008.
3. Qing, Xu., Swati, C., Jianhui Qu, Jonathan Jang, Moran Choe, Botond Banfi, John F Engelhardt, Zheng-Gang Liu (2016). NADPH Oxidases Are Essential for Macrophage Differentiation.The Journal of Biological Chemistry 2016 Sep 16; 291(38):20030-41. doi: 10.1074/jbc.M116.731216
4. Lambeth JD. Nox enzymes, ROS, and chronic disease: An example of antagonistic pleiotropy. Free Radical Biology and Medicine (2007) 43:332–347. doi: 10.1016/j.freeradbiomed.2007.03.027
5. Ogboo, B. C., Grabovyy, U. V., Maini, A., Scouten, S., van der Vliet, A., Mattevi, A., & Heppner, D. E. (2022). Architecture of the NADPH oxidase family of enzymes. Redox Biology, 52, 102298. doi.org/10.1016/j.redox.2022.102298
6. Maraldi, T., Angeloni, C., Prata, C., & Hrelia, S. (2021). NADPH Oxidases: Redox Regulators of Stem Cell Fate and Function. Antioxidants, 10(6), 973. doi.org/10.3390/antiox10060973
Přílohy volně vložené
list of biological materials and chemicals and List of publication